嗜热真菌热稳定几丁质酶基因的克隆、表达与分子改造
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摘要
几丁质(chitin)是由N-乙酰-β-D-氨基葡萄糖通过β-1,4-糖苷键连接起来的直链多聚物,是最丰富的天然高分子化合物之一,广泛分布于动物、植物及微生物体内。几丁质酶可以水解几丁质产生几丁寡糖和N-乙酰氨基葡萄糖,且广泛存在于自然界的生物中。近年来,随着几丁质酶的降解产物在医药、食品、农业、环保、造纸、化工等领域的应用越来越深入,几丁质酶作为制备几丁寡糖的重要工具越来越受到人们的重视。
国内外已筛选出多种几丁质酶生产菌,并将其分离提纯。但是,直接由真菌发酵产生几丁质酶在工业化生产中受多方面因素的限制,随着分子生物学技术的发展,人们已不满足于从原始产生生物中提取和利用几丁质酶,而是希望得到编码这种酶的基因并转入宿主进行高效低成本的表达。因此利用重组微生物实现几丁质酶的工业化生产、提高几丁质酶活性和产量,是目前较为理想的方法。
嗜热子囊菌原变种(Thermoascus aurantiacus var. aurantiacus)是一种广泛分布的,生长上限温度较高的嗜热真菌,能够在45℃～50℃的高温下很好的繁殖生长。本研究根据真菌几丁质酶的同源保守序列设计兼并引物,通过RT-PCR和Tail-PCR等技术首次从嗜热子囊菌原变种中分离得到了一种几丁质酶基因Chit-TAA,全长DNA为1826bp,在包含一个由1191个核苷酸构成的开放阅读框,编码396个氨基酸,该基因cDNA在GenBank中的登录号为GU338053。序列比对分析表明,该蛋白属于糖苷水解酶18家族几丁质酶,其中两个序列S-GGW和DG-D-DWE非常保守。这两段保守氨基酸序列与糖基水解酶(glycosylhydrolases)18家族催化活性有关。其中不变氨基酸残基Asp和Glu为几丁质酶的催化活性位点。利用基因重组的方法构建酵母分泌型表达载体,并转化到毕赤酵母中。于MD/MM平板上筛选His+Mut+表型的酵母转化子,经过PCR检测及G418抗性筛选得到多拷贝整合子,进行甲醇诱导培养。通过检测各转化子每隔24h的蛋白表达情况来筛选高效表达目的几丁质酶的工程菌株,筛选到表达酶活性最高的菌株GS-Chit-176。在甲醇的诱导下,重组蛋白得到了高效表达,第6天的表达量最高,达到0.433mg/mL,酶活力为7.25U/mL,同时对表达的几丁质酶进行了纯化,SDS-PAGE检测该蛋白的分子量为43.9kDa。该几丁质酶的最适反应温度为60℃,最适反应pH值为8.0,70℃处理30min仍有45%的相对酶活,具有较好的热稳定性。对其降解产物的研究表明,该几丁质酶降解胶体几丁质的主要产物为几丁质二糖。而且该酶对烟草赤星病菌(Alternaria alternata)、串珠镰刀菌(Fusarium moniliforme)的菌丝生长也有一定的抑制作用,对玉蜀黍平脐蠕孢(Bipolaris maydis)的孢子萌发有一定的抑制作用。
嗜热子囊菌原变种几丁质酶基因能在毕赤酵母中分泌表达,表达产物具有优良特性,这就预示了表达几丁质酶在几丁质工业和生物学领域的广阔应用前景。因此,期望能对该基因进行改造,或进一步优化表达条件,获得真正有工业应用价值的酵母工程菌株。
生物几丁质酶基因一般包含信号肽序列、催化域、富含丝苏氨酸域、几丁质结合域、C端延伸域。底物结合域的存在提高了酶在底物表面的浓度,有助于酶的催化区对底物的水解。植物和细菌几丁质酶的CHBD序列与纤维素酶的结合域(CBD)序列有较高的相似性,而且纤维素和几丁质都是通过β-1,4糖苷键连接起来的直链多聚物,结构及其相似。本研究利用融合PCR技术分别在嗜热子囊菌几丁质酶Tachit的C端和N端引入嗜热毛壳菌(Chaetomium thermophilum)纤维素酶chh1、cbh2的CBD结构域,构建表达载体转化毕赤酵母中,筛选到两株高表达量的酵母工程菌株。表达的融合蛋白Tachit-CBD1和CBD2-Tachit对胶体几丁质的活性分别提高了1.6倍和0.6,对粉状几丁质的活性提高了0.9和0.3倍。而且表达的Tachit-CBD1和CBD2-Tachit同时还具有热稳定性,最适反应温度为都为60℃,50℃条件下相对稳定。最适反应pH值都为8.0,在pH5.0-8.0之间稳定。
通过对Tachit催化区非保守氨基酸进行定点突变,构建了两个突变体Tachit-L129F和Tachit-V134I,通过比较Tachit-L129F的最适反应温度提高了10℃,而热稳定基本不变,最适反应pH值为降为5.0,在酸性条件下稳定；Tachit-V134I的性质研究结果表明,突变对该酶最适反应温度的影响变化不大。对其最适反应pH的测定表明,该突变酶表现出较好的嗜碱性,最适反应pH值为11,当pH值为12时,仍保持85%的酶活性。
Chitin as one of the most abundant natural polymeric compounds, is a linear chain polymer which is connected by N-Acetyl-β-D-glucosamine throughβ-1,4-glucosidic bond. Chitinase is widely spreading in animals, plants and microbes, and has a function of hydrolyzing chitin into chitin oligosaccharide and N-Acety glucosamine. Recently, as the wide application of degradation products from chitinase in medicine, food, agriculture, environmental protection, paper making, and chemistry, chitinase receives much attention as an important tool for chitin oligosaccharide.
Many organisms capable of producing chitinase were screened, isolated, and purified both at home and abroad. However, Chitinases from fungal fermentation have many limitations in commercial process. Following the development of Protocols in Molecular Biology, induction, production and use of chitinase from microorganism is not met to the requirements of human. The process of encoding for the enzyme and expressed in the source at a high effective and low cost way, is undertaking. So, it's ideal that commercial process of chitinase, improved the activity and enhanced the production can be achieved using a method of recombinant microorganisms.
Thermoascus aurantiacus var. aurantiacus is a wide distributed thermophilic fungus and a higher growth upper-limitation temperature, which can grow well up to 40-50 degrees. Degenerate primers based on the conserved sequences of the fungi chitinase and a chitinase gene named Chit-TAA was isolated for the first time from T. aurantiacus var. aurantiacus through RT-PCR and Tail-PCR. The full-length of DNA 1824bp contained an ORF of 1191bp encoding 396 amino acids. Accession number of the nucleotide sequence is GU338053 in the GenBank database. Sequence analysis of the deduced amino acid sequence revealed high homology with the catalytic domains of the Glycoside hydrolase family 18, contained 2 conserved motifs which were related with catalytic activity of chitinase. Among these conserve sequences, the invariant carboxylic amino acid residue Asp and Glu have been proved essential in the acid-base catalytisis of glycoside hydrolase of family 18.
Chit-TAA was expressed in Pichia pastoris GS115. Gene recombination was used to construct the expression plasmid and then the plasmid was transformed into P. pastoris GS115. After screening for His+Mut+ transformants on MD and MM plates, PCR analysis of Pichia integrants and G418 screening determined the multicopy integrants to induce by methanol. These integrants were used to analyze expression levels every 24 hours of interest protein and determine the engineering strains with high expression level, such as GS-Chit-176.After six days induction using methanol the transformant strain had the highest efficient production and the expressing level was 0.433mg/mL. The activity of expressed protein was 7.25U/mL. The molecular mass of a single band of the enzyme was estimated to be 43.9 kDa. The optimum temperature of the enzyme was 60℃and its maximal activity was at pH 8.0. The enzyme was thermostable and could still keep 45% activity at 70℃after 30 min. The product was mainly chitobiose from the hydrolyzation of colloidal chitin by chitinases. To some extent, the chitinase revealed antifungal activity against the growthing of Alternaria alternata and Fusarium moniliforme and the sporegermination of Bipolar is maydis.
The gene Chit-TAA was expressed in P. pastoris and the expression products had the activity of Chitinase. It implied the expression chtinase could be applied widely for the chitin industry and biological fields. Therefore, we hope to reconstruct the gene or optimize the expression condition to obtain the yeast engineering strains suitable for industrial application.
Chitinase comprises signal peptide coding sequence, catalysis domain (CD), serine/threorine domain, Chitinase binding domain (BD), and c-terminal extension region. The existence of BD increases the concentration of enzymes on the surface of substrate, which helps CD hydrolyze substrate. CHBD of plants and bacterium is pretty similar to'CBD of cellulases; and cellulos as well as chitin are linear chain polymer connected throughβ-1,4-glucosidic bond, which show very similar structure between each other. This study introduced CBD of Cellulase gene chhl and cbh2 from Chaetomium thermophilum to C-and N-Tachit from T. aurantiacus var. aurantiacus, separately, via fusion PCR. The gene was expressed in P. pastoris. Two high expression strains were gotten from screening. The fused protein Tachit-CBD1 and CBD2-Tachit increased activity for colloidal chitin by 1.6-and 0.6-fold, separately, and by 0.32-and 0.9-fold, separately, for powdery chitin and they also had the thermostability. The optimum temperatures were both 60℃, and their maximal activity were both at pH 8.0.They kept relative stability at 50℃and the pH in the range of 5.0 to 8.0.
Two mutants Tachit-L129F and Tachit-V134I were constructed by site-direct mutagenesis to amino acid in the CD of Tachit. The optimum temperature of Tachit-L 129 F was increased by 10℃, while the thermostable was almost the same. The optimum pH was decreased to 5.0, and kept stable under acid condition. The results of Tachit-V134I showed little effect of mutation on the optimum temperature of the enzyme. The mutant showed pretty basophilia, the maximal activity was at pH 11, and still kept 85% activity at pH 12.
国内外已筛选出多种几丁质酶生产菌,并将其分离提纯。但是,直接由真菌发酵产生几丁质酶在工业化生产中受多方面因素的限制,随着分子生物学技术的发展,人们已不满足于从原始产生生物中提取和利用几丁质酶,而是希望得到编码这种酶的基因并转入宿主进行高效低成本的表达。因此利用重组微生物实现几丁质酶的工业化生产、提高几丁质酶活性和产量,是目前较为理想的方法。
嗜热子囊菌原变种(Thermoascus aurantiacus var. aurantiacus)是一种广泛分布的,生长上限温度较高的嗜热真菌,能够在45℃～50℃的高温下很好的繁殖生长。本研究根据真菌几丁质酶的同源保守序列设计兼并引物,通过RT-PCR和Tail-PCR等技术首次从嗜热子囊菌原变种中分离得到了一种几丁质酶基因Chit-TAA,全长DNA为1826bp,在包含一个由1191个核苷酸构成的开放阅读框,编码396个氨基酸,该基因cDNA在GenBank中的登录号为GU338053。序列比对分析表明,该蛋白属于糖苷水解酶18家族几丁质酶,其中两个序列S-GGW和DG-D-DWE非常保守。这两段保守氨基酸序列与糖基水解酶(glycosylhydrolases)18家族催化活性有关。其中不变氨基酸残基Asp和Glu为几丁质酶的催化活性位点。利用基因重组的方法构建酵母分泌型表达载体,并转化到毕赤酵母中。于MD/MM平板上筛选His+Mut+表型的酵母转化子,经过PCR检测及G418抗性筛选得到多拷贝整合子,进行甲醇诱导培养。通过检测各转化子每隔24h的蛋白表达情况来筛选高效表达目的几丁质酶的工程菌株,筛选到表达酶活性最高的菌株GS-Chit-176。在甲醇的诱导下,重组蛋白得到了高效表达,第6天的表达量最高,达到0.433mg/mL,酶活力为7.25U/mL,同时对表达的几丁质酶进行了纯化,SDS-PAGE检测该蛋白的分子量为43.9kDa。该几丁质酶的最适反应温度为60℃,最适反应pH值为8.0,70℃处理30min仍有45%的相对酶活,具有较好的热稳定性。对其降解产物的研究表明,该几丁质酶降解胶体几丁质的主要产物为几丁质二糖。而且该酶对烟草赤星病菌(Alternaria alternata)、串珠镰刀菌(Fusarium moniliforme)的菌丝生长也有一定的抑制作用,对玉蜀黍平脐蠕孢(Bipolaris maydis)的孢子萌发有一定的抑制作用。
嗜热子囊菌原变种几丁质酶基因能在毕赤酵母中分泌表达,表达产物具有优良特性,这就预示了表达几丁质酶在几丁质工业和生物学领域的广阔应用前景。因此,期望能对该基因进行改造,或进一步优化表达条件,获得真正有工业应用价值的酵母工程菌株。
生物几丁质酶基因一般包含信号肽序列、催化域、富含丝苏氨酸域、几丁质结合域、C端延伸域。底物结合域的存在提高了酶在底物表面的浓度,有助于酶的催化区对底物的水解。植物和细菌几丁质酶的CHBD序列与纤维素酶的结合域(CBD)序列有较高的相似性,而且纤维素和几丁质都是通过β-1,4糖苷键连接起来的直链多聚物,结构及其相似。本研究利用融合PCR技术分别在嗜热子囊菌几丁质酶Tachit的C端和N端引入嗜热毛壳菌(Chaetomium thermophilum)纤维素酶chh1、cbh2的CBD结构域,构建表达载体转化毕赤酵母中,筛选到两株高表达量的酵母工程菌株。表达的融合蛋白Tachit-CBD1和CBD2-Tachit对胶体几丁质的活性分别提高了1.6倍和0.6,对粉状几丁质的活性提高了0.9和0.3倍。而且表达的Tachit-CBD1和CBD2-Tachit同时还具有热稳定性,最适反应温度为都为60℃,50℃条件下相对稳定。最适反应pH值都为8.0,在pH5.0-8.0之间稳定。
通过对Tachit催化区非保守氨基酸进行定点突变,构建了两个突变体Tachit-L129F和Tachit-V134I,通过比较Tachit-L129F的最适反应温度提高了10℃,而热稳定基本不变,最适反应pH值为降为5.0,在酸性条件下稳定；Tachit-V134I的性质研究结果表明,突变对该酶最适反应温度的影响变化不大。对其最适反应pH的测定表明,该突变酶表现出较好的嗜碱性,最适反应pH值为11,当pH值为12时,仍保持85%的酶活性。
Chitin as one of the most abundant natural polymeric compounds, is a linear chain polymer which is connected by N-Acetyl-β-D-glucosamine throughβ-1,4-glucosidic bond. Chitinase is widely spreading in animals, plants and microbes, and has a function of hydrolyzing chitin into chitin oligosaccharide and N-Acety glucosamine. Recently, as the wide application of degradation products from chitinase in medicine, food, agriculture, environmental protection, paper making, and chemistry, chitinase receives much attention as an important tool for chitin oligosaccharide.
Many organisms capable of producing chitinase were screened, isolated, and purified both at home and abroad. However, Chitinases from fungal fermentation have many limitations in commercial process. Following the development of Protocols in Molecular Biology, induction, production and use of chitinase from microorganism is not met to the requirements of human. The process of encoding for the enzyme and expressed in the source at a high effective and low cost way, is undertaking. So, it's ideal that commercial process of chitinase, improved the activity and enhanced the production can be achieved using a method of recombinant microorganisms.
Thermoascus aurantiacus var. aurantiacus is a wide distributed thermophilic fungus and a higher growth upper-limitation temperature, which can grow well up to 40-50 degrees. Degenerate primers based on the conserved sequences of the fungi chitinase and a chitinase gene named Chit-TAA was isolated for the first time from T. aurantiacus var. aurantiacus through RT-PCR and Tail-PCR. The full-length of DNA 1824bp contained an ORF of 1191bp encoding 396 amino acids. Accession number of the nucleotide sequence is GU338053 in the GenBank database. Sequence analysis of the deduced amino acid sequence revealed high homology with the catalytic domains of the Glycoside hydrolase family 18, contained 2 conserved motifs which were related with catalytic activity of chitinase. Among these conserve sequences, the invariant carboxylic amino acid residue Asp and Glu have been proved essential in the acid-base catalytisis of glycoside hydrolase of family 18.
Chit-TAA was expressed in Pichia pastoris GS115. Gene recombination was used to construct the expression plasmid and then the plasmid was transformed into P. pastoris GS115. After screening for His+Mut+ transformants on MD and MM plates, PCR analysis of Pichia integrants and G418 screening determined the multicopy integrants to induce by methanol. These integrants were used to analyze expression levels every 24 hours of interest protein and determine the engineering strains with high expression level, such as GS-Chit-176.After six days induction using methanol the transformant strain had the highest efficient production and the expressing level was 0.433mg/mL. The activity of expressed protein was 7.25U/mL. The molecular mass of a single band of the enzyme was estimated to be 43.9 kDa. The optimum temperature of the enzyme was 60℃and its maximal activity was at pH 8.0. The enzyme was thermostable and could still keep 45% activity at 70℃after 30 min. The product was mainly chitobiose from the hydrolyzation of colloidal chitin by chitinases. To some extent, the chitinase revealed antifungal activity against the growthing of Alternaria alternata and Fusarium moniliforme and the sporegermination of Bipolar is maydis.
The gene Chit-TAA was expressed in P. pastoris and the expression products had the activity of Chitinase. It implied the expression chtinase could be applied widely for the chitin industry and biological fields. Therefore, we hope to reconstruct the gene or optimize the expression condition to obtain the yeast engineering strains suitable for industrial application.
Chitinase comprises signal peptide coding sequence, catalysis domain (CD), serine/threorine domain, Chitinase binding domain (BD), and c-terminal extension region. The existence of BD increases the concentration of enzymes on the surface of substrate, which helps CD hydrolyze substrate. CHBD of plants and bacterium is pretty similar to'CBD of cellulases; and cellulos as well as chitin are linear chain polymer connected throughβ-1,4-glucosidic bond, which show very similar structure between each other. This study introduced CBD of Cellulase gene chhl and cbh2 from Chaetomium thermophilum to C-and N-Tachit from T. aurantiacus var. aurantiacus, separately, via fusion PCR. The gene was expressed in P. pastoris. Two high expression strains were gotten from screening. The fused protein Tachit-CBD1 and CBD2-Tachit increased activity for colloidal chitin by 1.6-and 0.6-fold, separately, and by 0.32-and 0.9-fold, separately, for powdery chitin and they also had the thermostability. The optimum temperatures were both 60℃, and their maximal activity were both at pH 8.0.They kept relative stability at 50℃and the pH in the range of 5.0 to 8.0.
Two mutants Tachit-L129F and Tachit-V134I were constructed by site-direct mutagenesis to amino acid in the CD of Tachit. The optimum temperature of Tachit-L 129 F was increased by 10℃, while the thermostable was almost the same. The optimum pH was decreased to 5.0, and kept stable under acid condition. The results of Tachit-V134I showed little effect of mutation on the optimum temperature of the enzyme. The mutant showed pretty basophilia, the maximal activity was at pH 11, and still kept 85% activity at pH 12.
引文
1.蔡勇杨,江科,闫云君.酶的定向进化策.生命的化学,2007,27：16-189.
2.陈凌.几丁质酶在抗真菌感染中的研究现状及展望.国外医学预防诊断治疗用生物制品分册,2005,28：208-210.
3.邓丽瑜,余少文,邢苗,张煜,刘刚.以增加催化结构域的基因重构方法提高里氏木霉外切型葡聚糖酶Ⅱ的活性.农业生物技术学报,2007,15：1019-1023.
4.冯俊丽,朱旭芳.微生物几丁质酶的分子生物学研究.浙江大学学报,2004,30：102-108.
5.甘中伟,杨金奎,陶南,黄静文,张克勤.刀孢轮枝菌胞外几丁质酶的基因克隆及系统发育分析.菌物学报,2008,27：368-376.
6.郭润芳,史小琴,李多川,魏青,贾英民.一种耐热几丁质酶的产生及其热稳定性的研究.微生物学通报,2008,35：481-485.郭润芳,李多川,王荣.嗜热真菌Thermometers lanuginose热稳定几丁质酶的纯化及其性质研究.微生物学报,2005,45：270-274.
8.黄彩红,杨谦,宋营琦.哈茨木霉几丁质酶基因的cDNA克隆及表达.哈尔滨工业大学学报,2008,40：1609-1612.
9.金波,蒋福升,余美荣,陈铌铍,丁志山.农杆菌介导几丁质酶及β-13-葡聚糖基因转化半夏的研究.中国中药杂志,2009,34：1765-1767.
10.孔荣,钮利喜,袁静明.易错PCR法定向进化D-海因酶的初步研究.山西大学学报(自然科学版)29：425-427.
11.蓝海燕,王长海,张丽华,刘桂珍,王岚兰,陈正华,田颖川.导入β-1,3-葡聚糖酶及几丁质酶基因的转基因可育油菜及其抗菌核病的研究.生物工程学报,2000,16：141-146.
12.蓝海燕,张丽华,王兰岚,陈正华,田颖川,王长海.表达β-1,3-葡聚糖酶及几丁质酶基因的转基因烟草及其抗真菌病的研究.遗传学报,2000,27：70-77.
13.刘立强,张维宏,刘占良,李亚宁,杨文香,刘大群.链霉菌几丁质酶基因分子检测、克隆及原核表达.植物病理学报,2005,35：141-142.
14.刘丕钢,杨谦,2005.哈茨木霉几丁质酶cDNA基因的克隆及在酿酒酵母中的表达.微生物学报,45：253-257.
15.刘丕钢,杨谦,王奇慧.哈茨木霉Chiv基因的原核表达及优化.哈尔滨工业大学学报,2008，40：1252-1255.
16.罗竞红,游自立.巴斯德毕赤酵母表达系统在外源基因表达中的研究进展.生物技术通报,2007,3：75-79.
17.彭日荷,熊爱生,李贤,范惠琴,黄晓敏,姚泉洪.苏云金芽孢杆菌crylA (c)Bt基因的合成及其在大肠杆菌中稳定表达.生物化学与生物物理学报,2001,33：219-224
18.任桂杰,王志玉,王桂亭,宋艳艳,许洪芝,温红玲.副粘病毒F1蛋白胞外非保守区对其特异性膜融合的影响.病毒学报,2005,21：274-277.
19.任桂杰,王志玉.PCR介导的定点突变与随机突变的应用.山东大学学报,2005,43：865-867.
20.宋金柱,王允,杨谦.球孢白僵菌几丁质酶基因chit1真核表达.哈尔滨工业大学学报,2009，41：98-101.
21.陶苏丹,刘佳,陈喜文,陈德富.点饱和突变技术及其在蛋白质工程中的应用.中国生物工程杂志.China Biotechnology,2007,27:82-86
22.王艳君,杨谦.角毛壳菌几丁质酶基因chi58多点突变及在毕赤酵母中的高效表达.微生物学通报,2008,35：1544-1549.
23.王颖,王俊琦,胡红焱.烟曲霉几丁质酶基因的克隆与表达.生物工程学报,2004,20：843-850.
24.魏毅,’刘开启,张传云,张世宏,李多川,潘洪玉.粉红聚端孢菌TrEn39cDNA基因的原核表达.吉林农业大学学报,2004,26：399-401.
25.吴家和,张献龙,罗晓丽,聂以春,田颖川,陈正华.转几丁质酶和葡聚糖酶基因棉花的获得及其对萎黄病的抗性.遗传学报,2004,31：183-188.
26.熊爱生,彭日荷,李贤.信号肽序列对毕赤酵母表达外源蛋白的影响.生物化学与生物物理学报,2003,32：.154-160.
27.阎瑞香,吴仲,侯建华,李明钢.重组巴斯德毕赤酵母发酵生产几丁质酶的条件优化 研究.微生物学通报,2007,34：468-471.
28.杨浩萌,王亚茹,伍宁丰,姚斌.N13D、S40E点突变提高木聚糖酶XYNB的热稳定.微生物学通报,2007,34：533-537.
29.杨雪鹏,杨寿钧,韩北忠.非解朊栖热菌HG102耐热β-糖苷酶的结构与功能研究.生物工程学报,2005,21：84-91.
30.张俊梅,杜密英.极端酶的结构特征及其在食品工业中的应用.食品工程,2007,3：17-19.
31.赵翔,霍克克,李育阳.毕赤酵母的密码子用法分析.生物工程学报,2000,16：309-311.
32.周樱,王风平,肖湘.豚鼠气单胞菌CB101中几丁质酶基因的克隆及其异源表达的全酶蛋白和酶片段的功能研究.自然科学进展,2002,12：852-856.
33.左振宇,刘大岭,胡亚冬,胡熔,姚冬生.密码子优化的重组黄曲霉毒素解毒酶(rADTZ)在毕氏酵母中组成型分泌表达的研究.中国农业科技导报,2007,9：87-94
34. Andronopoulou E., Vorgias C.E. Purification and characterization of a new hyperthermostable, allosamidin insensitive and denaturation resistant chitinase from the hyperthermophilic archaeon Thermococcus chitonophagus. Extremophiles,2003,7:43-53.
35. Ang E.X., Obbard J.P., Zhao H. Probing the molecular determinants of aniline dioxygenase substrate specificity by saturation mutagenesis. FEBS J,2007,274:928-939.
36. Baek J.M., Howell C.R., Kenerley C.M. The role of an extracellular chitinase from Trichoderma virens Gv29-8 in the biocontrol of Rhizoctonia solani. Curr Genet,1999,35: 41-50.
37. Baratto C.M., Da Silva M.V., Santi L. Expression and characterization of the 42 kDa chitinase of the biocontrol fungus Metarhizium anisopliae in Escherichia coli. Canadian Journal of Microbiology,2003,49:723-726.
38. Berger R.G. Application of genetic methods to the generation of volatile flavors. In:Hui H.Y., Katchatourians G.G. (eds.):Food Biotechnology. VCH Weinheim,1995, pp 291-295.
39. Bethel C.R., Hujer A.M., Hujer K.M. Role of Asp104 in the SHV beta Olactamase. Antimicrob Agents Chemother,2006,50:4124-4131.
40. Bidochka M.J., Tong K.I., Khachatourians G.G. Partial purification and characterization of two extracellular N-acetyl-D-glucosaminidases produced by the entomopathogenic fungus Beauveria bassiana. Can J Microbiol,1993,39:41-45.
41. Blaiseau P.L, Kunz C., Grison R., Bertheau Y., Brygoo Y. Cloning and expression of chitinase gene from the hyperparasitic fungus Aphanocladium album. Curr Genet,1992,21: 61-66.
42. Blaiseau P.L., Lafay J. Primary structure of a chitinase-encoding gene (chi1) from the filamentous fungus Aphanocladium album:similarity to bacterial chitinases. Gene,1992, 120:243-248.
43. Bogo M.R., Rota C.A., Pinto H.J.R. A chitinase encoding gene (chit1 gene) from the entomopathogen Metarhizium anisopliae:isolation and characterization of genomic and full-length cDNA. Curr Microbiol,1998,37:221-225.
44. Bolar J.P., Norelli J.L., Wong K.W., Hayes C.K., Harman G.E., Aldwinckle H.S. Expression of endochitinase from Trichoderma harzianum in transgenic apple inccreases resistance to apple scsb and reduces vigor. Phytopathology,2000,90:72-77.
45. Bonfim S.M., Cruz A.H., Jesuino R.S., Ulhoa C.J., Molinari-Madlum E.E., Soares C.M., Pereira M. Chitinase from Paracoccidioides brasiliensis:molecular cloning, structural, phylogenetic, expression and activity analysis. FEMS Immunol Med Microbiol,2006,46:269-283.
46. Cannon R.D., Niimi K.,Jenkinson H.F., Shepherd M.G. Molecular cloning and expression of the Candida albicans beta-N-acetylglucosaminidase (HEX1) gene. J Bacteriol,1994,176: 2640-2647.
47. Carsolio C., Benhamou N., Haran S., Cortes C., GutierrezA., Chet I., Herrera-Estrella A. Role of the Trichoderma harzianum endochitinase gene, ech42, in mycoparasitism. Appl Environ Microbiol,1999,65:929-935.
48. Carsolio C., Gutierrez A., Jimenez B., Van Montagu M., Herrera-Estrella A. Characterization of ech42, a Trichoderma harzianum endochitinase gene expressed during mycoparasitism. Proceedings of the National Academy of Sciences,1994,91: 10903-10907.
49. Cereghino J.L., Cregg J.M. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS microbiology Reviews,2001,24:45-66.
50. Chen K., Arnold F.H. Turning the activity of an enzyme for unusual environments: sequential random mutagenesis of subtilisin E for catalysis in dimethylformamide. Proc Natl Acad Sci USA,1993,90:5618-5622.
51. Chen S.J., Chang M.C., Cheng C.Y. Effect of induction conditions on production and excretion of Aeromonas hydrophilia chitinase by recombinant Esherichia coli. Journal of Fermentation and Bioengineering,1997,84:610-613.
52. Chet I., Baker R. Induction of suppressiweness to Rhinoclonia solani in soil. Phytopathology,1980:994-998.
53. Cohen-Kupie R., Chet I. The molecular biology of chitin digestion. Current Opinion Biotechnology,1998,9:270-277.
54. Crout D.H., Vic G. Glycosidases and glycosyl transferases in glycoside and oligosaccharide synthesis. Current Opinion in Chemical Biology,1998,2:98-111.
55. Da Silva M.V., Santi L., Staats C.C., Da Costa A.M., Colodel E.M., Driemeier D., Vainstein M.H., Schrank A. Cuticle-induced endo/exoacting chitinase CHIT30 from Metarhizium anisopliae is encoded by an ortholog of the chi3 gene. Research in Microbiology,2005, 156:382-392.
56. Daines A.M., Maltman B.A., Flitsch S.L. Synthesis and modifications of carbohydrates, using biotransformations. Current Opinion in Chemical Biology,2004,8:106-113.
57. De La Cruz J., Hidalgo-Gallego A., Lora J.M. Isolation and characterization of three chitinases from Trichoderma harzianum. Eur J Biochem,1992,206:859-867.
58. De Lisboa Marco J, Valadares-Inglis M.C, Felix C.R. Purification and characterization of an N-acetylglucosaminidase produced by a Trichoderma harzianum strain which controls Crinipellis perniciosa. Appl Microbiol Biotechnol,2004;64:70-75.
59. De Wulf P., Soetacert W., Schwengers D. Screening and mutational improvement of a D-Ribose secreting Candida pelliculosa strain. Journal of fermentation and bioengineering, 1996,82:1-7.
60. Deane E.E., Whipps J.M., Lynch J.M. The purification and characterization of a Trichoderma harzianum exochitinase. Biochimica et Biophysica Acta,1998,1383:101-110.
61. Di Giambattista R., Federici F., Petruccioli M. The chitinolytic activity of Penicillium janthinellum P9:purification, partial characterization and potential applications. J Appl Microbiol,2001,91:498-505.
62. Di Pietro A., Lorito M., Hayes C.K. Endochitinase from Gliocladium virens:isolation, characterization, and synergistic antifungal activity in combination with gliotoxin. Phytopathology,1993,83:308-313.
63. Draborg H., Christgau S., Halkier T. Secretion of an enzymatically active Trichoderma harzianum endochitinase by Saccharomyces cerevisiae. Curr Genet,1996,29:404-409.
64. Draborg H., Kauppinen S., Dalboge H., Christgau S. Molecular cloning and expression in S. cerevisiae of two exo chitinases from Trichoderma harzianum. Biochem Mol Biol Int,1995, 36:781-791.
65. Dujon B., Sherman D., Fischer G., Durrens P., Casaregola S., Lafontaine I. Genome evolution in yeasts. Nature,2004, Jul 1; 430:35-44.
66. Escott G.M., Hearn V.M., Adams D.J. Inducible chitinolytic system of Aspergillus fumigatus. Microbiology,1998,144:1575-1581.
67. Fairweather J.K., Faijes M., Driguez H. Specificity studies of Bacillus 1,3-1,4-β-glucanases and application to glycosynthase-catalyzed transglycosylation. Chem Biochem,2002,3: 866-873.
68. Fairweather J.K., Hrmova M., Rutten S.J. Synthesis of complex oligosaccharides by using a mutated(1,3)-β-D-glucan endohydrolase from barley. Chemistry,2003,9:2603-2610.
69. Fang W., Leng B., Xiao Y. Cloning of Beauveria bassiana chitinase gene Bbchitl and its application to improve fungal strain virulence. Appl Environ Microbiol,2005,71:363-370.
70. Ferrandon S., Sterzenbaeh T., Mersha F.B. A single surface tryptophan in the chitin-binding domain from Bacillus circulans chitinase A1 plays a pivotal role in binding chitin and can be modified to created an elutable affinity tag. Biochem Biophys Acta,2003,1621:31-40.
71. Flaeh J., Pilet P.E, Jolles P. What's new in chitinase research? Cellular and Molecular Life Sciences,1992,48:701-716.
72. Fort S., Boyer V., Greffe L. Highly efficient synthesis of β(1→4)-oligo-and-polysaccharides using a mutant cellulase. Journal of the American Chemical Society,2000,122:5429-5437.
73. Gabor E.M.,Janssen D.B. Increasing the synthetic performance of penicillin acylase PAS2 by structure-inspired semi-random mutagenesis. Protein Engineering Design and Selection, 2004,17:571-579.
74. Garcia I., Lora J.M., Benitez T., Benitez T., Llobell A., Pintor-Toro J.A. Cloning and characterization of a chitinase (CHIT42) cDNA from the mycoparasitic fungus Trichoderma harzianum. Curr Genet,1994,27:83-89.
75. Griswold K.E., Aiyappan N.S., Iverson B.L. The evolution of catalytic efficiency and substrate promiscuity in human theta class 1-1 glutathione transferase. Journal of Molecular Biology,2006,364:400-410.
76. Guo R.F., Li D.C. Cloning and expression of thermostable chitinase from Thermomyces lanuginosus. Acta Microbiologica Sinica,2006,46:99-103 (in Chinese).
77. Haki G.D., Rakshit S.K. Developments in industrial important thermostable enzymes:a review. Bioresour Technol,2003,89:17-34.
78. Hao J., Berry A. A thermostable variant of fructose bisphosphate aldolase constructed by directed evolution also shows increased stability in organic solvents. Protein Engineering Design Selection,2004,17:689-697.
79. Haran S., Schickler H., Oppenheim A. New components of the chitinolytic system of Trichoderma harzianum. Mycological Research,1995,99:441-446.
80. Hardt M., Laine R.A. Mutation of active site residues in the chitin-binding domain ChBD ChiAl from chitinase A1 of Bacillus circulans alters substrate specificify:use of a green fluorescent protein binding assay. Archives of Biochemistry and BioPhysics,2004,426: 286-297.
81. Harman G.E., Hayes C.K., Lorito M. Chitinolytic enzymes of Trichoderma harzianum: purification of chitobiosidase and endochitinase. Phytopathology,1993,83:313-318.
82. Hayes C.K., Klemsdal S., Lorito M. Isolation and sequence of an endochitinase-encoding gene from a cDNA library of Trichoderma harzianum. Gene,1994,138:143-148.
83. Henrissat B. Classification of chitinase modules. In:Jolles P., Muzzarelli R.A.A. (ed.) Chitin and chitinases. Birkhauser Veriag Basel,1999, pp.137-159.
84. Henrissat B., Bairoch A. New families in the classfication of glycosylhydrolases based on amino acid sequence similarities. Biochem J,1993,293:781-788.
85. Henrissat B., Bairoch A. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J,1993,293:781-788.
86. Hiroyuki K., Ikuo S., Yu L. Conjugation of RGDS peptide with CM-chitin augments the peptide mediated inhibition of tumor metastasis. Carbohydrate Polymers,1993,21: 299-307.
87. Hoell I.A., Klemsdal S.S., Vaaje-Kolstad G, Hora S.J., Eijsink V.G. Overexpression and characterization of a novel chitinase from Trichoderma atroviride strain P1. Biochim Biophys Acta,2005,1748:180-190.
88. Hrmova M., Imai T., Rutten S.J. Mutated barley (1,3)-β-D-glucan endohydrolases synthesize crystalline (1,3)-β-D-glucans. The Journal of Biological Chemistry,2002, 277(33):30102-30111.
89. Huang C.Y., Chang A.K., Nixon P.F. Site-directed mutagenesis of the ionizable groups in the active site of Zymonas mobilis pyruvate decarboxylase:effect on activity and pH dependence. Eur J Biochem,2001,268:3558-3565.
90. Ike M., Nagamatsu K., Shioya A. Purification, characterization, and gene cloning of 46 kDa chitinase (Chi46) from Trichoderma reesei PC-3-7 and its expression in Escherichia coli. Appl Micribiol Biotechnol,2006,71:294-303.
91. Ike M., Nagamatsu K., Shioya A., Nogawa M., Ogasawara W., Okada H., Morikawa Y. Purification characterization and gene cloning of 46 kDa chitinase (Chi46) from Trichoderma reesei PC-3-7 and its expression in Escherichia coli. Applied Micribiology Biotechnology,2005,71:294-303.
92. Ikeda M., Miyauchi K., Mochizuki A. Purification and characterization of chitinase from the stomach of silver croaker Pennahia argentatus. Protein Expression and Purification, 2009,65:214-222.
93. Irene G., Jose M.L., De La Cruz J., Tahia B., Antonio L., Jose A., Pintor T. Cloning and characterization of chitinase (CHIT42) cDNA from mycoparasitic fungus Trichoderma harzianum. Curr Genet,1994,27:83-89.
94. Jahn M., Chen H., Mullegger J. Thioglycosynthases:double mutant glycosidases that serve as scaffolds for thioglycoside synthesis. Chem Commun,2004,7:274-275.
95. Jahn M., Marles J., Warren R.A. Thioglycoligases:mutant glycosidases for thioglycoside synthesis.Angew Chem Int Ed Engl,2003,42:352-354.
96. Jahn M., Withers S.G. New approaches to enzymatic oligosaccharide synthesis: glycosynthases and thioglycoligases. Biocatal Biotransfor,2003,21:159-166.
97. Jakeman D.L., Withers S.G. On expanding the repertoire of glycosynthases:mutant β-galactosidases forming β-(1,6)-linkages. Canadian Journal of Chemistry,2002,80: 866-870.
98. Jaques A.K.,Fukamizo T.,Hall D.Disruption of the gene encoding the ChiB1 chitinase of Aspergillus fumigatus and characterization of a recombinant gene product. Microbiology, 2003,149:2931-2939.
99. Johannes T.W., Woodyer R.D., Zhao H.M. Directed evolution of a thermostable phosphite dehydrogenase for NAD(P) H regeneration. Appl Environ Microbiol,2005,71:5728-5734.
100.Kadam K.L., Wollweber K.L., Grouch J.C. Investigations of plasmid instability in amylase producing Bacillus subtitles using continuous culture. Biotechnology Bioeng,1987,29(7): 859-872.
101.Kang S., Park S., Lee D.G. Isolation and characterization of a chitinase cDNA from the entomopathogenic fungus, Metarhizium anisopliae. FEMS Microbiol Lett,1998,165: 267-271.
102.Kang S.C., Park S., Lee D.G. Purification and characterization of a novel chitinase from the entomopathogenic fungus, Metarhizium anisopliae. Journal of Invertebrate Pathology,1999, 73:276-281.
103.Kim D.J., Baek J.M., Uribe P., Kenerley C.M., Cook. Cloning and characterization of multiple glycosyl hydrolase genes from Trichoderma rirens. Curr Genet,2002,40: 374-384.
104.Kim S., Matsuo I., Ajisaka K., Nakajima H., Kitamoto K. Cloning and characterization of the nagA gene that encoded b-N-acetylglucosaminidase from Aspergillus nidulans and its expression in Aspergillus oryzae. Biosci Biotechnol Biochem,2002,66:2168-2175.
105.Klemsdal S.S., Clarke J.L., Hoell I.A., Eijsink V.G., Brurberg M.B. Molecular cloning, characterization, and expression studies of a novel chitinase gene (ech30) from the mycoparasite Trichoderma atroviride strain P1. FEMS Microbiol Lett,2006,256:282-289.
106.Kunkel T.A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proceedings of the National Academy of Sciences of the USA,1985,82:488-492.
107.Kunz C., Sellam O., Bertheau Y. Purification and characterization of a chitinase from the hyperparasitic fungus Aphanocladium album. Physiological and Molecular Plant Pathology, 1992a,40:117-131.
108.Kuranda M.J, Robbins P.W. Chitinase is required for cell separation during growth of Saccharomyces cerevisiae. The Journal of Biological Chemistry,1991,266:19758-19767.
109.Kuranda M.J., Robbins P.W. Cloning and heterologous expression of glycosidase genes from Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences of the USA,1987,84:2585-2589.
110.Lee Y.S., Park I.H., Yoo J.S., Chung SY., Lee Y.C., Cho Y.S., Ahn S.C., Kim C.M., Choi Y.L. Cloning, purification, and characterization of chitinase from Bacillus sp. DAU101. Bioresource Technology,2007,98:2734-2741.
111.Li A.N., Yu K., Liu H.Q., Zhang J., Li H., Li D.C. Two novel thermostable chitinase genes from thermophilic fungi:Cloning, expression and characterization. Bioresource Techonlogy, 2010,101:5546-5551.
112.Li D.C. Review of fungal chitinases. Mycopathologia,2006,161:345-360.
113.Li D.C., Zhang S.H., Liu K.Q. Purification and partial characterization of a chitinase from the mycoparasitic fungus Trichothecium roseum. The Journal of General and Applied Microbiology,2004,50:35-39.
114.Lieshout J., Faijes M., Nieto J. Hydrolase and glycosynthase activity of endo-1, 3-beta-glucanase from the thermophile Pyrococcus furiosus. Archaea,2004,1:285-292.
115.Limon M.C., Lora J.M., Garcia I., De La Cruz J., Llobell A., Benitez T., Pintor-Toro J.A. Primary structure and expression pattern of the 33kDa chitinase gene from themycoparasitic fungus Trichoderma harzianum.Curr Genet,1995,28:478-483.
116.Limon M.C., Margoles-Clark E., Benitez T. Addition of substrate-binding domains increases substrate-binding capacity and specific activity of chitins from Trichoderma harzianum. FEMS Microbiology Lett,2001,198:57-63.
117.Lin F.P., Chen H.C., Lin C.S. Site-directed mutagenesis of Asp313, Glu315, and Asp391 residues in chitinase of Aeromomas caviae. IUBMB Life,1999,48:199-204.
118.Linder M., Lindeberg G., Reinikainen T. The difference in affinity between two fungal cellulose-binding domains is dominated by a single amino-acid substitution. FEBS Lett, 1995,372:96-98.
119.Lisboa Marco J., Valadares-Inglis M.C., Felix C.R. Purification and characterization of an N-acetylglucosaminidase produced by a Trichoderma harzianum strain which controls Crinipellis perniciosa. Appl Microbiol Biotechnol,2004,64:70-75.
120.Lorito M., Woo S.L., Fernandez I.G., Colucci G., Harman G.E., Pintor-Toro J.A., Filippone E., Muccifora S., Law-rence C.B., Zoina A., Tuzun S., Scala F. Genes from mycoparasitic fungi as a source for improving plant resistance to fungal pathogens. Procceed of the National Academy of Science of the USA,1998,95:7860-7865.
121.Lorito M.K., Woo S.L., Ambrosio M.K. Snnergistic interaction between cell wall degrading enzymes and membrane affecting compounds. Molecular Plant Microbe Interactions,1996: 206-213.
122.Lu Y.M., Zen K.C., Muthukrishnan S., Kramer K J. Site-directed mutagenesis and functional analysis of active site acidic amino acid residues D142, D144 and E146 in Manduca sexta (tobacco hornworm) chitinase. Insect Biochemistry and Molecular Biology, 2002,32:1369-1382.
123.Lu Z.X., Laroche A., Huang H.C. Isolation and characterization of chitinases from Verticillium lecanii. Can J Microbiol,2005,51:1045-1055.
124.Ma Y.F., Evans D.E. Mutations of barley β-amylase that improve substrate binding affinity and thermostability. Molecular Genetics and Genomics,2001,266(3):345-352.
125.Mackenzie L.F., Wang Q.P., Warren R.A.J. Glycosynthases:mutant glycosidases for oligosaccharide synthesis. Journal of the American Chemical Society,1998,120: 5583-5584.
126.Malet C., Planas A. From P-glucanase to β-glucansynthase:glycosyl transfer to a-glycosyl fluorides catalyzed by a mutant endoglucanase lacking its catalytic nucleophile. FEBS Letters,1998,440:208-212.
127.Margolles-Clark E., Hayes C.K., Harman G.E., Penttila M.E. Improved production of Trichoderma harzianum endochitinase by expression in Trichoderma reesei. Appl Environ Microbiol,1996,62:2145-2151.
128.Mattinena M.L., Linderb M., Telemana A. Interaction between cellohexaose and cellulose binding domains from Trichoderma reesei cellulases. FEBS Letters,1997,407(3):291-296.
129.Mavromatis K., Feller G., Kokkinidis M. Cold adaptation of a psychrophilic chitinase:a mutagenesis study. Protein Engineering,2003,16:497-503.
130.Mayer C., Zechel D.L., Reid S.P. The E358S mutant of Agrobacterium sp. P-glucosidase is a greatly improved glycosynthase. FEBS Letters,2000,466:40-44.
131.McCreath K.J., Specht C.A., Robbins P.W. Molecular cloning and characterization of chitinase genes from Candida albicans.Proc Natl Acad Sci USA 1995,92:2544-2548.
132.McCreath K.J., Specht CA., Liu Y., Shepherd MG. Molecular cloning of a third chitinase gene(CHT1)from Candida albicans. Yeast 1996,12:501-504.
133.Merzendorfer H., Zimoch L. Chitin metabolism in insects:structure, function and regulation of chitin synthases and chitinases. The Journal of Experimental Biology,2003, 206:4393-4412.
134.Miyazaki K., Arnold F.H. Exploring nonnatural evolutionary pathways by saturation mutagenesis:rapid improvement of protein function. Journal Molecular Evolution,1999, 49:716-720.
135.Miyazaki K., Takenouchi M., Kondo H. Thermal stabilization of Bacillus subtilis family-11 xylanase by directed evolution. The Journal of Biological Chenmistry,2006,281: 10236-10242.
136.Moracci M., Trincone A., Perugino G. Restoration of the activity of active-site mutants of the hyperthermophilic β-glycosidase from Sulfolobus solfataricus:dependence of the mechanism on the action of external nucleophiles. Biochemistry,1998,37:17262-17270.
137.Morimoto K., Karita S., Kimura T. Cloning, sequencing, and expression of the gene encoding Clostridium paraputrificium chitinase ChiB and analysis of the function of novel cadherin-like domains and a chitin-binding domain. J Bacteriol,1997,179:7306-7314.
138.Morissette D.C., Driscoll B.T., Jabaji-Hare S. Molecular cloning, characterization, and expression of a cDNA encoding an endochitinase gene from the mycoparasite Stachybotrys elegans. Fungal Genet Biol,2003,39:276-285.
139.Mouchacca J. Thermopilic fungi in desert soils:a neglected extreme enviroment. In: Allsopp D. et al. (eds.) Microbial Diversity and Ecosystem Function, CAB International, 1995, pp.265-288.
140.Mukhopadhyay P., Roy K.B. Protein engineering of BamHI restriction endonuclease: replacement of Cys54 by Ala enhances catalytic activity. Protein Engineering,1998,11: 931-935.
140.Nashiru O., Zechel D.L., Stoll D. β-Mannosynthase:synthesis of P-mannosides with a mutant β-mannosidase. Angewandte Chemie International Edition,2001,40:417-420.
142.Nierman W.C., Pain A., Anderson M.J., Wortman J.R., Kim H.S., Arroyo J., Berriman M. Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus. Nature,2005,438:1151-1156.
143.Okuyama M., Mori H., Watanabe K. α-Glucosidase mutant catalyzes "a-glycosynthase"-type reaction. Bioscience Biotechnology Biochemistry,2002,66: 928-933.
144.Ollis D.F., Chang H.T. Batch fermentation kinetics with (unstable) recombinant cultures. Biotechnol Bioeng,1982,24:2583-2586.
145.Palcic M.M. Biocatalytic synthesis of oligosaccharides. Current Opinion Biotechnology, 1999,10:616-624.
146.PaPanikolau Y., Prag G., Tavlas G. High resolution structural analyses of mutant chitinase A complexes with substrates provide new insight into the mechanism of catalysis. Bioehemistry,2001,40:11338-11343.
147.Patil R.S., Ghormade V., Deshpande M.V. Chitinolytic enzymes:an exploration. Enzyme and Microbial Technology,2000,26:473-483.
148.Peng R.W., Guan K.M., Huang X.L. Induction and purification of two extracellular chitinases from Beauveria bassiana. Acta Microbiologica Sinica,1996,36:103-108
149.Perugino G., Trincone A., Giordano A. Activity of hyperthermophilic glycosynthases is significantly enhanced at acidic pH. Biochemistry,2003,42:8484-8493.
150.Phillip S.W.R, Charles A, Barbara B. Cloning and Expression of a Streptomyces plicatus Chitinase In E.coli. J Biol Chem,1988,263:443-447.
151.Pinto D.S., Barreto C.C., Schrank A. Purification and characterization of an extracellular chitinase from the entomopathogen Metarhizium anisopliae. Canadian Journal of Microbiology,1997,43:322-327.
152.Pishko E.J., Kirkland T.N., Cole GT. Isolation and characterization of two chitinase-endoding genes (cts1, cts2) from the fungus Coccidioides immitis. Gene,1995, 167:173-177.
153.Ramot O., Viterbo A., Friesem D. Regulation of two homodimer hexosaminidases in the mycoparasitic fungus Trichoderma asperellum by glucosamine. Current Genetics,2004,45: 205-213.
154.Revah-Moiseev S., Carroad P.A. Conversion of the enzymatic hydrolysate of shellfish waste chitin to single-cell protein. Biotechnology Bioengineering,2004,23:1067-1078.
155.Reyes F., Calatayud J., Martinez M.J. Endochitinase from Aspergillus nidulans implicated in the autolysis of its cell wall. FEMS Microbiology Letters,1989a,51:119-124.
156.Reyes F., Calatayud J., Vazquez C. β-N-acetylglucosaminidase from Aspergillus nidulans which degrades chitin oligomers during autolysis. FEMS Microbiology Letters,1989b,65: 83-88.
157.Robbins P.W., Albright C., Benfield B. Cloning and Expression of a Streptomyces plicatus Chitinase (Chitinase-63) in Escherichia coli. The Journal of Biological Chemistry,1988, 263:443-447.
158.Rodriguez J., Copa-Patino J.L., Perez-Leblic M.I. Purification and properties of a chitinase from Penicillium oxalicum autolysates. Lett Appl Microbiol,1995,20:46-49.
159.Rye C.S., Withers S.G. Glycosidase mechanisms. Current Opinion in Chemical Biology, 2000,4:573-580.
160.Sakai K., Uchiyama T., Matahira Y. Immobilization of chitinolytic enzymes and continuos production of N-acetylglucosamine with the immobilized enzymes. Journal of Fermentation and Bioengineering,1991,72:168-172.
161.Sakamoto T., Joern J.M., Arisawa A. Laboratory evolution of toluene dioxygenase to accept 4-picoline as a substrate. Applied and Environmental Microbiology,2001,67:3882-3887.
162.Sampei Z., Nagao Y., Fukazawa T. Gene cloning and deletion analysis of chitinase J fom alkalphilic Bacillus sp. stain J813. Nucleic Acids Symp Ser (Oxf),2004,48:167-168.
163.Scigelova M., Singh S., Crout D.H.G. Glycosidases—a great synthetic tool. Journal of Molecular Catalysis B:Enzymatic,1999,6:483-494.
164.Screen S.E., Hu G., St Leger R.T. Transformants of Metarhizium anisopliae sf. anisopliae over expressing chitinase from Metarhizium anisopliae sf acridum show early induction of native chitinase but are not altered in pathogenicity to Manduca sexta. Journal of Invertebrate Pathology,2001,78:260-266.
165.Sears P., Wong C.H. Toward automated synthesis of oligosaccharides and glycoproteins. Science,2001,291:2344-2350.
166.Seidl V., Huemer B., Seiboth B., Kubicek C.P. A complete survey of Trichoderma chitinases reveals three distinct subgroups of family 18 chitinases. FEBS J,2005,272: 5923-5939.
167.Shao Z., Zhao H., Arnold F.H. Random-priming in vitro recombination:an effective tool for directed evolution. Nucleic Acids Research,1998,26:681-683.
168.Sherief A.A., El-Sawah M.A., Abdel-Naby M.A. Some properties of chitinase produced by potent Aspergillus carneus strain. Appliles Microbiology and Biotechnology,1991,35: 228-230.
169.Sieber V., Martinez C.A., Arnold F.H. Libraries of hybrid proteins from distantly related sequences. Nature Biotechnology,2001,19:456-460.
170.Simpson H.D., Barras F. Funetional analysis of the carbohydrate-binding domains of Erwinia chrysanthemi Cel5 (Endoglucanase Z) and an Escherichia coli putative chitinase. J Bacteriol,1999,181:4611-4616.
171.Smith GP. Applied evolution. The progeny of sexual PCR. Nature,1994,370:324-325.
172.Song J.K., Rhee J.S. Enhancement of stability and activity of phospholipase A(1) in organic solvents by directed evolution. Biochim Biophys Acta,2001,547:370-378.
173.Song J.Z., Yang Q., Liu B.D., Chen D. Expression of the chitinase gene from Trichoderma aureoviride in Saccharomyces cerevisiae. Applied Microbiology and Biotechnology,2005, 69:39-43.
174.Souza R.F., Gomes R.C., Coelho R.R. Purification and characterization of an endochitinase produced by Colletotrichum gloeosporioides. FEMS Microbiol Lett,2003,222:45-50.
175.St Leger R.J., Charnley A.K., Cooper R.M. Cuticle-degrading enzymes of entomopathogenic fungi:Synthesis in culture on cuticle. Journal of Invertebrate Pathology, 1986,48:85-95.
176.St Leger R.J., Cooper R.M., Charnley A.K. Cuticle degrading enzymes of entomopathogenic fungi:regulation of production of chitinolytic enzymes. Journal of General Microbiology,1986,132:1509-1517.
177.St. Leger R.J., Joshi L., Bidochka M.J. Characterization and ultrastructural localization of chitinase from Metarhizium anisopliae, M. flavoviride and Beauveria bassiana during fungal invasion of host (Manduca sexta) cuticle. Appl Environ Microbiol,1996,62: 907-912.
178.Stemmer W.P.C. Molecular breeding of genes, pathways and genomes by DNA shuffling. Journal of Molecular Catalysis B:Enzymatic,2002,19-20:3-12.
179.Stemmer W.P.C. Rapid evolution of a protein in vitro by DNA shuffling. Nature,1994,370: 389-391.
180.Suginta W., Vongsuwan A., Songsiriritthigul C. Enzymatic properties of wild-type and active site mutants of chitinase A from Vibrio carchariae as revealed by HPLC-MS. FEBS J,2005,272:3376-3386.
181.Synstad B., Gaseidnes S.,Van Aalten D.M. Mutational and computational analysis of the role of conserved residues in the active site of a family 18 chitinase. Eur J Biochem,2004, 271:253-262.
182.Takaya N., Yamazaki D., Horiuchi H. Cloning and characterization of a chitinase-encoding gene (chiA) from Aspergillus nidulans, disruption of which decreases germination frequency and hyphal growth. Biosci Biotechnol Biochem,1998a,62(1):60-65.
183.Takaya N., Yamazaki D., Horiuchi H. Intracellular chitinase gene from Rhizopus oligosporus:molecular cloning and characterization. Microbiology,1998b,144: 2647-2654.
184.Takaya N., Yamazaki D., Horiuchi H. Intracellular chitinase gene from Rhizopus oligosporus:molecular cloning and characterization. Microbiology,1998b,144:2647-2654.
185.Takaya N., Ymazaki D., Horiuchi H. Cloning and characterization of a chitinase-encoding gene (chiA) from Aspergillus nidulans, disruption of which decreases germination frequency and hyphal growth. Biosci Biotechnol Biochem,1998,62:60-65.
186.Tanaka T., Fukui T., Atomi H. Characterization of an exo-β-D-glucosaminidase involved in a novel chitinolytic pathway from the hyperthermophilic archaeon Thermococcus kodakamensis KOD1.J Bacteriol,2003,185:5175-5181.
187.Thomas C.J.,Gooday G.W., King L.A. Mutagenesis of the active site coding region of the Autographa californica nucleopolyhedrovirus chiA gene. Journal of General Virology, 2000,81:1403-1411.
188.Tokura S., Itoyama K., Nishi N., Nishmura S-I., Saiki I.,Nishmura IA. Selective sulfation of chitin derivatives for biomedical functions. Journal of Macromolecular Science,1994, 31:1701-1718.
189.Trincone A., Giordano A., Perugino G. Glycosynthase-catalysed syntheses at pH below neutrality. Bioorg Med Chem Lett,2003,13:4039-4042.
190.Trincone A., Perugino G., Rossi M. A novel thermophilic glycosynthase that effects branching glycosylation. Bioorg Med Chem Lett,2000,10:365-368.
191.Tsujibo H., Miyamoto K., Tanaka K. Cloning and sequence analysis of a protease encoding gene from the marine bacterium Alteromonas sp. Stain O-7. Biosci Biotechnol Biochem, 1996,60:1284-1288.
192.Tsujibo H., Orikoshi H., Shiotani K. Characterization of chitinase C from marine bacterium, Alteromonas sp. strain 0-7, and its corresponding gene and domain structure. Applied and Environmental Microbiology,1998,64:472-478.
193.Ulhoa C.J., Peberdy J.F. Purification and some properties of the extracellular chitinase produced by Trichderma harzianum. Enzyme and Microbial Technology,1992,14: 236-240.
194.Ulhoa C.J., Peberdy J.F. Purification and some properties of the extracellular chitinase produced by Trichoderma harzianum. Enzyme and Microbial Technology,1992,14: 236-240.
195.Vaaje-Kolstad G., Houston D.R., Rao F.V. Structure of the D142N mutant of the family 18 chitinase ChiB from Serratia marcescens and its complex with allosamidin. Biochim Biophys Acta,2004,1696:103-111.
196.Valadares-Inglis M.C., Inglis P.W., Peberdy J.F. Sequence analysis of the catalytic domain of a Metarhizium anisopliae chitinase. Brazilian Journal of Genetics,1997,20:161-164.
197.Van Aalten D.M.F., Komander D., Synstad B. Structural insight into the catalytic mechanism of a family 18 exo-chitiriase. Proceedings of the National Academy of Science of the USA,2001,98:8979-8984.
198. Van den Berg M.A., Albang R., Albermann K. Genome sequencing and analysis of the filamentous fungus Penicillium chrysogenum. Nat. Biotechnol.,2008,26,1161-1168.
199.Vasseur V., Arigoni F., Andersen H. Isolation and characterizition of Aphanocladium album chitinase-overproducting mutants. Journal of General Microbiology,1990,136:2561-2567.
200.Viterbo A., Haran S., Friesem D. Antifungal activity of a novel endochitinase gene (chit36) from Trichoderma harzianum Rifai TM. FEMS Microbiology Letters,2001,200:169-174.
201.Viterbo A., Montero M., Ramot.O. Expression regulation of the endochitinase chit36 from Trichoderma asperellum (T. harzianum T-203).Curr Genet,2002,42:114-122.
202.Wada M., Hsu C.C., Franke D. Directed evolution of N-acetylneuraminic acid aldolase to catalyze enantiomeric aldol reactions. Bioorg Med Chem,2003,11:2091-2098.
203.Wang S.X., Dunphy W.G. Activation of Xenopus Chkl by mutagenesis of threonine-377. FEBS Letters,2000,487:277-281.
204.Watanabe T., Ito Y, Yamada T. The role of the C-terminal domain and type III domains of chitinase A1 from Bacillus circulans WL-12 in chitin degradation. Journal of Bacteriol, 1994,176:4465-4472.
205.Watanabe T., Kobori K., Miyashita K. Identification of glutamic acid 204 and aspartic acid 200 in chitinase A1 of Bacillus circulans WL-12 as essential residues for chitinase activity. The Journal of Biological Chemistry,1993,268:18567-18572.
206.Watanabe T., Uchida M., Kobori K..Site-directed mutagenesis of the Asp-197 and Asp-202 residues in chitinase A1 of Bacillus circulans WL-12. Biosci Biotechnol Biochem, 1994,58:2283-2285.
207.Wattanalai R., Wiwat C., Boucias D.G. Chitinase gene of the dimorphic mycopathogen, Nomuraea rileyi. J Invertebr Pathol,2004,85:54-57.
208.Wharton D.Nematode egg-shells.Parasitology,1980,81:447-463.
209.Wilson D.B., Spezio M., Irwin D. Comparison of enzymes catalyzing the hydrolysis of insoluble polysaccharides. ACS Symposium Series,1995,618:1-12.
210.Wintrode P.L., Miyazaki K., Arnold F.H. Cold adaptation of a mesophilic subtilisin-like protease by laboratory evolution. Journal of Biological Chemistry,2000,275: 31635-31640.
211.Withers S.G. Mechanisms of glycosyl transferases and hydrolases. Carbohydrate Polymers, 2001,44:325-337.
212.Xia G., Jin C., Zhou J. A novel chitinase having a unique mode of action from Aspergillus fumigatus YJ-407. Eur J Biochem,2001,268:4079-4085.
213.Yanai K., Horiuchi H., Takaya N. Preparation of protoplasts of Rhizopus niveus and their transformation with plasmid DNA. Agricultural and Biological Chemistry,1990,54: 2689-2696.
214.Yanai K., Takaya N., Kojima N. Purification of two chitinases from Rhizopus oligosporus and isolation and sequencing of the encoding genes. Journal of Bacteriology,1992,174: 7398-7406.
215.Yanai K.,Takaya N.,Kojima N. Takagi M.Purification of two chitinases from Rhizopus oligosporus and isolation and sequencing of the encoding genes.J Bacteriol, 1992,174:7398-7406.
216.Zeltins A., Schrempf H. Specific interaction of the Streptomyces chitin-binding protein CHB1 with alpha-chitin, the role of individual tryptophan residues. Eur J Biochem,1997, 246:557-564.
217.Zhang L., Meakin H., Dickinson M. Isolation of genes expressed during compatible interactions between leaf rust (Puccinia triticina) and wheat using cDNA-AFLP. Molecular Plant Pathology,2003,4:469-477.
218.Zhao H.M., Giver L., Shao Z.X. Molecular evolution by staggered extension process (StEP) in vitro recombination. Nature Biotechnology,1998,16:258-261.
219.Zimmermann C.R., Johnson S.M., Martens G.W., White A.G., Pappagianis D. Cloning and expression of the complement fixation antigen-chitinase of Coccidioides immitis. Infection and Immunity,1996,64:4967-4975.
220.Zoller M.J., Smith M. Oligonucleotide-directed mutagenesis using M13-derived vectors:an efficient and general procedure for the production of point mutations in any fragment of DNA. Nucleic Acids Research,1982,10:6487-6500. Roth S. Oligosaccharide anti-infective agents. Lancet,1996,347:1017-1021.
2.陈凌.几丁质酶在抗真菌感染中的研究现状及展望.国外医学预防诊断治疗用生物制品分册,2005,28：208-210.
3.邓丽瑜,余少文,邢苗,张煜,刘刚.以增加催化结构域的基因重构方法提高里氏木霉外切型葡聚糖酶Ⅱ的活性.农业生物技术学报,2007,15：1019-1023.
4.冯俊丽,朱旭芳.微生物几丁质酶的分子生物学研究.浙江大学学报,2004,30：102-108.
5.甘中伟,杨金奎,陶南,黄静文,张克勤.刀孢轮枝菌胞外几丁质酶的基因克隆及系统发育分析.菌物学报,2008,27：368-376.
6.郭润芳,史小琴,李多川,魏青,贾英民.一种耐热几丁质酶的产生及其热稳定性的研究.微生物学通报,2008,35：481-485.郭润芳,李多川,王荣.嗜热真菌Thermometers lanuginose热稳定几丁质酶的纯化及其性质研究.微生物学报,2005,45：270-274.
8.黄彩红,杨谦,宋营琦.哈茨木霉几丁质酶基因的cDNA克隆及表达.哈尔滨工业大学学报,2008,40：1609-1612.
9.金波,蒋福升,余美荣,陈铌铍,丁志山.农杆菌介导几丁质酶及β-13-葡聚糖基因转化半夏的研究.中国中药杂志,2009,34：1765-1767.
10.孔荣,钮利喜,袁静明.易错PCR法定向进化D-海因酶的初步研究.山西大学学报(自然科学版)29：425-427.
11.蓝海燕,王长海,张丽华,刘桂珍,王岚兰,陈正华,田颖川.导入β-1,3-葡聚糖酶及几丁质酶基因的转基因可育油菜及其抗菌核病的研究.生物工程学报,2000,16：141-146.
12.蓝海燕,张丽华,王兰岚,陈正华,田颖川,王长海.表达β-1,3-葡聚糖酶及几丁质酶基因的转基因烟草及其抗真菌病的研究.遗传学报,2000,27：70-77.
13.刘立强,张维宏,刘占良,李亚宁,杨文香,刘大群.链霉菌几丁质酶基因分子检测、克隆及原核表达.植物病理学报,2005,35：141-142.
14.刘丕钢,杨谦,2005.哈茨木霉几丁质酶cDNA基因的克隆及在酿酒酵母中的表达.微生物学报,45：253-257.
15.刘丕钢,杨谦,王奇慧.哈茨木霉Chiv基因的原核表达及优化.哈尔滨工业大学学报,2008，40：1252-1255.
16.罗竞红,游自立.巴斯德毕赤酵母表达系统在外源基因表达中的研究进展.生物技术通报,2007,3：75-79.
17.彭日荷,熊爱生,李贤,范惠琴,黄晓敏,姚泉洪.苏云金芽孢杆菌crylA (c)Bt基因的合成及其在大肠杆菌中稳定表达.生物化学与生物物理学报,2001,33：219-224
18.任桂杰,王志玉,王桂亭,宋艳艳,许洪芝,温红玲.副粘病毒F1蛋白胞外非保守区对其特异性膜融合的影响.病毒学报,2005,21：274-277.
19.任桂杰,王志玉.PCR介导的定点突变与随机突变的应用.山东大学学报,2005,43：865-867.
20.宋金柱,王允,杨谦.球孢白僵菌几丁质酶基因chit1真核表达.哈尔滨工业大学学报,2009，41：98-101.
21.陶苏丹,刘佳,陈喜文,陈德富.点饱和突变技术及其在蛋白质工程中的应用.中国生物工程杂志.China Biotechnology,2007,27:82-86
22.王艳君,杨谦.角毛壳菌几丁质酶基因chi58多点突变及在毕赤酵母中的高效表达.微生物学通报,2008,35：1544-1549.
23.王颖,王俊琦,胡红焱.烟曲霉几丁质酶基因的克隆与表达.生物工程学报,2004,20：843-850.
24.魏毅,’刘开启,张传云,张世宏,李多川,潘洪玉.粉红聚端孢菌TrEn39cDNA基因的原核表达.吉林农业大学学报,2004,26：399-401.
25.吴家和,张献龙,罗晓丽,聂以春,田颖川,陈正华.转几丁质酶和葡聚糖酶基因棉花的获得及其对萎黄病的抗性.遗传学报,2004,31：183-188.
26.熊爱生,彭日荷,李贤.信号肽序列对毕赤酵母表达外源蛋白的影响.生物化学与生物物理学报,2003,32：.154-160.
27.阎瑞香,吴仲,侯建华,李明钢.重组巴斯德毕赤酵母发酵生产几丁质酶的条件优化 研究.微生物学通报,2007,34：468-471.
28.杨浩萌,王亚茹,伍宁丰,姚斌.N13D、S40E点突变提高木聚糖酶XYNB的热稳定.微生物学通报,2007,34：533-537.
29.杨雪鹏,杨寿钧,韩北忠.非解朊栖热菌HG102耐热β-糖苷酶的结构与功能研究.生物工程学报,2005,21：84-91.
30.张俊梅,杜密英.极端酶的结构特征及其在食品工业中的应用.食品工程,2007,3：17-19.
31.赵翔,霍克克,李育阳.毕赤酵母的密码子用法分析.生物工程学报,2000,16：309-311.
32.周樱,王风平,肖湘.豚鼠气单胞菌CB101中几丁质酶基因的克隆及其异源表达的全酶蛋白和酶片段的功能研究.自然科学进展,2002,12：852-856.
33.左振宇,刘大岭,胡亚冬,胡熔,姚冬生.密码子优化的重组黄曲霉毒素解毒酶(rADTZ)在毕氏酵母中组成型分泌表达的研究.中国农业科技导报,2007,9：87-94
34. Andronopoulou E., Vorgias C.E. Purification and characterization of a new hyperthermostable, allosamidin insensitive and denaturation resistant chitinase from the hyperthermophilic archaeon Thermococcus chitonophagus. Extremophiles,2003,7:43-53.
35. Ang E.X., Obbard J.P., Zhao H. Probing the molecular determinants of aniline dioxygenase substrate specificity by saturation mutagenesis. FEBS J,2007,274:928-939.
36. Baek J.M., Howell C.R., Kenerley C.M. The role of an extracellular chitinase from Trichoderma virens Gv29-8 in the biocontrol of Rhizoctonia solani. Curr Genet,1999,35: 41-50.
37. Baratto C.M., Da Silva M.V., Santi L. Expression and characterization of the 42 kDa chitinase of the biocontrol fungus Metarhizium anisopliae in Escherichia coli. Canadian Journal of Microbiology,2003,49:723-726.
38. Berger R.G. Application of genetic methods to the generation of volatile flavors. In:Hui H.Y., Katchatourians G.G. (eds.):Food Biotechnology. VCH Weinheim,1995, pp 291-295.
39. Bethel C.R., Hujer A.M., Hujer K.M. Role of Asp104 in the SHV beta Olactamase. Antimicrob Agents Chemother,2006,50:4124-4131.
40. Bidochka M.J., Tong K.I., Khachatourians G.G. Partial purification and characterization of two extracellular N-acetyl-D-glucosaminidases produced by the entomopathogenic fungus Beauveria bassiana. Can J Microbiol,1993,39:41-45.
41. Blaiseau P.L, Kunz C., Grison R., Bertheau Y., Brygoo Y. Cloning and expression of chitinase gene from the hyperparasitic fungus Aphanocladium album. Curr Genet,1992,21: 61-66.
42. Blaiseau P.L., Lafay J. Primary structure of a chitinase-encoding gene (chi1) from the filamentous fungus Aphanocladium album:similarity to bacterial chitinases. Gene,1992, 120:243-248.
43. Bogo M.R., Rota C.A., Pinto H.J.R. A chitinase encoding gene (chit1 gene) from the entomopathogen Metarhizium anisopliae:isolation and characterization of genomic and full-length cDNA. Curr Microbiol,1998,37:221-225.
44. Bolar J.P., Norelli J.L., Wong K.W., Hayes C.K., Harman G.E., Aldwinckle H.S. Expression of endochitinase from Trichoderma harzianum in transgenic apple inccreases resistance to apple scsb and reduces vigor. Phytopathology,2000,90:72-77.
45. Bonfim S.M., Cruz A.H., Jesuino R.S., Ulhoa C.J., Molinari-Madlum E.E., Soares C.M., Pereira M. Chitinase from Paracoccidioides brasiliensis:molecular cloning, structural, phylogenetic, expression and activity analysis. FEMS Immunol Med Microbiol,2006,46:269-283.
46. Cannon R.D., Niimi K.,Jenkinson H.F., Shepherd M.G. Molecular cloning and expression of the Candida albicans beta-N-acetylglucosaminidase (HEX1) gene. J Bacteriol,1994,176: 2640-2647.
47. Carsolio C., Benhamou N., Haran S., Cortes C., GutierrezA., Chet I., Herrera-Estrella A. Role of the Trichoderma harzianum endochitinase gene, ech42, in mycoparasitism. Appl Environ Microbiol,1999,65:929-935.
48. Carsolio C., Gutierrez A., Jimenez B., Van Montagu M., Herrera-Estrella A. Characterization of ech42, a Trichoderma harzianum endochitinase gene expressed during mycoparasitism. Proceedings of the National Academy of Sciences,1994,91: 10903-10907.
49. Cereghino J.L., Cregg J.M. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS microbiology Reviews,2001,24:45-66.
50. Chen K., Arnold F.H. Turning the activity of an enzyme for unusual environments: sequential random mutagenesis of subtilisin E for catalysis in dimethylformamide. Proc Natl Acad Sci USA,1993,90:5618-5622.
51. Chen S.J., Chang M.C., Cheng C.Y. Effect of induction conditions on production and excretion of Aeromonas hydrophilia chitinase by recombinant Esherichia coli. Journal of Fermentation and Bioengineering,1997,84:610-613.
52. Chet I., Baker R. Induction of suppressiweness to Rhinoclonia solani in soil. Phytopathology,1980:994-998.
53. Cohen-Kupie R., Chet I. The molecular biology of chitin digestion. Current Opinion Biotechnology,1998,9:270-277.
54. Crout D.H., Vic G. Glycosidases and glycosyl transferases in glycoside and oligosaccharide synthesis. Current Opinion in Chemical Biology,1998,2:98-111.
55. Da Silva M.V., Santi L., Staats C.C., Da Costa A.M., Colodel E.M., Driemeier D., Vainstein M.H., Schrank A. Cuticle-induced endo/exoacting chitinase CHIT30 from Metarhizium anisopliae is encoded by an ortholog of the chi3 gene. Research in Microbiology,2005, 156:382-392.
56. Daines A.M., Maltman B.A., Flitsch S.L. Synthesis and modifications of carbohydrates, using biotransformations. Current Opinion in Chemical Biology,2004,8:106-113.
57. De La Cruz J., Hidalgo-Gallego A., Lora J.M. Isolation and characterization of three chitinases from Trichoderma harzianum. Eur J Biochem,1992,206:859-867.
58. De Lisboa Marco J, Valadares-Inglis M.C, Felix C.R. Purification and characterization of an N-acetylglucosaminidase produced by a Trichoderma harzianum strain which controls Crinipellis perniciosa. Appl Microbiol Biotechnol,2004;64:70-75.
59. De Wulf P., Soetacert W., Schwengers D. Screening and mutational improvement of a D-Ribose secreting Candida pelliculosa strain. Journal of fermentation and bioengineering, 1996,82:1-7.
60. Deane E.E., Whipps J.M., Lynch J.M. The purification and characterization of a Trichoderma harzianum exochitinase. Biochimica et Biophysica Acta,1998,1383:101-110.
61. Di Giambattista R., Federici F., Petruccioli M. The chitinolytic activity of Penicillium janthinellum P9:purification, partial characterization and potential applications. J Appl Microbiol,2001,91:498-505.
62. Di Pietro A., Lorito M., Hayes C.K. Endochitinase from Gliocladium virens:isolation, characterization, and synergistic antifungal activity in combination with gliotoxin. Phytopathology,1993,83:308-313.
63. Draborg H., Christgau S., Halkier T. Secretion of an enzymatically active Trichoderma harzianum endochitinase by Saccharomyces cerevisiae. Curr Genet,1996,29:404-409.
64. Draborg H., Kauppinen S., Dalboge H., Christgau S. Molecular cloning and expression in S. cerevisiae of two exo chitinases from Trichoderma harzianum. Biochem Mol Biol Int,1995, 36:781-791.
65. Dujon B., Sherman D., Fischer G., Durrens P., Casaregola S., Lafontaine I. Genome evolution in yeasts. Nature,2004, Jul 1; 430:35-44.
66. Escott G.M., Hearn V.M., Adams D.J. Inducible chitinolytic system of Aspergillus fumigatus. Microbiology,1998,144:1575-1581.
67. Fairweather J.K., Faijes M., Driguez H. Specificity studies of Bacillus 1,3-1,4-β-glucanases and application to glycosynthase-catalyzed transglycosylation. Chem Biochem,2002,3: 866-873.
68. Fairweather J.K., Hrmova M., Rutten S.J. Synthesis of complex oligosaccharides by using a mutated(1,3)-β-D-glucan endohydrolase from barley. Chemistry,2003,9:2603-2610.
69. Fang W., Leng B., Xiao Y. Cloning of Beauveria bassiana chitinase gene Bbchitl and its application to improve fungal strain virulence. Appl Environ Microbiol,2005,71:363-370.
70. Ferrandon S., Sterzenbaeh T., Mersha F.B. A single surface tryptophan in the chitin-binding domain from Bacillus circulans chitinase A1 plays a pivotal role in binding chitin and can be modified to created an elutable affinity tag. Biochem Biophys Acta,2003,1621:31-40.
71. Flaeh J., Pilet P.E, Jolles P. What's new in chitinase research? Cellular and Molecular Life Sciences,1992,48:701-716.
72. Fort S., Boyer V., Greffe L. Highly efficient synthesis of β(1→4)-oligo-and-polysaccharides using a mutant cellulase. Journal of the American Chemical Society,2000,122:5429-5437.
73. Gabor E.M.,Janssen D.B. Increasing the synthetic performance of penicillin acylase PAS2 by structure-inspired semi-random mutagenesis. Protein Engineering Design and Selection, 2004,17:571-579.
74. Garcia I., Lora J.M., Benitez T., Benitez T., Llobell A., Pintor-Toro J.A. Cloning and characterization of a chitinase (CHIT42) cDNA from the mycoparasitic fungus Trichoderma harzianum. Curr Genet,1994,27:83-89.
75. Griswold K.E., Aiyappan N.S., Iverson B.L. The evolution of catalytic efficiency and substrate promiscuity in human theta class 1-1 glutathione transferase. Journal of Molecular Biology,2006,364:400-410.
76. Guo R.F., Li D.C. Cloning and expression of thermostable chitinase from Thermomyces lanuginosus. Acta Microbiologica Sinica,2006,46:99-103 (in Chinese).
77. Haki G.D., Rakshit S.K. Developments in industrial important thermostable enzymes:a review. Bioresour Technol,2003,89:17-34.
78. Hao J., Berry A. A thermostable variant of fructose bisphosphate aldolase constructed by directed evolution also shows increased stability in organic solvents. Protein Engineering Design Selection,2004,17:689-697.
79. Haran S., Schickler H., Oppenheim A. New components of the chitinolytic system of Trichoderma harzianum. Mycological Research,1995,99:441-446.
80. Hardt M., Laine R.A. Mutation of active site residues in the chitin-binding domain ChBD ChiAl from chitinase A1 of Bacillus circulans alters substrate specificify:use of a green fluorescent protein binding assay. Archives of Biochemistry and BioPhysics,2004,426: 286-297.
81. Harman G.E., Hayes C.K., Lorito M. Chitinolytic enzymes of Trichoderma harzianum: purification of chitobiosidase and endochitinase. Phytopathology,1993,83:313-318.
82. Hayes C.K., Klemsdal S., Lorito M. Isolation and sequence of an endochitinase-encoding gene from a cDNA library of Trichoderma harzianum. Gene,1994,138:143-148.
83. Henrissat B. Classification of chitinase modules. In:Jolles P., Muzzarelli R.A.A. (ed.) Chitin and chitinases. Birkhauser Veriag Basel,1999, pp.137-159.
84. Henrissat B., Bairoch A. New families in the classfication of glycosylhydrolases based on amino acid sequence similarities. Biochem J,1993,293:781-788.
85. Henrissat B., Bairoch A. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J,1993,293:781-788.
86. Hiroyuki K., Ikuo S., Yu L. Conjugation of RGDS peptide with CM-chitin augments the peptide mediated inhibition of tumor metastasis. Carbohydrate Polymers,1993,21: 299-307.
87. Hoell I.A., Klemsdal S.S., Vaaje-Kolstad G, Hora S.J., Eijsink V.G. Overexpression and characterization of a novel chitinase from Trichoderma atroviride strain P1. Biochim Biophys Acta,2005,1748:180-190.
88. Hrmova M., Imai T., Rutten S.J. Mutated barley (1,3)-β-D-glucan endohydrolases synthesize crystalline (1,3)-β-D-glucans. The Journal of Biological Chemistry,2002, 277(33):30102-30111.
89. Huang C.Y., Chang A.K., Nixon P.F. Site-directed mutagenesis of the ionizable groups in the active site of Zymonas mobilis pyruvate decarboxylase:effect on activity and pH dependence. Eur J Biochem,2001,268:3558-3565.
90. Ike M., Nagamatsu K., Shioya A. Purification, characterization, and gene cloning of 46 kDa chitinase (Chi46) from Trichoderma reesei PC-3-7 and its expression in Escherichia coli. Appl Micribiol Biotechnol,2006,71:294-303.
91. Ike M., Nagamatsu K., Shioya A., Nogawa M., Ogasawara W., Okada H., Morikawa Y. Purification characterization and gene cloning of 46 kDa chitinase (Chi46) from Trichoderma reesei PC-3-7 and its expression in Escherichia coli. Applied Micribiology Biotechnology,2005,71:294-303.
92. Ikeda M., Miyauchi K., Mochizuki A. Purification and characterization of chitinase from the stomach of silver croaker Pennahia argentatus. Protein Expression and Purification, 2009,65:214-222.
93. Irene G., Jose M.L., De La Cruz J., Tahia B., Antonio L., Jose A., Pintor T. Cloning and characterization of chitinase (CHIT42) cDNA from mycoparasitic fungus Trichoderma harzianum. Curr Genet,1994,27:83-89.
94. Jahn M., Chen H., Mullegger J. Thioglycosynthases:double mutant glycosidases that serve as scaffolds for thioglycoside synthesis. Chem Commun,2004,7:274-275.
95. Jahn M., Marles J., Warren R.A. Thioglycoligases:mutant glycosidases for thioglycoside synthesis.Angew Chem Int Ed Engl,2003,42:352-354.
96. Jahn M., Withers S.G. New approaches to enzymatic oligosaccharide synthesis: glycosynthases and thioglycoligases. Biocatal Biotransfor,2003,21:159-166.
97. Jakeman D.L., Withers S.G. On expanding the repertoire of glycosynthases:mutant β-galactosidases forming β-(1,6)-linkages. Canadian Journal of Chemistry,2002,80: 866-870.
98. Jaques A.K.,Fukamizo T.,Hall D.Disruption of the gene encoding the ChiB1 chitinase of Aspergillus fumigatus and characterization of a recombinant gene product. Microbiology, 2003,149:2931-2939.
99. Johannes T.W., Woodyer R.D., Zhao H.M. Directed evolution of a thermostable phosphite dehydrogenase for NAD(P) H regeneration. Appl Environ Microbiol,2005,71:5728-5734.
100.Kadam K.L., Wollweber K.L., Grouch J.C. Investigations of plasmid instability in amylase producing Bacillus subtitles using continuous culture. Biotechnology Bioeng,1987,29(7): 859-872.
101.Kang S., Park S., Lee D.G. Isolation and characterization of a chitinase cDNA from the entomopathogenic fungus, Metarhizium anisopliae. FEMS Microbiol Lett,1998,165: 267-271.
102.Kang S.C., Park S., Lee D.G. Purification and characterization of a novel chitinase from the entomopathogenic fungus, Metarhizium anisopliae. Journal of Invertebrate Pathology,1999, 73:276-281.
103.Kim D.J., Baek J.M., Uribe P., Kenerley C.M., Cook. Cloning and characterization of multiple glycosyl hydrolase genes from Trichoderma rirens. Curr Genet,2002,40: 374-384.
104.Kim S., Matsuo I., Ajisaka K., Nakajima H., Kitamoto K. Cloning and characterization of the nagA gene that encoded b-N-acetylglucosaminidase from Aspergillus nidulans and its expression in Aspergillus oryzae. Biosci Biotechnol Biochem,2002,66:2168-2175.
105.Klemsdal S.S., Clarke J.L., Hoell I.A., Eijsink V.G., Brurberg M.B. Molecular cloning, characterization, and expression studies of a novel chitinase gene (ech30) from the mycoparasite Trichoderma atroviride strain P1. FEMS Microbiol Lett,2006,256:282-289.
106.Kunkel T.A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proceedings of the National Academy of Sciences of the USA,1985,82:488-492.
107.Kunz C., Sellam O., Bertheau Y. Purification and characterization of a chitinase from the hyperparasitic fungus Aphanocladium album. Physiological and Molecular Plant Pathology, 1992a,40:117-131.
108.Kuranda M.J, Robbins P.W. Chitinase is required for cell separation during growth of Saccharomyces cerevisiae. The Journal of Biological Chemistry,1991,266:19758-19767.
109.Kuranda M.J., Robbins P.W. Cloning and heterologous expression of glycosidase genes from Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences of the USA,1987,84:2585-2589.
110.Lee Y.S., Park I.H., Yoo J.S., Chung SY., Lee Y.C., Cho Y.S., Ahn S.C., Kim C.M., Choi Y.L. Cloning, purification, and characterization of chitinase from Bacillus sp. DAU101. Bioresource Technology,2007,98:2734-2741.
111.Li A.N., Yu K., Liu H.Q., Zhang J., Li H., Li D.C. Two novel thermostable chitinase genes from thermophilic fungi:Cloning, expression and characterization. Bioresource Techonlogy, 2010,101:5546-5551.
112.Li D.C. Review of fungal chitinases. Mycopathologia,2006,161:345-360.
113.Li D.C., Zhang S.H., Liu K.Q. Purification and partial characterization of a chitinase from the mycoparasitic fungus Trichothecium roseum. The Journal of General and Applied Microbiology,2004,50:35-39.
114.Lieshout J., Faijes M., Nieto J. Hydrolase and glycosynthase activity of endo-1, 3-beta-glucanase from the thermophile Pyrococcus furiosus. Archaea,2004,1:285-292.
115.Limon M.C., Lora J.M., Garcia I., De La Cruz J., Llobell A., Benitez T., Pintor-Toro J.A. Primary structure and expression pattern of the 33kDa chitinase gene from themycoparasitic fungus Trichoderma harzianum.Curr Genet,1995,28:478-483.
116.Limon M.C., Margoles-Clark E., Benitez T. Addition of substrate-binding domains increases substrate-binding capacity and specific activity of chitins from Trichoderma harzianum. FEMS Microbiology Lett,2001,198:57-63.
117.Lin F.P., Chen H.C., Lin C.S. Site-directed mutagenesis of Asp313, Glu315, and Asp391 residues in chitinase of Aeromomas caviae. IUBMB Life,1999,48:199-204.
118.Linder M., Lindeberg G., Reinikainen T. The difference in affinity between two fungal cellulose-binding domains is dominated by a single amino-acid substitution. FEBS Lett, 1995,372:96-98.
119.Lisboa Marco J., Valadares-Inglis M.C., Felix C.R. Purification and characterization of an N-acetylglucosaminidase produced by a Trichoderma harzianum strain which controls Crinipellis perniciosa. Appl Microbiol Biotechnol,2004,64:70-75.
120.Lorito M., Woo S.L., Fernandez I.G., Colucci G., Harman G.E., Pintor-Toro J.A., Filippone E., Muccifora S., Law-rence C.B., Zoina A., Tuzun S., Scala F. Genes from mycoparasitic fungi as a source for improving plant resistance to fungal pathogens. Procceed of the National Academy of Science of the USA,1998,95:7860-7865.
121.Lorito M.K., Woo S.L., Ambrosio M.K. Snnergistic interaction between cell wall degrading enzymes and membrane affecting compounds. Molecular Plant Microbe Interactions,1996: 206-213.
122.Lu Y.M., Zen K.C., Muthukrishnan S., Kramer K J. Site-directed mutagenesis and functional analysis of active site acidic amino acid residues D142, D144 and E146 in Manduca sexta (tobacco hornworm) chitinase. Insect Biochemistry and Molecular Biology, 2002,32:1369-1382.
123.Lu Z.X., Laroche A., Huang H.C. Isolation and characterization of chitinases from Verticillium lecanii. Can J Microbiol,2005,51:1045-1055.
124.Ma Y.F., Evans D.E. Mutations of barley β-amylase that improve substrate binding affinity and thermostability. Molecular Genetics and Genomics,2001,266(3):345-352.
125.Mackenzie L.F., Wang Q.P., Warren R.A.J. Glycosynthases:mutant glycosidases for oligosaccharide synthesis. Journal of the American Chemical Society,1998,120: 5583-5584.
126.Malet C., Planas A. From P-glucanase to β-glucansynthase:glycosyl transfer to a-glycosyl fluorides catalyzed by a mutant endoglucanase lacking its catalytic nucleophile. FEBS Letters,1998,440:208-212.
127.Margolles-Clark E., Hayes C.K., Harman G.E., Penttila M.E. Improved production of Trichoderma harzianum endochitinase by expression in Trichoderma reesei. Appl Environ Microbiol,1996,62:2145-2151.
128.Mattinena M.L., Linderb M., Telemana A. Interaction between cellohexaose and cellulose binding domains from Trichoderma reesei cellulases. FEBS Letters,1997,407(3):291-296.
129.Mavromatis K., Feller G., Kokkinidis M. Cold adaptation of a psychrophilic chitinase:a mutagenesis study. Protein Engineering,2003,16:497-503.
130.Mayer C., Zechel D.L., Reid S.P. The E358S mutant of Agrobacterium sp. P-glucosidase is a greatly improved glycosynthase. FEBS Letters,2000,466:40-44.
131.McCreath K.J., Specht C.A., Robbins P.W. Molecular cloning and characterization of chitinase genes from Candida albicans.Proc Natl Acad Sci USA 1995,92:2544-2548.
132.McCreath K.J., Specht CA., Liu Y., Shepherd MG. Molecular cloning of a third chitinase gene(CHT1)from Candida albicans. Yeast 1996,12:501-504.
133.Merzendorfer H., Zimoch L. Chitin metabolism in insects:structure, function and regulation of chitin synthases and chitinases. The Journal of Experimental Biology,2003, 206:4393-4412.
134.Miyazaki K., Arnold F.H. Exploring nonnatural evolutionary pathways by saturation mutagenesis:rapid improvement of protein function. Journal Molecular Evolution,1999, 49:716-720.
135.Miyazaki K., Takenouchi M., Kondo H. Thermal stabilization of Bacillus subtilis family-11 xylanase by directed evolution. The Journal of Biological Chenmistry,2006,281: 10236-10242.
136.Moracci M., Trincone A., Perugino G. Restoration of the activity of active-site mutants of the hyperthermophilic β-glycosidase from Sulfolobus solfataricus:dependence of the mechanism on the action of external nucleophiles. Biochemistry,1998,37:17262-17270.
137.Morimoto K., Karita S., Kimura T. Cloning, sequencing, and expression of the gene encoding Clostridium paraputrificium chitinase ChiB and analysis of the function of novel cadherin-like domains and a chitin-binding domain. J Bacteriol,1997,179:7306-7314.
138.Morissette D.C., Driscoll B.T., Jabaji-Hare S. Molecular cloning, characterization, and expression of a cDNA encoding an endochitinase gene from the mycoparasite Stachybotrys elegans. Fungal Genet Biol,2003,39:276-285.
139.Mouchacca J. Thermopilic fungi in desert soils:a neglected extreme enviroment. In: Allsopp D. et al. (eds.) Microbial Diversity and Ecosystem Function, CAB International, 1995, pp.265-288.
140.Mukhopadhyay P., Roy K.B. Protein engineering of BamHI restriction endonuclease: replacement of Cys54 by Ala enhances catalytic activity. Protein Engineering,1998,11: 931-935.
140.Nashiru O., Zechel D.L., Stoll D. β-Mannosynthase:synthesis of P-mannosides with a mutant β-mannosidase. Angewandte Chemie International Edition,2001,40:417-420.
142.Nierman W.C., Pain A., Anderson M.J., Wortman J.R., Kim H.S., Arroyo J., Berriman M. Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus. Nature,2005,438:1151-1156.
143.Okuyama M., Mori H., Watanabe K. α-Glucosidase mutant catalyzes "a-glycosynthase"-type reaction. Bioscience Biotechnology Biochemistry,2002,66: 928-933.
144.Ollis D.F., Chang H.T. Batch fermentation kinetics with (unstable) recombinant cultures. Biotechnol Bioeng,1982,24:2583-2586.
145.Palcic M.M. Biocatalytic synthesis of oligosaccharides. Current Opinion Biotechnology, 1999,10:616-624.
146.PaPanikolau Y., Prag G., Tavlas G. High resolution structural analyses of mutant chitinase A complexes with substrates provide new insight into the mechanism of catalysis. Bioehemistry,2001,40:11338-11343.
147.Patil R.S., Ghormade V., Deshpande M.V. Chitinolytic enzymes:an exploration. Enzyme and Microbial Technology,2000,26:473-483.
148.Peng R.W., Guan K.M., Huang X.L. Induction and purification of two extracellular chitinases from Beauveria bassiana. Acta Microbiologica Sinica,1996,36:103-108
149.Perugino G., Trincone A., Giordano A. Activity of hyperthermophilic glycosynthases is significantly enhanced at acidic pH. Biochemistry,2003,42:8484-8493.
150.Phillip S.W.R, Charles A, Barbara B. Cloning and Expression of a Streptomyces plicatus Chitinase In E.coli. J Biol Chem,1988,263:443-447.
151.Pinto D.S., Barreto C.C., Schrank A. Purification and characterization of an extracellular chitinase from the entomopathogen Metarhizium anisopliae. Canadian Journal of Microbiology,1997,43:322-327.
152.Pishko E.J., Kirkland T.N., Cole GT. Isolation and characterization of two chitinase-endoding genes (cts1, cts2) from the fungus Coccidioides immitis. Gene,1995, 167:173-177.
153.Ramot O., Viterbo A., Friesem D. Regulation of two homodimer hexosaminidases in the mycoparasitic fungus Trichoderma asperellum by glucosamine. Current Genetics,2004,45: 205-213.
154.Revah-Moiseev S., Carroad P.A. Conversion of the enzymatic hydrolysate of shellfish waste chitin to single-cell protein. Biotechnology Bioengineering,2004,23:1067-1078.
155.Reyes F., Calatayud J., Martinez M.J. Endochitinase from Aspergillus nidulans implicated in the autolysis of its cell wall. FEMS Microbiology Letters,1989a,51:119-124.
156.Reyes F., Calatayud J., Vazquez C. β-N-acetylglucosaminidase from Aspergillus nidulans which degrades chitin oligomers during autolysis. FEMS Microbiology Letters,1989b,65: 83-88.
157.Robbins P.W., Albright C., Benfield B. Cloning and Expression of a Streptomyces plicatus Chitinase (Chitinase-63) in Escherichia coli. The Journal of Biological Chemistry,1988, 263:443-447.
158.Rodriguez J., Copa-Patino J.L., Perez-Leblic M.I. Purification and properties of a chitinase from Penicillium oxalicum autolysates. Lett Appl Microbiol,1995,20:46-49.
159.Rye C.S., Withers S.G. Glycosidase mechanisms. Current Opinion in Chemical Biology, 2000,4:573-580.
160.Sakai K., Uchiyama T., Matahira Y. Immobilization of chitinolytic enzymes and continuos production of N-acetylglucosamine with the immobilized enzymes. Journal of Fermentation and Bioengineering,1991,72:168-172.
161.Sakamoto T., Joern J.M., Arisawa A. Laboratory evolution of toluene dioxygenase to accept 4-picoline as a substrate. Applied and Environmental Microbiology,2001,67:3882-3887.
162.Sampei Z., Nagao Y., Fukazawa T. Gene cloning and deletion analysis of chitinase J fom alkalphilic Bacillus sp. stain J813. Nucleic Acids Symp Ser (Oxf),2004,48:167-168.
163.Scigelova M., Singh S., Crout D.H.G. Glycosidases—a great synthetic tool. Journal of Molecular Catalysis B:Enzymatic,1999,6:483-494.
164.Screen S.E., Hu G., St Leger R.T. Transformants of Metarhizium anisopliae sf. anisopliae over expressing chitinase from Metarhizium anisopliae sf acridum show early induction of native chitinase but are not altered in pathogenicity to Manduca sexta. Journal of Invertebrate Pathology,2001,78:260-266.
165.Sears P., Wong C.H. Toward automated synthesis of oligosaccharides and glycoproteins. Science,2001,291:2344-2350.
166.Seidl V., Huemer B., Seiboth B., Kubicek C.P. A complete survey of Trichoderma chitinases reveals three distinct subgroups of family 18 chitinases. FEBS J,2005,272: 5923-5939.
167.Shao Z., Zhao H., Arnold F.H. Random-priming in vitro recombination:an effective tool for directed evolution. Nucleic Acids Research,1998,26:681-683.
168.Sherief A.A., El-Sawah M.A., Abdel-Naby M.A. Some properties of chitinase produced by potent Aspergillus carneus strain. Appliles Microbiology and Biotechnology,1991,35: 228-230.
169.Sieber V., Martinez C.A., Arnold F.H. Libraries of hybrid proteins from distantly related sequences. Nature Biotechnology,2001,19:456-460.
170.Simpson H.D., Barras F. Funetional analysis of the carbohydrate-binding domains of Erwinia chrysanthemi Cel5 (Endoglucanase Z) and an Escherichia coli putative chitinase. J Bacteriol,1999,181:4611-4616.
171.Smith GP. Applied evolution. The progeny of sexual PCR. Nature,1994,370:324-325.
172.Song J.K., Rhee J.S. Enhancement of stability and activity of phospholipase A(1) in organic solvents by directed evolution. Biochim Biophys Acta,2001,547:370-378.
173.Song J.Z., Yang Q., Liu B.D., Chen D. Expression of the chitinase gene from Trichoderma aureoviride in Saccharomyces cerevisiae. Applied Microbiology and Biotechnology,2005, 69:39-43.
174.Souza R.F., Gomes R.C., Coelho R.R. Purification and characterization of an endochitinase produced by Colletotrichum gloeosporioides. FEMS Microbiol Lett,2003,222:45-50.
175.St Leger R.J., Charnley A.K., Cooper R.M. Cuticle-degrading enzymes of entomopathogenic fungi:Synthesis in culture on cuticle. Journal of Invertebrate Pathology, 1986,48:85-95.
176.St Leger R.J., Cooper R.M., Charnley A.K. Cuticle degrading enzymes of entomopathogenic fungi:regulation of production of chitinolytic enzymes. Journal of General Microbiology,1986,132:1509-1517.
177.St. Leger R.J., Joshi L., Bidochka M.J. Characterization and ultrastructural localization of chitinase from Metarhizium anisopliae, M. flavoviride and Beauveria bassiana during fungal invasion of host (Manduca sexta) cuticle. Appl Environ Microbiol,1996,62: 907-912.
178.Stemmer W.P.C. Molecular breeding of genes, pathways and genomes by DNA shuffling. Journal of Molecular Catalysis B:Enzymatic,2002,19-20:3-12.
179.Stemmer W.P.C. Rapid evolution of a protein in vitro by DNA shuffling. Nature,1994,370: 389-391.
180.Suginta W., Vongsuwan A., Songsiriritthigul C. Enzymatic properties of wild-type and active site mutants of chitinase A from Vibrio carchariae as revealed by HPLC-MS. FEBS J,2005,272:3376-3386.
181.Synstad B., Gaseidnes S.,Van Aalten D.M. Mutational and computational analysis of the role of conserved residues in the active site of a family 18 chitinase. Eur J Biochem,2004, 271:253-262.
182.Takaya N., Yamazaki D., Horiuchi H. Cloning and characterization of a chitinase-encoding gene (chiA) from Aspergillus nidulans, disruption of which decreases germination frequency and hyphal growth. Biosci Biotechnol Biochem,1998a,62(1):60-65.
183.Takaya N., Yamazaki D., Horiuchi H. Intracellular chitinase gene from Rhizopus oligosporus:molecular cloning and characterization. Microbiology,1998b,144: 2647-2654.
184.Takaya N., Yamazaki D., Horiuchi H. Intracellular chitinase gene from Rhizopus oligosporus:molecular cloning and characterization. Microbiology,1998b,144:2647-2654.
185.Takaya N., Ymazaki D., Horiuchi H. Cloning and characterization of a chitinase-encoding gene (chiA) from Aspergillus nidulans, disruption of which decreases germination frequency and hyphal growth. Biosci Biotechnol Biochem,1998,62:60-65.
186.Tanaka T., Fukui T., Atomi H. Characterization of an exo-β-D-glucosaminidase involved in a novel chitinolytic pathway from the hyperthermophilic archaeon Thermococcus kodakamensis KOD1.J Bacteriol,2003,185:5175-5181.
187.Thomas C.J.,Gooday G.W., King L.A. Mutagenesis of the active site coding region of the Autographa californica nucleopolyhedrovirus chiA gene. Journal of General Virology, 2000,81:1403-1411.
188.Tokura S., Itoyama K., Nishi N., Nishmura S-I., Saiki I.,Nishmura IA. Selective sulfation of chitin derivatives for biomedical functions. Journal of Macromolecular Science,1994, 31:1701-1718.
189.Trincone A., Giordano A., Perugino G. Glycosynthase-catalysed syntheses at pH below neutrality. Bioorg Med Chem Lett,2003,13:4039-4042.
190.Trincone A., Perugino G., Rossi M. A novel thermophilic glycosynthase that effects branching glycosylation. Bioorg Med Chem Lett,2000,10:365-368.
191.Tsujibo H., Miyamoto K., Tanaka K. Cloning and sequence analysis of a protease encoding gene from the marine bacterium Alteromonas sp. Stain O-7. Biosci Biotechnol Biochem, 1996,60:1284-1288.
192.Tsujibo H., Orikoshi H., Shiotani K. Characterization of chitinase C from marine bacterium, Alteromonas sp. strain 0-7, and its corresponding gene and domain structure. Applied and Environmental Microbiology,1998,64:472-478.
193.Ulhoa C.J., Peberdy J.F. Purification and some properties of the extracellular chitinase produced by Trichderma harzianum. Enzyme and Microbial Technology,1992,14: 236-240.
194.Ulhoa C.J., Peberdy J.F. Purification and some properties of the extracellular chitinase produced by Trichoderma harzianum. Enzyme and Microbial Technology,1992,14: 236-240.
195.Vaaje-Kolstad G., Houston D.R., Rao F.V. Structure of the D142N mutant of the family 18 chitinase ChiB from Serratia marcescens and its complex with allosamidin. Biochim Biophys Acta,2004,1696:103-111.
196.Valadares-Inglis M.C., Inglis P.W., Peberdy J.F. Sequence analysis of the catalytic domain of a Metarhizium anisopliae chitinase. Brazilian Journal of Genetics,1997,20:161-164.
197.Van Aalten D.M.F., Komander D., Synstad B. Structural insight into the catalytic mechanism of a family 18 exo-chitiriase. Proceedings of the National Academy of Science of the USA,2001,98:8979-8984.
198. Van den Berg M.A., Albang R., Albermann K. Genome sequencing and analysis of the filamentous fungus Penicillium chrysogenum. Nat. Biotechnol.,2008,26,1161-1168.
199.Vasseur V., Arigoni F., Andersen H. Isolation and characterizition of Aphanocladium album chitinase-overproducting mutants. Journal of General Microbiology,1990,136:2561-2567.
200.Viterbo A., Haran S., Friesem D. Antifungal activity of a novel endochitinase gene (chit36) from Trichoderma harzianum Rifai TM. FEMS Microbiology Letters,2001,200:169-174.
201.Viterbo A., Montero M., Ramot.O. Expression regulation of the endochitinase chit36 from Trichoderma asperellum (T. harzianum T-203).Curr Genet,2002,42:114-122.
202.Wada M., Hsu C.C., Franke D. Directed evolution of N-acetylneuraminic acid aldolase to catalyze enantiomeric aldol reactions. Bioorg Med Chem,2003,11:2091-2098.
203.Wang S.X., Dunphy W.G. Activation of Xenopus Chkl by mutagenesis of threonine-377. FEBS Letters,2000,487:277-281.
204.Watanabe T., Ito Y, Yamada T. The role of the C-terminal domain and type III domains of chitinase A1 from Bacillus circulans WL-12 in chitin degradation. Journal of Bacteriol, 1994,176:4465-4472.
205.Watanabe T., Kobori K., Miyashita K. Identification of glutamic acid 204 and aspartic acid 200 in chitinase A1 of Bacillus circulans WL-12 as essential residues for chitinase activity. The Journal of Biological Chemistry,1993,268:18567-18572.
206.Watanabe T., Uchida M., Kobori K..Site-directed mutagenesis of the Asp-197 and Asp-202 residues in chitinase A1 of Bacillus circulans WL-12. Biosci Biotechnol Biochem, 1994,58:2283-2285.
207.Wattanalai R., Wiwat C., Boucias D.G. Chitinase gene of the dimorphic mycopathogen, Nomuraea rileyi. J Invertebr Pathol,2004,85:54-57.
208.Wharton D.Nematode egg-shells.Parasitology,1980,81:447-463.
209.Wilson D.B., Spezio M., Irwin D. Comparison of enzymes catalyzing the hydrolysis of insoluble polysaccharides. ACS Symposium Series,1995,618:1-12.
210.Wintrode P.L., Miyazaki K., Arnold F.H. Cold adaptation of a mesophilic subtilisin-like protease by laboratory evolution. Journal of Biological Chemistry,2000,275: 31635-31640.
211.Withers S.G. Mechanisms of glycosyl transferases and hydrolases. Carbohydrate Polymers, 2001,44:325-337.
212.Xia G., Jin C., Zhou J. A novel chitinase having a unique mode of action from Aspergillus fumigatus YJ-407. Eur J Biochem,2001,268:4079-4085.
213.Yanai K., Horiuchi H., Takaya N. Preparation of protoplasts of Rhizopus niveus and their transformation with plasmid DNA. Agricultural and Biological Chemistry,1990,54: 2689-2696.
214.Yanai K., Takaya N., Kojima N. Purification of two chitinases from Rhizopus oligosporus and isolation and sequencing of the encoding genes. Journal of Bacteriology,1992,174: 7398-7406.
215.Yanai K.,Takaya N.,Kojima N. Takagi M.Purification of two chitinases from Rhizopus oligosporus and isolation and sequencing of the encoding genes.J Bacteriol, 1992,174:7398-7406.
216.Zeltins A., Schrempf H. Specific interaction of the Streptomyces chitin-binding protein CHB1 with alpha-chitin, the role of individual tryptophan residues. Eur J Biochem,1997, 246:557-564.
217.Zhang L., Meakin H., Dickinson M. Isolation of genes expressed during compatible interactions between leaf rust (Puccinia triticina) and wheat using cDNA-AFLP. Molecular Plant Pathology,2003,4:469-477.
218.Zhao H.M., Giver L., Shao Z.X. Molecular evolution by staggered extension process (StEP) in vitro recombination. Nature Biotechnology,1998,16:258-261.
219.Zimmermann C.R., Johnson S.M., Martens G.W., White A.G., Pappagianis D. Cloning and expression of the complement fixation antigen-chitinase of Coccidioides immitis. Infection and Immunity,1996,64:4967-4975.
220.Zoller M.J., Smith M. Oligonucleotide-directed mutagenesis using M13-derived vectors:an efficient and general procedure for the production of point mutations in any fragment of DNA. Nucleic Acids Research,1982,10:6487-6500. Roth S. Oligosaccharide anti-infective agents. Lancet,1996,347:1017-1021.