耐碱性木聚糖酶高产菌株的筛选、产酶条件优化及其木聚糖酶基因的克隆
|
摘要
本实验是采用刚果红染色的方法筛选出一株耐碱性木聚糖酶高产菌株,对其性质进行研究,并得到编码木聚糖酶的基因,实现该基因在大肠杆菌中的高表达,以满足生产中的需要。
(1)根据木聚糖酶分解木聚糖底物生成的寡糖不能被刚果红染色,从而在菌落周围形成透明圈这一特性,对采集的样品进行生孢子处理,并在筛选培养基上进行初筛和复筛,最终得到一株木聚糖酶高产菌株,命名为BYG5-20N,对该菌株进行普通染色、革兰氏染色和16SrDNA的鉴定,最终确定该菌株为短小芽孢杆菌(Bacillus pumilus)。
(2)产酶菌株生长曲线和木聚糖酶酶学性质的研究:BYG5-20N菌株接种到LB培养基,6h开始进入对数生长期,24h时达到生长高峰,在36h时细菌的生长将到最低,随后出现了二次生长,细菌生长的最适pH值为7.0-8.0。
为了给菌体提供有效的营养,对培养基中的氮源和碳源进行了优化,在培养基中添加无机氮(NH4NO3)的基础上,用L_9(3~4)拉丁方设计对有机氮牛肉膏、蛋白胨、酵母提取物进行比较,最终确定产酶的最佳组合为牛肉膏1.0%、蛋白胨0.5%、酵母提取物0.5%。在即定氮源的基础上,对碳源进行优化,结果发现木聚糖的产酶效果最好,木糖其次,而葡萄糖和麦芽糖有明显的抑制作用,说明BYG5-20N所分泌的木聚糖酶为诱导型酶,并且受木聚糖和木糖的双重诱导。通过氮源和碳源的选择,木聚糖酶的活力高达150IU/ml,并未优化前提高了52.54%。
对BYG5-20N分泌木聚糖酶的酶学性质进行了研究,改变木聚糖酶作用时的温度和pH值,确定该酶的最适值及其耐受性。结果显示:该酶最适反应温度为酶反应的最适温度为50℃。在60℃时酶活仍有55.36%,证明该酶属于嗜中温酶类,不具有高温耐受性;酶反应最适pH值为7.0,在pH为8.0时剩余木聚糖酶的活力为91.1%,即使反应pH值升高到9.0和10.0时,剩余酶活力仍在75%以上,说明该菌株具有耐碱性。
通过对木聚糖酶的简单动力学分析,得出反应分解速率的两个重要参数Km=4.2297mg/ml;Vmax=0.0123mg/ml·min;对BYG5-20N的发酵液进行浓缩,并对蛋白进行定量得到该木聚糖酶的比活性为615IU/mg。
(3)编码基因的获得及蛋白表达:利用通用引物,从所筛选的菌株BYG5-20N基因组中扩增出编码木聚糖酶的基因,该基因共687个碱基,编码228个氨基酸,分子大小为25kDa,属于11木聚糖基水解酶家族。起始密码子为ATG,终止密码子为TAA,
The aim of this experiment is to sceen a high-xylanase producing bacterium, reaserch the properties of it, get the genes coding the xylanase and achieve high expression of the xylanase in E.coli,then satisfy the need of practice.
(1) It is based on the oligoses produced by the xylanases breaking down xylan substrates can’t be dye by Kongo red, so a transparent circle will be formed among the bacterium,the gathered samples,at first,are dealed with the processing of spore disposing process,then through the first sceening and the second sceening on the filtrate culture medium,at last acquire a high-xylanase producing bacterium ,naming BYG5-20N.It was confirmed BYG5-20N is Bacillus pumilus on the common dye ,Gram staining and 16S rDNA identify.
(2) Research on the growth curve of the bacillus and the properties of xylanse:BYG5-20N was inoculated in the LB culture medium,it appear the logarithm growth period at 6h after inoculation,and have the fastigium at 24h.the growth decrease into the neap at 36h,later it have a second growth.the optimum pH is 7.0-8.0.
In order to supply the effiency nutrition for the bacterium,the nitrogen source and carbon source in the culture medium were optimized.On the basis of the adding of the inorganic nitrogen(NH_4NO_3),L_9(3~4) latin design was used to compare the organic nitrogen among beef grease、peptone and yeast,it is ascertained the optium compounding is beef grease 1.0%,peptone 0.5%,yeast 0.5%. The result is xylan is the best induction for xylanase producing,xylose is next,but glucose and maltose depress the xylan deposing.It is exlained that the xylanse of BYG5-20N belong to induce enzyme.and it was double-induced by xylan and xylose. The activity of the xylanase was highly obtained as 150IU/ml,it is enhanced 52.54% than selections of nitrogen and carbon source before.
The research was also performed on the properties of xylanase,to adfirm the tempreture and pH optimum and resistant of xylanase.Both in the time of xylanase working are changed.the result is that the optimum tempreture of xylanse is 50℃,it also have 55.36% enmezy activity at 60℃,it is proved this xylanase is midtempreture-resistant.but it hasn’t the resisitant at higher tempreture.The optimum pH is 7.0, When PH is 8.0,the remain of enzyme activity is 91.1%,Even if the pH rise to 9.0 and 10.0,the remain of enzyme activity is 75%
(1)根据木聚糖酶分解木聚糖底物生成的寡糖不能被刚果红染色,从而在菌落周围形成透明圈这一特性,对采集的样品进行生孢子处理,并在筛选培养基上进行初筛和复筛,最终得到一株木聚糖酶高产菌株,命名为BYG5-20N,对该菌株进行普通染色、革兰氏染色和16SrDNA的鉴定,最终确定该菌株为短小芽孢杆菌(Bacillus pumilus)。
(2)产酶菌株生长曲线和木聚糖酶酶学性质的研究:BYG5-20N菌株接种到LB培养基,6h开始进入对数生长期,24h时达到生长高峰,在36h时细菌的生长将到最低,随后出现了二次生长,细菌生长的最适pH值为7.0-8.0。
为了给菌体提供有效的营养,对培养基中的氮源和碳源进行了优化,在培养基中添加无机氮(NH4NO3)的基础上,用L_9(3~4)拉丁方设计对有机氮牛肉膏、蛋白胨、酵母提取物进行比较,最终确定产酶的最佳组合为牛肉膏1.0%、蛋白胨0.5%、酵母提取物0.5%。在即定氮源的基础上,对碳源进行优化,结果发现木聚糖的产酶效果最好,木糖其次,而葡萄糖和麦芽糖有明显的抑制作用,说明BYG5-20N所分泌的木聚糖酶为诱导型酶,并且受木聚糖和木糖的双重诱导。通过氮源和碳源的选择,木聚糖酶的活力高达150IU/ml,并未优化前提高了52.54%。
对BYG5-20N分泌木聚糖酶的酶学性质进行了研究,改变木聚糖酶作用时的温度和pH值,确定该酶的最适值及其耐受性。结果显示:该酶最适反应温度为酶反应的最适温度为50℃。在60℃时酶活仍有55.36%,证明该酶属于嗜中温酶类,不具有高温耐受性;酶反应最适pH值为7.0,在pH为8.0时剩余木聚糖酶的活力为91.1%,即使反应pH值升高到9.0和10.0时,剩余酶活力仍在75%以上,说明该菌株具有耐碱性。
通过对木聚糖酶的简单动力学分析,得出反应分解速率的两个重要参数Km=4.2297mg/ml;Vmax=0.0123mg/ml·min;对BYG5-20N的发酵液进行浓缩,并对蛋白进行定量得到该木聚糖酶的比活性为615IU/mg。
(3)编码基因的获得及蛋白表达:利用通用引物,从所筛选的菌株BYG5-20N基因组中扩增出编码木聚糖酶的基因,该基因共687个碱基,编码228个氨基酸,分子大小为25kDa,属于11木聚糖基水解酶家族。起始密码子为ATG,终止密码子为TAA,
The aim of this experiment is to sceen a high-xylanase producing bacterium, reaserch the properties of it, get the genes coding the xylanase and achieve high expression of the xylanase in E.coli,then satisfy the need of practice.
(1) It is based on the oligoses produced by the xylanases breaking down xylan substrates can’t be dye by Kongo red, so a transparent circle will be formed among the bacterium,the gathered samples,at first,are dealed with the processing of spore disposing process,then through the first sceening and the second sceening on the filtrate culture medium,at last acquire a high-xylanase producing bacterium ,naming BYG5-20N.It was confirmed BYG5-20N is Bacillus pumilus on the common dye ,Gram staining and 16S rDNA identify.
(2) Research on the growth curve of the bacillus and the properties of xylanse:BYG5-20N was inoculated in the LB culture medium,it appear the logarithm growth period at 6h after inoculation,and have the fastigium at 24h.the growth decrease into the neap at 36h,later it have a second growth.the optimum pH is 7.0-8.0.
In order to supply the effiency nutrition for the bacterium,the nitrogen source and carbon source in the culture medium were optimized.On the basis of the adding of the inorganic nitrogen(NH_4NO_3),L_9(3~4) latin design was used to compare the organic nitrogen among beef grease、peptone and yeast,it is ascertained the optium compounding is beef grease 1.0%,peptone 0.5%,yeast 0.5%. The result is xylan is the best induction for xylanase producing,xylose is next,but glucose and maltose depress the xylan deposing.It is exlained that the xylanse of BYG5-20N belong to induce enzyme.and it was double-induced by xylan and xylose. The activity of the xylanase was highly obtained as 150IU/ml,it is enhanced 52.54% than selections of nitrogen and carbon source before.
The research was also performed on the properties of xylanase,to adfirm the tempreture and pH optimum and resistant of xylanase.Both in the time of xylanase working are changed.the result is that the optimum tempreture of xylanse is 50℃,it also have 55.36% enmezy activity at 60℃,it is proved this xylanase is midtempreture-resistant.but it hasn’t the resisitant at higher tempreture.The optimum pH is 7.0, When PH is 8.0,the remain of enzyme activity is 91.1%,Even if the pH rise to 9.0 and 10.0,the remain of enzyme activity is 75%
引文
[1]沈萍等.微生物学[M].高等教育出版社,2004年第1版.
[2]J.萨姆布鲁克,D.W.拉塞尔著,黄培堂等译.分子克隆实验指南(第三版)[M],科学出版社,2002
[3]郭勇等,酶工程[M].中国轻工业出版社,1997.
[4]周晨妍,邬敏辰.木聚糖酶的酶学特性与分子生物学[J].生物技术,2005,15(3):89-92.
[5]刘瑞田,曲音波,杨建云.木聚糖酶分子结构研究进展[J].纤维素科学与技术,1997,5(3):1-6.
[6]张红莲,姚斌,范云六.木聚糖酶的分子生物学及其应用[J].生物技术通报,2002,3:23-30.
[7]韩晓芳,郑连爽.产木聚糖酶嗜碱细菌的筛选及产酶条件研究[J].环境污染智力技术与设备,2002,3(11):25-27.
[8] 曾莹 , 熊志刚 , 李彦 . 啤酒糟产木聚糖酶的菌种筛选及其产酶条件研究 [J]. 粮油食品科技,2005,13(2):4-6.
[9]Barbara Decelle,Adrian Tsang,Reginald K.Storms.Cloning,functional expression and characterization of three PHanernchaete chrysosporium endo-1,4-β-xylanase [J].Current Genet,2004,46:166-175.
[10]Rifaat Rawashdeh,Ismail Saadoun,Amjad Mahasneh.Effect of cultural conditions on xylanase production by Streptomyces so.(strain ib 24D)and its potential to utilize tomato pomace[J].Arican Journal of Biotechnology,2005,4(3):251-255.
[11]H.Hayashi,M.Takehara,T.Hattori,et al.Nucleotide sequences of two contiguous and highlyhomologous xylanase genes xynA and xynB and characterization of XynA from Clostridium thermocellum.[J].Appl Microbiol Biotechnol,1999,(51):348-357.
[12]Satoshi Nakamura.Structure and function of a multidomain alkaline xylanase from alkalipHilic Bacillus sp.strain 41M-1[J].Catalysis Surveys from Asia, 2003,7(2):157-164.
[13]Suren Singh,Andreas M.Madlala,Bernard A.Prior.Thermomyces lanuginosus: properties of strains and their hemicellulase[J].FEMS Microbiology Reviews,2003,27:3-16.
[14]Badal C.Saha.Hemicellulose bioconversion[J].J Ind MicrobioI Biotechnol, 2003,30:279-291.
[15]Neeta Kulkarnnni,Abhay Shendye,Mala Rao.Molecular and biotechnological aspects of xylanases [J]. FEMS Microbiology Reviews,1999,23:411-456.
[16]Licia Lama,Valeria Calandrelli,Agata Gambacorta,et al.Purification and characterization of thermostab -le xylanase shan β-xylosidase by the thermopHilic bacterium Bacillus thermantarcticus[J].Research in Microbiology, 2004,155:283-289.
[17]Q.K.Beg,M.Kapoor,L.Mahajan,et al.Microbial xylanases and their industrial applications:a review [J]. Appl Microbiol Biotechnol,2001,56:326-338.
[18]Priya Chaudhary,D.N.Deobagkar.Characterization of Cloned Endoxylanase from Cellulomonas sp.NCI -M2353 Expressed in Escherichia coli[J].Current Microboilogy, 1997;34:273-279.
[19]P.P.Dwivedi,M.D.Gibbs,D.J.Saul,et al.Cloning,sequencing and over expressing in Escherichia coli of a xylanase gene,xynA from the thermopHilic bacterium Rt8B.4 genus Caldicelluslosiruptor[J].Appl Microbiol Biotechnol, 1996,45:86-93.
[20]Oscar Gallardo,Pilar Diaz,F.I.Javier Pastor.Cloning and Characterization of Xylanase A from the strain Bacillus sp.BP-7:Comparision with Alkaline pI-Low Molecular Weight Xylanases of Family 11[J]. Cur -rent Microbiology,2004,48:276-279.
[21]O.Gallardo,P.Diaz,F.I.J.Pastor.Charaterization of a Paenibacillus cell-associa ted xylanase with highactivity on aryl-xylosides:a mew subclass of family 10 xylanases[J].Appl Microbiol Biotechnol , 2003,61: 226-233.
[22]Mursheda K.Ali,Frederick B.RudolpH,George N.Bennett.Characterization of thermostable Xyn10A enzyme form mesopHilic Clostridiun acetobutylicum ATCC824[J].Ind Microbiol Biotechnol,2005,32: 12-18.
[23]杨翔华,洪新.产木聚糖酶菌株的筛选及其产酶特性[J].辽宁石油化工大学学报,2004,24(4):32-35.
[24]Mursheda K.Ali,Frederick B.RudolpH,George N.Bennett.Thermostable xulanase 10B from Clostridiun acetobutylicum ATCC824[J].J Ind Microbiol Biotechnol,2004,31: 229-234.
[25]Hyeon Jin Sun,Shigeki Yoshida,Yasuyuki Kawabata,et al.Separatuin of two functiuonal domains of the family F/10 β-xylanase from Steptomyces olicaceociridis E-86 limited oeoteolysis with papain and some of their properties[J].Biotechnology Letter,2002,24:595-601.
[26]Hyun S Jun,Jong K.Ha,Laercio M.Malburg et al.Charateristics of a cluster of xylanase genes in Fibrobacter succinogenes S85[J].Canadian Journal of Microbiology,2003,49 (3):171-180.
[27]M.Bztaillon,AP.Nunes Carsinali,F.Duchiron.Production of xylanases from a newly isolated alkalopHiic thermopHilic Bacillus sp[J].Biotechbnology Letter,1998,20(11): 1067-1071.
[28]Laurie JI,Clarke JH,Ciruela A,et al.The Nodb domain of a multidomain xylanases from Cellulomonas fimi deacetylates acetlxylan.FEMS Letter,1997,148:261-264.
[29]Daniel D.Morris,Moreland N.Gibbs,Michelle Ford,et al. Family 10 and 11 xylanase genes from Caldicellulosiruptor sp.strain RT69B.1 [J].ExtermopHiles.1999,3:103-111.
[30]A .Shlacher,K Holzmann,M.Hayn,et al.Cloning and characterization of the gene for the thermostable xylanase XynA from Thermomyced lanuginosus[J].Journal of Biotechnology.1994,49:211-218.
[31]Hania Kebir,Claude Dupont,Rolf Morosoli.Increased xylanase production in Streptomyces lividans after replacement of the signal peptide:dependence on box and inverted repeat sequence[J].Biochimica et BiopHysica Acta,2000,1491:177-184.
[32]Asbah F.Qureshy,L A.Khan,S.Khanna.Expression of Bacillus circulans Teri-42 xylanase gene in Bacillus subtilis[J].Enzyme and Microbial Technology.2000,27:227-233.
[33]刘明启,孙建义,许英蕾.木聚糖酶分子进化的研究进展[J].中国生物工程杂志,2003,23(10):62-66.
[34]丁少军,J.A.BUSWELL.利用真菌纤维素结合域(CBD)保守性序列进行草菇木聚糖酶cDNA 的克隆[J].中国生物化学与分子生物学报,2004,20(3):408-412.
[35]T.Kmiura,T.Mizutani,T.Tanaka,et,al.Molecular breeding of transgenic rice expressing a xylanase doma -in of the xynA gene from Clostridium thermmocellum[J].Appl Microbiol Biotechnol, 2003,(62): 374- 379.
[36]G.P.Xue.J.S.Johnson,D.J.Smyth et al.Temperature-regulated expression of the tac/lacI system for overproduction of a fungal xylanase in Escherichia coli[J].Appl Microbiol Biotechnol.1996, 45:120-126.
[37]H.Kim,H.Jong,M.Y.Pack..Molecular characterization of xynX,a gene encoding a multidomain xylanase with a thermostabilizing domain form Clostridium thermocellum[J] .Appl Microbiol Biotechnol (2000) 54:521-527.
[38]Sirin S.I.Adham,Pilar Honrubia,Matgarita Diaz,et al.Expression of the genes coding for the xylanase Xys1 and the cellulose Cel1 from the straw-decompising Streptomyces halstedii JM8 cloned into the amino acid peoducet Brevibacterium lactofermentum ATCC13869[J].Arch Microbiol.201,177:91-97.
[39]Kaveh Emami,Ethan Hack.Conservation of XYN11A and XYN11B Xylanase Genes in Bipolaris sorghicola,Cochliobolus sativus, Cochliobolus heterostropHus,and Cochlo\iobilus specifer[J].Current Microbiology,2005:45:303-306.
[40]M.Uma Maheswari,T.S.Chandra.Production and potential application of a xylanas from a new strain of Streptomyces cuspidosporus[J].World Journal of Microbiology & Biotechnology,2000,16:257-263.
[41]Chartchai Khanongnuch,Saidamim Lumyong,Toshihiko Ooi,et al.A non-cellulase producing strain of Bacillus subtilis and its potential use in pulp biobleaching.Biotechnology Letter,1999,21:61-63.
[42]F.Fernadez,R.Sharma,M.Hinton,et al.Diet influences the colonidation of Campylobacter jejuni and distribution of mucin carbohydrates in the chick intestinal tract [J].Cell. Mol.Life. Science, 2000, 57: 1793-1801.
[43]J.H.Clarke,J.E.Rixon,A.Ciruela,et al.Family-10 and Family-11 xylanase differ in their capacity to enha -nce the bleachability of hardwoof and softwood paper pulps[J].Appl Microbiol Biotechnol , 1997, 48: 177-183.
[44]Chandralata Raghukumar,Usha Muraleedhara n,V.R.Gaud,et al.Xylanase of marine fungi of potential use for biobleaching of paper pulp[J].J Ind Microbiol Biotechnol,2004,31:433-441.
[45]何为,詹怀宇.木聚糖酶酶活影响因素的研究[J].中国造纸学报,2009,19(1):163-166.
[46]苏纯阳,程学慧,刘涛等.新一代的饲用酶制剂-细菌性木聚糖酶[J].饲料工业,2004,12:41-43.
[47]王金全,蔡辉益,张姝等.木聚糖酶对0-3周龄肉仔鸡肠道食糜黏度和生产性能的影响[J].中国家禽学报,2004,8(1):122-125.
[48]刘明启,孙建义,郭爽.木聚糖酶嗜酸性、热稳定性研究进展[J].中国饲料,2004,13:21-22.
[49]吴建忠,李绿雄.木聚糖酶在畜禽营养中的研究及应用[J].农业与技术,2005,25(2):73-77.
[50]方俊.木聚糖酶在猪饲料中的研究与应用[J].中国畜牧兽医,2003,30(3):20-22.
[51]王修启,林东康.皖麦38添加木聚糖酶对鸡表观代谢能值和氨基酸表观消化率的影响[J].华北农学报,2003,18(1):116-118.
[52]王海英,呙于明,袁建敏.小麦日粮中添加木聚糖酶对肉仔鸡生产性能和养分消化率的影响[J].粮食与饲料工业,2003,12:53-55.
[53]夏枚生.高麸日粮中添加复合酶制剂对仔猪血液中几种激素水平的影响[J].浙江农业科学,2000,6:300-303.
[54]王振来,钟艳玲,路广计等.木聚糖酶、β-葡聚糖酶和纤维素酶促进仔猪生长机理探讨[J].中国饲料,2004,9:20-23.
[55]Yunben B.Wu,Velmurugu Ravindran. Influence of whole wheat inclusion and xylanase supplementati- on on the performance, digestive tract measurements and carcass characteristics of broiler chickens[J]. Animal Feed Science and Technology,2004,116:129–139.
[56]Ravindran, V., Cabahug, S., Ravindran, et al. Influence of microbial pHytase on apparent ileal amino acid digestibility in feedstuffs for broilers[J]. Poult. Sci. 1999,78:699–706.
[57]F.Fernadez,R.Sharma,M.Hinton,et al.Diet influences the coloniation of Campylobacter jejuni and dis -tribution of mucin carbohydrates in the chick intestinal tract[J] Cell Mol.Life.Science, 2000,57: 1793- 1801.
[58]Ashita Dhillon,J.KGupta,B.M.Jauhari,et al.A celluase–poor thermstable,alkalitolerant xylanase produ -ced by Bacillus circulans AB16 grown on rice straw ans its application in biobleaching of eucalyptus pulp[J].Bioresource Technology.2000,(73):273-277.
[59]Chandralata Raghukumar,Usha Muraleedharan V.R,Gaud R.Mishra. Xylanase of marine fungi of potential use for biobleaching of paper pulp[J].J Ind Microbiol Biotechnol,2004,31: 433-441.
[60]Sunita Chauhan,B.Choudhury,S.N. Singh,et al.Application of xylanase enzyme of Bacillus coagulans as a prebleaching agent on non-woody pulps[J].Process Biochemistry, 2006,41 226–231.
[60]Chartchai Khanongnuch,Saidamim Lumyong,Toshihiko Ooi,et al.A non-cellulase producing strain of Bacillus subtilis and its potential use in pulp biobleaching.Biotechnology Letter,1999,21:61-63.
[61]M. Blanca Roncero,Antonio L. Torres,Jos_e F. Colom,et al.The effect of xylanase on lignocellulosic components during the bleaching of wood pulps[J].Bioresource Technology,2005, 96:21–30.
[62]J.H.Clarke,J.E.Rixon,A.Ciruela,H.J.Gilbert,G.P.Hazlewood.Family-10 and Family-11 xylanase differ in their capacity to enhance the bleachability of hardwoof and softwood paper pulps[J].Appl Microbiol Biotechnol ,1997,48:177-183.
[63]Xiaoping Yuan,Jing Wang,Huiyuan Yao.Production of feruloyl oligosaccharides from wheat bran insoluble dietary fibre by xylanases from Bacillus subtilis[J].Food Chemistry,2006,95:484–492.
[64]I.Trogh,C.M. Courtin,A.A.M.Andersson,P.A man,et al. Delcour.The combined use of hull-less barley flour and xylanase as a strategy for wheat/hull-less barley flour breads with increased arabinoxylan and (1/3,1/4)-b-D-glucan levels[J].Journal of Cereal Science, 2004,40:257–267.
[65]Tony Collins,Anne Hoyoux,Agne`s Dutron,et al.Use of glycoside hydrolase family 8 xylanases in baking[J].Journal of Cereal Science,2006,43:79–84.
[66]沈萍,范容秀,李广武主编.微生物实验[M].高等教育出版社(第三版),1999,6.
[67]Cristina Lopez,Ana Blanco,F.I.Javier Pastor.Xylanase production by a new lakalitolerant isolate of Bacilus[J].Biotecbnology Letter.1998,20(3):243-24.
[68]Anwat Sunna,Peter L.Bergquist.A gene encoding a novel extremely thermostable 1,4-β-xylanase isolated directly from an environmental DNA sample[J].ExtremopHiles.2003,7:63-70.
[69]M.Bataillon,A.P.Nunes Cardinali,N.Castillon,et al.Purification and characterization of a moderately thermostable xylanase from Bacillus so strain SPS-0[J].Enzymes and Microbial Technology.2000, 26:187-192.
[70]黄运红,龙中儿,李素珍等.木聚糖酶高产微生物的筛选和鉴定[J].江西师范大学学报(自然科学版),2002,26(3):245-248.
[71]李用芳,李学梅,张建新等.木聚糖酶生产菌的选育及发酵条件研究[J].生物技术,2004,14(1):25-27.
[72]许君,任艳丽,陈红歌等.碱性木聚糖酶产生菌的筛选与产酶条件[J].生物技术,2004,14(3):52-54.
[73]Lino Mayorga-Reyes,Teresa Ponce-Noyola.Isolation of a hyperxylanolytic Cellulomonas flavigena mu -tant growing on continuous culture on sugar cane bagasse[J].Biotechnology Letter, 1998,20(5): 443– 446.
[74]Rodrigo Pires do Nascimento,Susana Marques,Luis Alves,et al.A nobel strain of Streptomyces malaysiensis isolated from Brazilian soil produces high endo-β-1,4-xylanase titres[J].World Journal of Microbiology &Biotechnology,2003,19:879-881.
[75]Amita R,Shah Datta Madamwar.Xylanase production by a newly isolated Aspergillus foetidus strain and its characterization[J].Process Biochemistry, 2005,40:1763–1771.
[76]K.G. Sonia,B.S. Chadha,H.S. Saini.Sorghum straw for xylanase hyper-production by Therm- omyces lanuginosus (D2W3) under solid-state fermentation[J].Bioresource Technology,2005,96: 1561–1569.
[77]包怡红,刘伟丰,毛爱军等.耐碱性木聚糖酶高产菌株的筛选、产酶条件优化及其在麦草浆生物漂白中的应用[J].农业生物技术学报,2005,13(2):235-240.
[78]Zheng-Hong Xu,Yun-Ling Bai,Xia Xu,et al.Production of alkali-tolerant cellulose-free xylanase by Pseudomonas sp. WLUN024 with wheat bran as the main substrate[J].World Journal of Microbiology & Biotechnology,2005,21:575-581.
[79]郭勇主编.酶工程[M].科学出版社,1994.
[80]沈同主编.生物化学(第二版)[M].高等教育出版社,1991.
[81]袁志发主编.生物统计[M](第一版).高等教育出版社, 2000.
[82]J.萨姆布鲁克等著,金冬雁,黎孟枫等译.分子克隆实验指南(第三版)[M],科学出版社,1999.
[83]Michael Waino,Kjeld Ingvorsen.Production of β-xylanase and β-xylosidase by the extremely halopHilic archaeon Halothabdus utahensis[J].ExtremopHiles,2003,7:87-93.
[84]O.Gallardo,P.Diaz,F.I.J.Pastor.Characterization of a Paenibacillus cell-associated xylanase with high activity on aryl-xylosides:a new subclass of family 10 xylanases[J].Appl Microbiol Biotechnol, 2003, 61:226-233.
[85]Isil Seyis,Nilufer Aksoz.Effect of carbon and nitrogen sources on xylanase production by Trichoderma harzianum 1073 D3[J].International Biodeterioration & Biodegradation, 2005,55:115–119.
[86]Eleonora Cano Carmona, Mauricio Batista Fialho,érika Bicalho uchgnani,et al. Production,purifcation and characteri -zation of a minor form of xylanase from Aspergillus versicolor[J].Process Biochemistry, 2005,40:359–364.
[87]Bataillon M, Cardinali APN, Duchiron F.Production of xylanases from a newly isolated lkalopHilic thermopHilic Bacillus sp[J].Biotechnol Lett,1998,20:1067–1071.
[88]Beg QK, Bhushan B, Kapoor M, et al.Production and characterization of thermostable xylanase and pectinase from a Streptomyces sp. QG-11-3[J].J Ind Microbiol Biotechnol,2000a,24:396–402.
[89]Liu W, Lu Y, Ma G.Induction and glucose repression of endo-β-xylanase in the yeast Trichosporon cutaneu,m SL 409[J].Process Biochem.1999,34:67–72.
[90]Ishihara M, Tawata S, Toyama S.Purification and some properties of a thermostable xylanase from ther -mopHilic fungus strain HG-1[J].J Ferment Bioeng,1997,83:478–480.
[91]Xiong H. Xylanase production by Trichoderma reesei Rut C-30 grown on L-arabinose-rich plant hydro -lysates[J].Bioresource Technology,2005,96 (7):753 759.
[92]Licia Lama,Valeria Calandrelli,Agata Gambacorta,Barbara Nicolaus.Purification and characterization of thermostable xylanase and β-xylosidase by the thermopHilic bacterium Bacillus thermantarcticus[J]. Research in Microbiology,2004,155:283–289.
[93]S.Virupakshi,K.Gireesh Babu,Satish R Gaikwad,et al.Production of a xylanolytic enzyme by a thermo -alkalipHilic Bacillus sp. JB-99 in solid state fermentation[J].Process iochemistry,2005,40:431–435.
[94]Lopez C, Blanco A, Pastor FIJ .Xylanase production by a new alkali-tolerant isolate of Bacillus.[J] Biotechnol Letter.1998,20:243–246.
[95]Liu W, Zhu W, Lu Y, et al. Production, partial purification and characterization of xylanase from Trichosporon cutaneum SL409[J]. Process Biochem,1998,33:331–326.
[2]J.萨姆布鲁克,D.W.拉塞尔著,黄培堂等译.分子克隆实验指南(第三版)[M],科学出版社,2002
[3]郭勇等,酶工程[M].中国轻工业出版社,1997.
[4]周晨妍,邬敏辰.木聚糖酶的酶学特性与分子生物学[J].生物技术,2005,15(3):89-92.
[5]刘瑞田,曲音波,杨建云.木聚糖酶分子结构研究进展[J].纤维素科学与技术,1997,5(3):1-6.
[6]张红莲,姚斌,范云六.木聚糖酶的分子生物学及其应用[J].生物技术通报,2002,3:23-30.
[7]韩晓芳,郑连爽.产木聚糖酶嗜碱细菌的筛选及产酶条件研究[J].环境污染智力技术与设备,2002,3(11):25-27.
[8] 曾莹 , 熊志刚 , 李彦 . 啤酒糟产木聚糖酶的菌种筛选及其产酶条件研究 [J]. 粮油食品科技,2005,13(2):4-6.
[9]Barbara Decelle,Adrian Tsang,Reginald K.Storms.Cloning,functional expression and characterization of three PHanernchaete chrysosporium endo-1,4-β-xylanase [J].Current Genet,2004,46:166-175.
[10]Rifaat Rawashdeh,Ismail Saadoun,Amjad Mahasneh.Effect of cultural conditions on xylanase production by Streptomyces so.(strain ib 24D)and its potential to utilize tomato pomace[J].Arican Journal of Biotechnology,2005,4(3):251-255.
[11]H.Hayashi,M.Takehara,T.Hattori,et al.Nucleotide sequences of two contiguous and highlyhomologous xylanase genes xynA and xynB and characterization of XynA from Clostridium thermocellum.[J].Appl Microbiol Biotechnol,1999,(51):348-357.
[12]Satoshi Nakamura.Structure and function of a multidomain alkaline xylanase from alkalipHilic Bacillus sp.strain 41M-1[J].Catalysis Surveys from Asia, 2003,7(2):157-164.
[13]Suren Singh,Andreas M.Madlala,Bernard A.Prior.Thermomyces lanuginosus: properties of strains and their hemicellulase[J].FEMS Microbiology Reviews,2003,27:3-16.
[14]Badal C.Saha.Hemicellulose bioconversion[J].J Ind MicrobioI Biotechnol, 2003,30:279-291.
[15]Neeta Kulkarnnni,Abhay Shendye,Mala Rao.Molecular and biotechnological aspects of xylanases [J]. FEMS Microbiology Reviews,1999,23:411-456.
[16]Licia Lama,Valeria Calandrelli,Agata Gambacorta,et al.Purification and characterization of thermostab -le xylanase shan β-xylosidase by the thermopHilic bacterium Bacillus thermantarcticus[J].Research in Microbiology, 2004,155:283-289.
[17]Q.K.Beg,M.Kapoor,L.Mahajan,et al.Microbial xylanases and their industrial applications:a review [J]. Appl Microbiol Biotechnol,2001,56:326-338.
[18]Priya Chaudhary,D.N.Deobagkar.Characterization of Cloned Endoxylanase from Cellulomonas sp.NCI -M2353 Expressed in Escherichia coli[J].Current Microboilogy, 1997;34:273-279.
[19]P.P.Dwivedi,M.D.Gibbs,D.J.Saul,et al.Cloning,sequencing and over expressing in Escherichia coli of a xylanase gene,xynA from the thermopHilic bacterium Rt8B.4 genus Caldicelluslosiruptor[J].Appl Microbiol Biotechnol, 1996,45:86-93.
[20]Oscar Gallardo,Pilar Diaz,F.I.Javier Pastor.Cloning and Characterization of Xylanase A from the strain Bacillus sp.BP-7:Comparision with Alkaline pI-Low Molecular Weight Xylanases of Family 11[J]. Cur -rent Microbiology,2004,48:276-279.
[21]O.Gallardo,P.Diaz,F.I.J.Pastor.Charaterization of a Paenibacillus cell-associa ted xylanase with highactivity on aryl-xylosides:a mew subclass of family 10 xylanases[J].Appl Microbiol Biotechnol , 2003,61: 226-233.
[22]Mursheda K.Ali,Frederick B.RudolpH,George N.Bennett.Characterization of thermostable Xyn10A enzyme form mesopHilic Clostridiun acetobutylicum ATCC824[J].Ind Microbiol Biotechnol,2005,32: 12-18.
[23]杨翔华,洪新.产木聚糖酶菌株的筛选及其产酶特性[J].辽宁石油化工大学学报,2004,24(4):32-35.
[24]Mursheda K.Ali,Frederick B.RudolpH,George N.Bennett.Thermostable xulanase 10B from Clostridiun acetobutylicum ATCC824[J].J Ind Microbiol Biotechnol,2004,31: 229-234.
[25]Hyeon Jin Sun,Shigeki Yoshida,Yasuyuki Kawabata,et al.Separatuin of two functiuonal domains of the family F/10 β-xylanase from Steptomyces olicaceociridis E-86 limited oeoteolysis with papain and some of their properties[J].Biotechnology Letter,2002,24:595-601.
[26]Hyun S Jun,Jong K.Ha,Laercio M.Malburg et al.Charateristics of a cluster of xylanase genes in Fibrobacter succinogenes S85[J].Canadian Journal of Microbiology,2003,49 (3):171-180.
[27]M.Bztaillon,AP.Nunes Carsinali,F.Duchiron.Production of xylanases from a newly isolated alkalopHiic thermopHilic Bacillus sp[J].Biotechbnology Letter,1998,20(11): 1067-1071.
[28]Laurie JI,Clarke JH,Ciruela A,et al.The Nodb domain of a multidomain xylanases from Cellulomonas fimi deacetylates acetlxylan.FEMS Letter,1997,148:261-264.
[29]Daniel D.Morris,Moreland N.Gibbs,Michelle Ford,et al. Family 10 and 11 xylanase genes from Caldicellulosiruptor sp.strain RT69B.1 [J].ExtermopHiles.1999,3:103-111.
[30]A .Shlacher,K Holzmann,M.Hayn,et al.Cloning and characterization of the gene for the thermostable xylanase XynA from Thermomyced lanuginosus[J].Journal of Biotechnology.1994,49:211-218.
[31]Hania Kebir,Claude Dupont,Rolf Morosoli.Increased xylanase production in Streptomyces lividans after replacement of the signal peptide:dependence on box and inverted repeat sequence[J].Biochimica et BiopHysica Acta,2000,1491:177-184.
[32]Asbah F.Qureshy,L A.Khan,S.Khanna.Expression of Bacillus circulans Teri-42 xylanase gene in Bacillus subtilis[J].Enzyme and Microbial Technology.2000,27:227-233.
[33]刘明启,孙建义,许英蕾.木聚糖酶分子进化的研究进展[J].中国生物工程杂志,2003,23(10):62-66.
[34]丁少军,J.A.BUSWELL.利用真菌纤维素结合域(CBD)保守性序列进行草菇木聚糖酶cDNA 的克隆[J].中国生物化学与分子生物学报,2004,20(3):408-412.
[35]T.Kmiura,T.Mizutani,T.Tanaka,et,al.Molecular breeding of transgenic rice expressing a xylanase doma -in of the xynA gene from Clostridium thermmocellum[J].Appl Microbiol Biotechnol, 2003,(62): 374- 379.
[36]G.P.Xue.J.S.Johnson,D.J.Smyth et al.Temperature-regulated expression of the tac/lacI system for overproduction of a fungal xylanase in Escherichia coli[J].Appl Microbiol Biotechnol.1996, 45:120-126.
[37]H.Kim,H.Jong,M.Y.Pack..Molecular characterization of xynX,a gene encoding a multidomain xylanase with a thermostabilizing domain form Clostridium thermocellum[J] .Appl Microbiol Biotechnol (2000) 54:521-527.
[38]Sirin S.I.Adham,Pilar Honrubia,Matgarita Diaz,et al.Expression of the genes coding for the xylanase Xys1 and the cellulose Cel1 from the straw-decompising Streptomyces halstedii JM8 cloned into the amino acid peoducet Brevibacterium lactofermentum ATCC13869[J].Arch Microbiol.201,177:91-97.
[39]Kaveh Emami,Ethan Hack.Conservation of XYN11A and XYN11B Xylanase Genes in Bipolaris sorghicola,Cochliobolus sativus, Cochliobolus heterostropHus,and Cochlo\iobilus specifer[J].Current Microbiology,2005:45:303-306.
[40]M.Uma Maheswari,T.S.Chandra.Production and potential application of a xylanas from a new strain of Streptomyces cuspidosporus[J].World Journal of Microbiology & Biotechnology,2000,16:257-263.
[41]Chartchai Khanongnuch,Saidamim Lumyong,Toshihiko Ooi,et al.A non-cellulase producing strain of Bacillus subtilis and its potential use in pulp biobleaching.Biotechnology Letter,1999,21:61-63.
[42]F.Fernadez,R.Sharma,M.Hinton,et al.Diet influences the colonidation of Campylobacter jejuni and distribution of mucin carbohydrates in the chick intestinal tract [J].Cell. Mol.Life. Science, 2000, 57: 1793-1801.
[43]J.H.Clarke,J.E.Rixon,A.Ciruela,et al.Family-10 and Family-11 xylanase differ in their capacity to enha -nce the bleachability of hardwoof and softwood paper pulps[J].Appl Microbiol Biotechnol , 1997, 48: 177-183.
[44]Chandralata Raghukumar,Usha Muraleedhara n,V.R.Gaud,et al.Xylanase of marine fungi of potential use for biobleaching of paper pulp[J].J Ind Microbiol Biotechnol,2004,31:433-441.
[45]何为,詹怀宇.木聚糖酶酶活影响因素的研究[J].中国造纸学报,2009,19(1):163-166.
[46]苏纯阳,程学慧,刘涛等.新一代的饲用酶制剂-细菌性木聚糖酶[J].饲料工业,2004,12:41-43.
[47]王金全,蔡辉益,张姝等.木聚糖酶对0-3周龄肉仔鸡肠道食糜黏度和生产性能的影响[J].中国家禽学报,2004,8(1):122-125.
[48]刘明启,孙建义,郭爽.木聚糖酶嗜酸性、热稳定性研究进展[J].中国饲料,2004,13:21-22.
[49]吴建忠,李绿雄.木聚糖酶在畜禽营养中的研究及应用[J].农业与技术,2005,25(2):73-77.
[50]方俊.木聚糖酶在猪饲料中的研究与应用[J].中国畜牧兽医,2003,30(3):20-22.
[51]王修启,林东康.皖麦38添加木聚糖酶对鸡表观代谢能值和氨基酸表观消化率的影响[J].华北农学报,2003,18(1):116-118.
[52]王海英,呙于明,袁建敏.小麦日粮中添加木聚糖酶对肉仔鸡生产性能和养分消化率的影响[J].粮食与饲料工业,2003,12:53-55.
[53]夏枚生.高麸日粮中添加复合酶制剂对仔猪血液中几种激素水平的影响[J].浙江农业科学,2000,6:300-303.
[54]王振来,钟艳玲,路广计等.木聚糖酶、β-葡聚糖酶和纤维素酶促进仔猪生长机理探讨[J].中国饲料,2004,9:20-23.
[55]Yunben B.Wu,Velmurugu Ravindran. Influence of whole wheat inclusion and xylanase supplementati- on on the performance, digestive tract measurements and carcass characteristics of broiler chickens[J]. Animal Feed Science and Technology,2004,116:129–139.
[56]Ravindran, V., Cabahug, S., Ravindran, et al. Influence of microbial pHytase on apparent ileal amino acid digestibility in feedstuffs for broilers[J]. Poult. Sci. 1999,78:699–706.
[57]F.Fernadez,R.Sharma,M.Hinton,et al.Diet influences the coloniation of Campylobacter jejuni and dis -tribution of mucin carbohydrates in the chick intestinal tract[J] Cell Mol.Life.Science, 2000,57: 1793- 1801.
[58]Ashita Dhillon,J.KGupta,B.M.Jauhari,et al.A celluase–poor thermstable,alkalitolerant xylanase produ -ced by Bacillus circulans AB16 grown on rice straw ans its application in biobleaching of eucalyptus pulp[J].Bioresource Technology.2000,(73):273-277.
[59]Chandralata Raghukumar,Usha Muraleedharan V.R,Gaud R.Mishra. Xylanase of marine fungi of potential use for biobleaching of paper pulp[J].J Ind Microbiol Biotechnol,2004,31: 433-441.
[60]Sunita Chauhan,B.Choudhury,S.N. Singh,et al.Application of xylanase enzyme of Bacillus coagulans as a prebleaching agent on non-woody pulps[J].Process Biochemistry, 2006,41 226–231.
[60]Chartchai Khanongnuch,Saidamim Lumyong,Toshihiko Ooi,et al.A non-cellulase producing strain of Bacillus subtilis and its potential use in pulp biobleaching.Biotechnology Letter,1999,21:61-63.
[61]M. Blanca Roncero,Antonio L. Torres,Jos_e F. Colom,et al.The effect of xylanase on lignocellulosic components during the bleaching of wood pulps[J].Bioresource Technology,2005, 96:21–30.
[62]J.H.Clarke,J.E.Rixon,A.Ciruela,H.J.Gilbert,G.P.Hazlewood.Family-10 and Family-11 xylanase differ in their capacity to enhance the bleachability of hardwoof and softwood paper pulps[J].Appl Microbiol Biotechnol ,1997,48:177-183.
[63]Xiaoping Yuan,Jing Wang,Huiyuan Yao.Production of feruloyl oligosaccharides from wheat bran insoluble dietary fibre by xylanases from Bacillus subtilis[J].Food Chemistry,2006,95:484–492.
[64]I.Trogh,C.M. Courtin,A.A.M.Andersson,P.A man,et al. Delcour.The combined use of hull-less barley flour and xylanase as a strategy for wheat/hull-less barley flour breads with increased arabinoxylan and (1/3,1/4)-b-D-glucan levels[J].Journal of Cereal Science, 2004,40:257–267.
[65]Tony Collins,Anne Hoyoux,Agne`s Dutron,et al.Use of glycoside hydrolase family 8 xylanases in baking[J].Journal of Cereal Science,2006,43:79–84.
[66]沈萍,范容秀,李广武主编.微生物实验[M].高等教育出版社(第三版),1999,6.
[67]Cristina Lopez,Ana Blanco,F.I.Javier Pastor.Xylanase production by a new lakalitolerant isolate of Bacilus[J].Biotecbnology Letter.1998,20(3):243-24.
[68]Anwat Sunna,Peter L.Bergquist.A gene encoding a novel extremely thermostable 1,4-β-xylanase isolated directly from an environmental DNA sample[J].ExtremopHiles.2003,7:63-70.
[69]M.Bataillon,A.P.Nunes Cardinali,N.Castillon,et al.Purification and characterization of a moderately thermostable xylanase from Bacillus so strain SPS-0[J].Enzymes and Microbial Technology.2000, 26:187-192.
[70]黄运红,龙中儿,李素珍等.木聚糖酶高产微生物的筛选和鉴定[J].江西师范大学学报(自然科学版),2002,26(3):245-248.
[71]李用芳,李学梅,张建新等.木聚糖酶生产菌的选育及发酵条件研究[J].生物技术,2004,14(1):25-27.
[72]许君,任艳丽,陈红歌等.碱性木聚糖酶产生菌的筛选与产酶条件[J].生物技术,2004,14(3):52-54.
[73]Lino Mayorga-Reyes,Teresa Ponce-Noyola.Isolation of a hyperxylanolytic Cellulomonas flavigena mu -tant growing on continuous culture on sugar cane bagasse[J].Biotechnology Letter, 1998,20(5): 443– 446.
[74]Rodrigo Pires do Nascimento,Susana Marques,Luis Alves,et al.A nobel strain of Streptomyces malaysiensis isolated from Brazilian soil produces high endo-β-1,4-xylanase titres[J].World Journal of Microbiology &Biotechnology,2003,19:879-881.
[75]Amita R,Shah Datta Madamwar.Xylanase production by a newly isolated Aspergillus foetidus strain and its characterization[J].Process Biochemistry, 2005,40:1763–1771.
[76]K.G. Sonia,B.S. Chadha,H.S. Saini.Sorghum straw for xylanase hyper-production by Therm- omyces lanuginosus (D2W3) under solid-state fermentation[J].Bioresource Technology,2005,96: 1561–1569.
[77]包怡红,刘伟丰,毛爱军等.耐碱性木聚糖酶高产菌株的筛选、产酶条件优化及其在麦草浆生物漂白中的应用[J].农业生物技术学报,2005,13(2):235-240.
[78]Zheng-Hong Xu,Yun-Ling Bai,Xia Xu,et al.Production of alkali-tolerant cellulose-free xylanase by Pseudomonas sp. WLUN024 with wheat bran as the main substrate[J].World Journal of Microbiology & Biotechnology,2005,21:575-581.
[79]郭勇主编.酶工程[M].科学出版社,1994.
[80]沈同主编.生物化学(第二版)[M].高等教育出版社,1991.
[81]袁志发主编.生物统计[M](第一版).高等教育出版社, 2000.
[82]J.萨姆布鲁克等著,金冬雁,黎孟枫等译.分子克隆实验指南(第三版)[M],科学出版社,1999.
[83]Michael Waino,Kjeld Ingvorsen.Production of β-xylanase and β-xylosidase by the extremely halopHilic archaeon Halothabdus utahensis[J].ExtremopHiles,2003,7:87-93.
[84]O.Gallardo,P.Diaz,F.I.J.Pastor.Characterization of a Paenibacillus cell-associated xylanase with high activity on aryl-xylosides:a new subclass of family 10 xylanases[J].Appl Microbiol Biotechnol, 2003, 61:226-233.
[85]Isil Seyis,Nilufer Aksoz.Effect of carbon and nitrogen sources on xylanase production by Trichoderma harzianum 1073 D3[J].International Biodeterioration & Biodegradation, 2005,55:115–119.
[86]Eleonora Cano Carmona, Mauricio Batista Fialho,érika Bicalho uchgnani,et al. Production,purifcation and characteri -zation of a minor form of xylanase from Aspergillus versicolor[J].Process Biochemistry, 2005,40:359–364.
[87]Bataillon M, Cardinali APN, Duchiron F.Production of xylanases from a newly isolated lkalopHilic thermopHilic Bacillus sp[J].Biotechnol Lett,1998,20:1067–1071.
[88]Beg QK, Bhushan B, Kapoor M, et al.Production and characterization of thermostable xylanase and pectinase from a Streptomyces sp. QG-11-3[J].J Ind Microbiol Biotechnol,2000a,24:396–402.
[89]Liu W, Lu Y, Ma G.Induction and glucose repression of endo-β-xylanase in the yeast Trichosporon cutaneu,m SL 409[J].Process Biochem.1999,34:67–72.
[90]Ishihara M, Tawata S, Toyama S.Purification and some properties of a thermostable xylanase from ther -mopHilic fungus strain HG-1[J].J Ferment Bioeng,1997,83:478–480.
[91]Xiong H. Xylanase production by Trichoderma reesei Rut C-30 grown on L-arabinose-rich plant hydro -lysates[J].Bioresource Technology,2005,96 (7):753 759.
[92]Licia Lama,Valeria Calandrelli,Agata Gambacorta,Barbara Nicolaus.Purification and characterization of thermostable xylanase and β-xylosidase by the thermopHilic bacterium Bacillus thermantarcticus[J]. Research in Microbiology,2004,155:283–289.
[93]S.Virupakshi,K.Gireesh Babu,Satish R Gaikwad,et al.Production of a xylanolytic enzyme by a thermo -alkalipHilic Bacillus sp. JB-99 in solid state fermentation[J].Process iochemistry,2005,40:431–435.
[94]Lopez C, Blanco A, Pastor FIJ .Xylanase production by a new alkali-tolerant isolate of Bacillus.[J] Biotechnol Letter.1998,20:243–246.
[95]Liu W, Zhu W, Lu Y, et al. Production, partial purification and characterization of xylanase from Trichosporon cutaneum SL409[J]. Process Biochem,1998,33:331–326.