克氏原螯虾N-乙酰-β-D-氨基葡萄糖苷酶的性质及活力调控
|
摘要
N-乙酰-β-D-氨基葡萄糖苷酶在昆虫和甲壳动物角质层的转化和转移及营养消化的生理过程中发挥了重要作用。本研究采用硫酸铵分级沉淀从克氏原螯虾内脏分离出粗酶制剂后再经前后2次DEAE-32离子交换柱层析和SephadexG-100分子筛柱层析获得比活力为6540.24 U·mg-1的聚丙烯酰胺凝胶电泳纯N-乙酰-β-D-氨基葡萄糖苷酶纯酶制剂。测得该酶分子的总分子量为118.66 kDa,等电点为5.15 ,每个酶分子中含有约400余个氨基酸残基;以对-硝基苯-N-乙酰-β-D-氨基葡萄糖(pNP-β-D-GlcNAc)为底物,研究酶催化底物水解的反应动力学,结果表明,酶的最适温度为40℃,最适pH为5.5。该酶在50℃以下处理30 min,酶活力稳定,高于55℃,酶稳定性较差, 75℃酶完全失活;在pH 3.6~5.9范围内较稳定,而在pH>6.2和pH<3.6下失活加快;酶催化底物反应的活化能为114.89 kJ·mol-1。以对-硝基苯-N-乙酰-β-D-氨基葡萄糖(pNP-β-D-GlcNAc)和p-硝基苯基-N-乙酰-β-D-氨基半乳糖苷(pNP-β-D-GaNAc)为底物,研究酶促反应动力学符合米氏双曲线方程,测得以pNP-β-D-GlcNAc为底物时最大反应速度Vm为10.87μmol·L-1min-1,米氏常数Km为0.46 mmol·L-1;以pNP-β-D-GaNAc为底物时最大反应速度Vm为39.40μmol·L-1min-1,米氏常数Km为1.04 mmol·L-1。
金属离子对克氏原螯虾NAGase的效应试验表明:在特定浓度范围内, Li+ ,K+对酶活力几乎没有影响; Na+、Ba2+、Ca2+、Co2+则对酶有不同程度的激活作用;Mn2+、Mg2+、Pb2+、Ag+对酶有先扬后抑的作用;Zn2+、Hg2+、Cu2+、Al3+等重金属离子对酶都有不同程度的抑制作用。在一定浓度范围内,金属离子对克氏原螯虾NAGase的抑制作用强度从大到小依次为:Hg+ > Cu2+ > Al3+ > Zn2+ > Ag+ > Pb2+ > Mn2+ > Mg2+。Mn2+、Ag+、Zn2+、Al3+、Hg+对克氏原螯虾NAGase的抑制均表现为可逆反竞争性机制,其抑制常数KI分别为0.41 mmol·L-1、1.08 mmol·L-1、1.22 mmol·L-1、0.87 mmol·L-1、0.069 mmol·L-1。Mg2+和Pb2+对克氏原螯虾NAGase的抑制表现分别为竞争性和非竞争性,其抑制常数KI分别为1.59 mmol·L-1和0.41 mmol·L-1。
氨基酸对克氏原螯虾NAGase的效应试验表明:在特定浓度范围内,非极性氨基酸L-Ala、L-Phe、L-Met、L-Pro,极性氨基酸L-Gly、L-Thr、L-Ser、L-Gln对克氏原螯虾NAGase活力没有显著影响,碱性氨基酸L-Arg、L-His、L-Lys对克氏原螯虾NAGase有不同程度的抑制作用。L-Arg、L-His、L-Lys对克氏原螯虾NAGase的IC50为45mmol·L-1、130mmol·L-1、110mmol·L-1。碱性氨基酸L-Arg、L-His、L-Lys对克氏原螯虾NAGase的抑制均表现为可逆非竞争性,其KI分别为10.76 mmol·L-1、226.12 mmol·L-1、157.70 mmol·L-1。
研究不同生长阶段克氏原螯虾各器官NAGase活力变化。结果表明:克氏原螯虾肝胰腺NAGsase活力随着年龄的增大逐期递减。酶活力I期(2~3月龄)与II期(5~6月龄)比较降低趋势差异显著,III期(9~10月龄)和II期比较有降低趋势,但差异不明显;克氏原螯虾性腺和壳膜NAGase活力变化规律一致,都是逐期递减,但I期和II期递减趋势不明显,到了III期后有了明显的下降。此外,比较同一生长期克氏原螯虾各器官NAGsase活力大小顺序为:肝胰腺>性腺>壳膜。
比较研究不同生长阶段克氏原螯虾肝胰腺、性腺及甲壳膜三个器官NAGase的理化性质,发现克氏原螯虾肝胰腺NAGase在I、II、III期的最适温度都是35℃;性腺NAGase在I、II期最适温度为35℃,III期为40℃;甲壳膜NAGase在I期最适温度为40℃,II、III期均为35℃。克氏原螯虾肝胰腺NAGase在I、III期的最适pH都是5.5,II期为5.2;性腺NAGase在I期最适pH为5.5,II、III期为5.2;甲壳膜NAGase在I、II期的最适pH均为4.5,在III期为5.5。
β-N-Acetyl-D-glucosaminidase play an important role in the physiological process the conversion and transformation of the cuticle of insects ,crustaceans and nutrition digestion.
Theβ-N-Acetyl-D-glucosaminidase was purified from Procambarus clarkii by ammonium sulfate fractionation, twice tchromatography on DEAE-cellulose (DEAE-32) and Sephadex G-100.The purified enzyme showed a single band on polyacrylamide gel electrophoresis and isoelectric focusing electrophoresis,and the specific activity was 6540.24 U·mg -1.The molecular weight of the whole enzyme was 118.66kD, consisted of 364 amino acid residues.Its isoelectric point is 5.15. The optimum pH and optimum temperature of the enzyme for the hydrolysis of p-nitrophenyl-N acetyl-β-D-glucosaminide (pNP-β-D-GlcNAc)were found at pH 5.5 and at 40°C, respectively. The study of its stability showed that the enzyme was stable in the pH range from 3.6 to 5.9 and at temperatures below 50°C. The activation energy was 114.89kJ·mol-1.The kinetic behavior of the enzyme in the hydrolysis of pNP-β-D-GlcNAc followed Michaelis–Menten kinetics with Km of 0.46 mmol·L-1 and Vm of 10.87μmol·L-1·min -1 ,and the kinetic behavior of the enzyme in the hydrolysis of pNP-β-D-GaNAc followed Michaelis–Menten kinetics with Km of 1.04 mmol·L-1 and Vm of 39.40μmol·L-1·min -1 .
The effects of metal ions on the enzyme were studied. The results showed that Li2+、K+ had not any effect on enzyme activity in the tested range of concentration. Na+、Ba2+、Ca2+、Co2+ could increase the activity of enzyme at different concentration. Mn2+、Mg2+、Pb2+、Ag+ increase enzyme activity at first then inhibit it. Zn2+、Hg2+、Cu2+、Al3+ were significantly reduce enzyme activity to different degrees. The intensity of inhibition effects of metal ions on the enzyme from big to small in turn: Hg+ > Cu2+ > Al3+ > Zn2+ > Ag+ > Pb2+ > Mn2+ > Mg2+. Mn2+、Ag+、Zn2+、Al3+、Hg+ were reversible anticompetitive inhibitor . The KI were 0.41 mmol·L-1、1.08 mmol·L-1、1.22 mmol·L-1、0.87 mmol·L-1、0.07 mmol·L-1 respectively. Mg2+ was revealed to be a reversible non-competitive inhibitor with a KI of 1.59 mmol?L-1.Pb2+ was revealed to be a reversible competitive inhibitor with a KI of 26.17 mmol?L-1.
The effects of amino acids on the enzyme were studied. The results showed that L-Ala、L-Phe、L-Met、L-Pro、L-Gly、L-Thr、L-Ser、L-Gln had not any effect on enzyme activity in the tested range of concentration. L-Arg、L-His、L-Lys reduced the activity of enzyme at different concentration.The IC50 of L-Arg、L-His、L-Lys on the enzyme in turn: 45mmol·L-1、130mmol·L-1、110mmol·L-1. L-Arg、L-His、L-Lys were revealed to be a reversible competitive inhibitor with KI of 10.76mmol?L-1、226.12 mmol?L-1、.157.70 mmol?L-1.
We studied the variations of activity ofβ-N-Acetyl-D-glucosaminidase from different organs of Procambarus clarkii in different growth stages. The result s showed that the enzyme activities of the hepatopancreas of Procambarus clarkii in different growth stages are variational.The activity of enzyme from the hepatopancreas of Procambarus clarkii was decreased one by one stage,there was a significant decreasing in stage I(2~3moon’s age) and stage II(5~6 moon’s age)、stage III(9~10 moon’s age).But had no significantly decreased in stageII and stage III . The activity of enzyme of the gonad were consistent with variation of shell membrane: decreasing one by one stage ,but there were no significantly decreased between in stageII and stageIII,significantly decreased in stageIII . In addition, the results showed that the enzyme activities of the variations organs of Procambarus clarkii from big to small in turn:The hepatopancreas> The gonad >The shell membrane.
Studied some basic properties ofβ-N-Acetyl-D-glucosaminidase from different organs of Procambarus clarkii in different growth stages. The result s showed that the optimum temperature of the enzyme from the hepatopancreas of Procambarus clarkii is 35℃in all stage; the optimum temperature of the activity of enzyme of the gonad were 35℃in stageI、stageII.but 40℃in stageIII.The optimum temperature of the activity of enzyme of of shell membrane were 35℃in stageII、stageIII.but 40℃in stageI. The optimum pH of the enzyme from the hepatopancreas were 5.5 in stageI、stageIII. but 5.2 in stageII;The optimum pH of the activity of enzyme of the gonad were 5.2 in stageII、stageIII,but 5.5 in stageI; The optimum pH of the activity of enzyme of of shell membrane were 4.5 in stageI、stageII,but5.5 in stageIII.
金属离子对克氏原螯虾NAGase的效应试验表明:在特定浓度范围内, Li+ ,K+对酶活力几乎没有影响; Na+、Ba2+、Ca2+、Co2+则对酶有不同程度的激活作用;Mn2+、Mg2+、Pb2+、Ag+对酶有先扬后抑的作用;Zn2+、Hg2+、Cu2+、Al3+等重金属离子对酶都有不同程度的抑制作用。在一定浓度范围内,金属离子对克氏原螯虾NAGase的抑制作用强度从大到小依次为:Hg+ > Cu2+ > Al3+ > Zn2+ > Ag+ > Pb2+ > Mn2+ > Mg2+。Mn2+、Ag+、Zn2+、Al3+、Hg+对克氏原螯虾NAGase的抑制均表现为可逆反竞争性机制,其抑制常数KI分别为0.41 mmol·L-1、1.08 mmol·L-1、1.22 mmol·L-1、0.87 mmol·L-1、0.069 mmol·L-1。Mg2+和Pb2+对克氏原螯虾NAGase的抑制表现分别为竞争性和非竞争性,其抑制常数KI分别为1.59 mmol·L-1和0.41 mmol·L-1。
氨基酸对克氏原螯虾NAGase的效应试验表明:在特定浓度范围内,非极性氨基酸L-Ala、L-Phe、L-Met、L-Pro,极性氨基酸L-Gly、L-Thr、L-Ser、L-Gln对克氏原螯虾NAGase活力没有显著影响,碱性氨基酸L-Arg、L-His、L-Lys对克氏原螯虾NAGase有不同程度的抑制作用。L-Arg、L-His、L-Lys对克氏原螯虾NAGase的IC50为45mmol·L-1、130mmol·L-1、110mmol·L-1。碱性氨基酸L-Arg、L-His、L-Lys对克氏原螯虾NAGase的抑制均表现为可逆非竞争性,其KI分别为10.76 mmol·L-1、226.12 mmol·L-1、157.70 mmol·L-1。
研究不同生长阶段克氏原螯虾各器官NAGase活力变化。结果表明:克氏原螯虾肝胰腺NAGsase活力随着年龄的增大逐期递减。酶活力I期(2~3月龄)与II期(5~6月龄)比较降低趋势差异显著,III期(9~10月龄)和II期比较有降低趋势,但差异不明显;克氏原螯虾性腺和壳膜NAGase活力变化规律一致,都是逐期递减,但I期和II期递减趋势不明显,到了III期后有了明显的下降。此外,比较同一生长期克氏原螯虾各器官NAGsase活力大小顺序为:肝胰腺>性腺>壳膜。
比较研究不同生长阶段克氏原螯虾肝胰腺、性腺及甲壳膜三个器官NAGase的理化性质,发现克氏原螯虾肝胰腺NAGase在I、II、III期的最适温度都是35℃;性腺NAGase在I、II期最适温度为35℃,III期为40℃;甲壳膜NAGase在I期最适温度为40℃,II、III期均为35℃。克氏原螯虾肝胰腺NAGase在I、III期的最适pH都是5.5,II期为5.2;性腺NAGase在I期最适pH为5.5,II、III期为5.2;甲壳膜NAGase在I、II期的最适pH均为4.5,在III期为5.5。
β-N-Acetyl-D-glucosaminidase play an important role in the physiological process the conversion and transformation of the cuticle of insects ,crustaceans and nutrition digestion.
Theβ-N-Acetyl-D-glucosaminidase was purified from Procambarus clarkii by ammonium sulfate fractionation, twice tchromatography on DEAE-cellulose (DEAE-32) and Sephadex G-100.The purified enzyme showed a single band on polyacrylamide gel electrophoresis and isoelectric focusing electrophoresis,and the specific activity was 6540.24 U·mg -1.The molecular weight of the whole enzyme was 118.66kD, consisted of 364 amino acid residues.Its isoelectric point is 5.15. The optimum pH and optimum temperature of the enzyme for the hydrolysis of p-nitrophenyl-N acetyl-β-D-glucosaminide (pNP-β-D-GlcNAc)were found at pH 5.5 and at 40°C, respectively. The study of its stability showed that the enzyme was stable in the pH range from 3.6 to 5.9 and at temperatures below 50°C. The activation energy was 114.89kJ·mol-1.The kinetic behavior of the enzyme in the hydrolysis of pNP-β-D-GlcNAc followed Michaelis–Menten kinetics with Km of 0.46 mmol·L-1 and Vm of 10.87μmol·L-1·min -1 ,and the kinetic behavior of the enzyme in the hydrolysis of pNP-β-D-GaNAc followed Michaelis–Menten kinetics with Km of 1.04 mmol·L-1 and Vm of 39.40μmol·L-1·min -1 .
The effects of metal ions on the enzyme were studied. The results showed that Li2+、K+ had not any effect on enzyme activity in the tested range of concentration. Na+、Ba2+、Ca2+、Co2+ could increase the activity of enzyme at different concentration. Mn2+、Mg2+、Pb2+、Ag+ increase enzyme activity at first then inhibit it. Zn2+、Hg2+、Cu2+、Al3+ were significantly reduce enzyme activity to different degrees. The intensity of inhibition effects of metal ions on the enzyme from big to small in turn: Hg+ > Cu2+ > Al3+ > Zn2+ > Ag+ > Pb2+ > Mn2+ > Mg2+. Mn2+、Ag+、Zn2+、Al3+、Hg+ were reversible anticompetitive inhibitor . The KI were 0.41 mmol·L-1、1.08 mmol·L-1、1.22 mmol·L-1、0.87 mmol·L-1、0.07 mmol·L-1 respectively. Mg2+ was revealed to be a reversible non-competitive inhibitor with a KI of 1.59 mmol?L-1.Pb2+ was revealed to be a reversible competitive inhibitor with a KI of 26.17 mmol?L-1.
The effects of amino acids on the enzyme were studied. The results showed that L-Ala、L-Phe、L-Met、L-Pro、L-Gly、L-Thr、L-Ser、L-Gln had not any effect on enzyme activity in the tested range of concentration. L-Arg、L-His、L-Lys reduced the activity of enzyme at different concentration.The IC50 of L-Arg、L-His、L-Lys on the enzyme in turn: 45mmol·L-1、130mmol·L-1、110mmol·L-1. L-Arg、L-His、L-Lys were revealed to be a reversible competitive inhibitor with KI of 10.76mmol?L-1、226.12 mmol?L-1、.157.70 mmol?L-1.
We studied the variations of activity ofβ-N-Acetyl-D-glucosaminidase from different organs of Procambarus clarkii in different growth stages. The result s showed that the enzyme activities of the hepatopancreas of Procambarus clarkii in different growth stages are variational.The activity of enzyme from the hepatopancreas of Procambarus clarkii was decreased one by one stage,there was a significant decreasing in stage I(2~3moon’s age) and stage II(5~6 moon’s age)、stage III(9~10 moon’s age).But had no significantly decreased in stageII and stage III . The activity of enzyme of the gonad were consistent with variation of shell membrane: decreasing one by one stage ,but there were no significantly decreased between in stageII and stageIII,significantly decreased in stageIII . In addition, the results showed that the enzyme activities of the variations organs of Procambarus clarkii from big to small in turn:The hepatopancreas> The gonad >The shell membrane.
Studied some basic properties ofβ-N-Acetyl-D-glucosaminidase from different organs of Procambarus clarkii in different growth stages. The result s showed that the optimum temperature of the enzyme from the hepatopancreas of Procambarus clarkii is 35℃in all stage; the optimum temperature of the activity of enzyme of the gonad were 35℃in stageI、stageII.but 40℃in stageIII.The optimum temperature of the activity of enzyme of of shell membrane were 35℃in stageII、stageIII.but 40℃in stageI. The optimum pH of the enzyme from the hepatopancreas were 5.5 in stageI、stageIII. but 5.2 in stageII;The optimum pH of the activity of enzyme of the gonad were 5.2 in stageII、stageIII,but 5.5 in stageI; The optimum pH of the activity of enzyme of of shell membrane were 4.5 in stageI、stageII,but5.5 in stageIII.
引文
[1] AUSTIN P R. Chitin,Chitoson and Related Enzymes [M]. New York: Academic Press,1984.
[2]KEYHANI N O,WANG L X,LEE Y C.The chitin catabolic cascade in the marine bacterium Vibrio furnissii-Characterization of an N,N’-diacetyl-chitobiose transport system[J].Journal of Biological Chemistry,1996,271(52):33409~33413.
[3]BOLLER J,GEHRI A,MAUCH F.Chitinase in bean leaves:induction by ethylene,purification,properties,and possible function[J].Planta,1983,157:22~31.
[4]WERRIES E,NEUE I,BUDDECKE E.Evidence for different glycohydrolase and glycosyltransferase activities of beta-N-acetylglucosaminidases A and B[J].Hoppe Seylers Z Physiol Chem,1975,356(6):953~960.
[5]KONDO K,MATSUMOTO M,KOJO A,et al.Purification and characterization of chitinase from pupae Pieris rapae crucivora(Boiduval)[J].Journal of Chemical and Engineering of Japan,2002,35:241~246.
[6]USUI T,HAYASHI Y,NANJO F,et al.Transglycosylation reaction of a chitinase purified from Nocardia orientalis[J].Biochim Biophys Acta,1987,923(8):302~309.
[7]TAKEGAWA K,FUJITA K,FAN J Q,et al..Enzymatic synthesis of a neoglycoconjugate by transglycosylation with Arthrobacter endo-β-N-acetylglucosaminidase:A substrate for colorimetric detection of endo-β-N-acetylglucosaminidase activity.Analytical Biochemistry[J].1998,257(2):218~223.
[8] LIN JIAN CHENG, CHEN Q X, SHI Y,et al. The chemical modification of the essential groups ofβ-N-Acetyl-D-glucosaminidase from Turbo cornutus Solander[J]. IUBMB Life,2003,55 (9): 547~552.
[9] ZOU E,FINGERMAN M. Chitobiase activity in the epidermis and hepatopancreas of the fiddler crab Uca pugilator during the molting cycle[J]. Marine Biology,1999,133: 97~101.
[10]高磊,蔡国平.小鼠几丁质酶的重组表达和抗体制备[J].细胞与分子免疫学杂志,2008 9(25):9
[11]YOSHIDA K,UCHIJIMA Y, KOBAYASHI N,et al. Separation and properties of N-acetyl-beta-D-glucosaminidase from human seminal plasma . Article in Japanese[J] Nippon Hinyokika Gakkai Zasshi.1989 Nov;80(11):1625~31.
[12]YOSHIDA K,TAKAHASHI T,NAKAME Y ,et al.N-acetyl-beta-D-glucosaminidase (NAG) activity in human semen: its relation to gamma-glutamyl transpeptidase (gamma-GTP) activity in seminal plasma and reproductive tissues,and relation between seminal mucoprotein concentration and seminal NAG,and gamma-GTP activities[J]. Hinyokika Kiyo.1987 Jul;33(7):1054~9.
[13] EUR J CLIN CHEM CLIN BIOCHEM. Measurement of N-acetyl-beta-glucosaminidase and its isoenzymes in urine methods and clinical applications [J].Biochemistry Section,1992 ,30(10):693~705.
[14] NIPPON RINSHO. N-acetyl-beta-D-glucosaminidase.1995 ,53(5):1267~76
[15] Murayama M,Hiramatsu M,Kashimata M,et al. Properties of N-acetyl-beta-glucosaminidase in the masseter muscle of the mouse[J].Shika Kiso Igakkai Zasshi,1990 ,32(1):1~9
[16] OOSTHUIZEN MM,LAMBRECHTS H,MYBURGH JA,et al.Properties of the isoenzyme forms A-1,A-2 and B of N-acetyl-beta-D-glucosaminidase purified from baboon kidneys Beukes HA[J]. Int J Biochem.1991,23(2):239~51
[17]黄小红,罗忠宝,等.猪精液碱性N-乙酰-β-D-氨基葡萄糖苷酶的纯化与酶学性质研究[J].中国农业科学,2008,41(9):2843~2849
[18] BERNARD WEISSMANN,SEMELE HADJIIOANNOU,JERALD TORNHEIM.Oligosaccharase Activity ofβ-N-Acetyl-D-glucosaminidase of beef liver[J].Biological Chemistry,1964,239(1):59~63
[19]JACOB A.VEHPOORTE.Purification of Twoβ-N-Acetyl-D-glucosaminidase from Beef Spleen [J].Biological Chemistry,1972,247(15):4781~4793
[20] JAUHIAINEN A, VANHA-PERTTULA T. beta-N-acetylglucosaminidase in the reproductive organs and seminal plasma of the bull[J].J Reprod Fertil,1986 ,76(1):239~50
[21] JAUHIAINEN A, VANHA-PERTTULA T. Beta-galactosidase in the seminal plasma and reproductive organs of the bull [J],Int J Biochem ,1986,18(8):719~24.
[22] DROBA B, DROBA M. Properties of acid beta-N-acetyl-D-glucosaminidase from cock semen [J].Folia Biol (Krakow),1992,40(12):67~71.
[23]王长康,黄一帆,黄小红,等.番鸭睾丸N-乙酰-β-D-氨基葡萄糖苷酶的提取及性质研究[J].畜牧兽医学报,2007,38(6):552~557
[24]王长康,艾启青,黄小红,等.山麻鸭睾丸和肝脏中N-乙酰-β-D-氨基葡萄糖苷酶的分离提取及其性质的比较[J].福建农林大学学报(自然科学版),2009,38(1):49~54
[25]黄小红,金艳东,等.河田鸡与艾维茵肉鸡睾丸N-乙酰-β-D-氨基葡萄糖苷酶的特性比较[J].福建农林大学学报(自然科学版),2009,38(2):49~54.
[26]程明哲,苏拔贤,宋英良等.虾头几丁质酶的研究[J].热带海洋,1996,15(2): 68~73.
[27]李友宾.罗氏沼虾肝胰腺几丁质酶的亲和消化纯化方法初探[J].湛江海洋大学学报,2001,21(2):69~71.
[28]KONO M ,MATSUI T,SHIMIZU C,et a1.Purifications and some properties ofchitinase from the liver of aprawn,Penaeus japonicus[J].Agric Biol Chem,1990,54(8):2145~2147.
[29] LYNN K R.Chitinases and Chitobioases from the American Lobster Homartts[J]. Biochem Physiol,1990,96B :76~766.
[30]KOGA D,NAKASHIMA M,MATSUKURA T,et al.Purification and properties ofβ-N-acetyl-D-glucosaminidase from alimentary canal of the silkworm,Bombyx mori[J].Agric.Biol.Chem,1986,5(9):2357~2368.
[31]陈宏惠.效应物对中华绒螯蟹N-乙酰-β-D-氨基葡萄糖苷酶的影响[D],福州:福建农林大学,2007.
[32] ESPIE P J,ROFF J C.Characterization of chitobiase from Daphnia magna and its relation to chitin flux[J].Physiol Zool,1995,68:727~748.
[33]SPINDLER BARTH M,VAN WORMHOUDT A,SPINDLER KD.Chitinolytic enzymes in the integument and midgut .gland of the shrimp Palaemon serratus during the moulting cycle[J].Mar Biol,1990,106:49~52.
[34]SOMMER U.Physical properties of beta-N-acetyl-D-glucosaminidase and beta-N-acetyl-D-hexosaminidase from Drosophila Kc-cells[J]. Arch Insect Biochem Physiol ,1991;18(1):45~53.
[35]SOMMER U.Demonstration of beta-N-acetyl-D-glucosaminidase and beta-N-acetyl-D-hexosaminidase in Drosophila Kc-cells[J].Arch Insect Biochem Physiol. 1991;17(1):3~13
[36]BEDI GS, SHAH RH,BAHL OP.Studies on Turbatrix aceti beta-N-acetylglucosaminidase. 1. Purification and physicochemical characterization, 1984 , 233(1):237~50.
[37] JIN YL, JO YY,KIM KY,et al.Purification and characterization of beta-N-acetylhexosaminidase from rice seeds[J]. Department of Agricultural Chemistry and Institute of Agricultural Science and Technology,2008,4:500~757.
[38]史益敏,颜季琼,费雪南,等.番茄叶片胞外β-N-乙酰氨基己糖苷酶的部分性质研究[J].植物学报,1994,36(11):864~870
[39]史益敏,颜季琼,费雪南,等.从感染TMV的番茄叶纯化胞外β-N-乙酰氨基己糖苷酶[J].植物学报,1994,36 (10) : 773~778
[40]SCHLUMBAUM A,MAUCH F,VOGELI U,et al.Plant chitinase are potent inhibitors of fungal growth[J].Nature,1986,324:355~356.
[41]李奕雅,陈凡,等.花生几丁质酶的纯化及酶学性质研究[J].漳州师范学院学报(自然科学版) ,2008(3):79~82.
[42]曾艳,赵南明,刘进元.几丁质酶与植物防卫反应[J].生物工程进展,1997,17(4):31~34
[43]杨革,陈洪章,李佐虎.绿僵菌几丁质酶的分离纯化及性质[J].化工学报.2005,56(4):673~676.
[44]王治伟,刘志敏.微生物几丁质酶研究进展[J].生物技术通讯.2006(3):439~442.
[45] MOJTABA M.DAN FUNCK J. An N-acetyl-b-D-glucosaminidase gene, cr-nag1, from thebiocontrol agent Clonostachys rosea is up-regulated in antagonistic interactions with Fusarium culmorum[M].Mycological Research,2009:33~43.
[46] KRAMER K J,CORPUS L M,CHOI H,et a1. Sequence of cDNA and expression of the gene encoding epidermal and gut chitinases of Manduca Sexta[J].Insect Biochem Mol Biol,1993,23:691~701.
[47]Y-P ZHENGP,J.KRELL D.Cloning,Expression,and Localization of a Molt Relatedβ-N-Acetylglucosaminidase in the Spruce Budworm,Choristoneura fumiferna.Archives of Insect Biochemistry and Physiology,2008( 68):49~50.
[48] AHMAD T, RAJAGOPAL R,BHATNAGAR R. K. Biochem.Biophys . Res . Commun,2003,310 (1) :188～195.
[49]李华钟,王树英,等. Clostridium paraputrificumM-21β-N-乙酰氨基葡萄糖苷酶的表达、纯化与性质[J].无锡轻工大学学报,2003,22(3):41~45.
[50]]SAKUDA S,ISOGAI A,MATSUMOTO S,et a1.Search formicrobial insect growth regulators.II. Allosamidin.anovel insect chltinase inhibitor[J].J Antibiot,1987,40(3):296
[51]周泽扬,作田壮平,山田靖宙,等.几丁质酶抑制物allosamidin中所含天然N一乙酰基一D氨基阿洛糖的生成机制探讨[J].中国抗生索杂志,1996,21(4):298.
[52]ZHOU Z Y,SAKUDA S,KINOSHITA M,et a1.Biosyntheticstudies of allosamidin. 2.Isolation of didemethylallosamidin,and conversion experiments of‘14C—labeled demethylallosamidin’, didemethylallosamidin and their related compounds[J].J Antibiot,1993,46(10):1582.
[53]ZHOU Z Y,SAKUDA S,YAMADA Y.Biosynthetic studies on the chitinase inhibitor,allosamidin.Origin of the carbon and nitrogen atoms[J].J Chem Soc Perkin Tran 1,1992,13:1649
[54]MERZENDOFER H .ZIMOCH L .Chitin metabolism in insects:structure,function and regulation of chitin synthases and chitinases[J].J Exp Biol,2003.206:4393
[55]SAKUDA S,NISHIMOTO Y,OHI M,et a1.Effects of demethyallosamidin,a potent yeast chitinase inhibitor,on the cell division of yeast[J].Agr Biol Chem,1990,54(5):1333.
[56] HOUSTON D R,EGGLESTON I,SYNSTAD B,et a1.The cyclicdipeptide CI-4[cyclo一(L—Arg—D—Pro)]inhibits family 18 chitinases by structural mimicry of a reaction inter mediate [J].Biochem J,2002.368(1):23.
[57] HOUSTON D R,SHIOMI K,ARAI N.et a1.High resolution structures of a chitinase complexed with natural product cyclopentapeptide inhibitors: Mimicry of carbohydratesubstrate[J].PNAS,2002,99(14):9127.
[58]ZHU Z Y,ZHENG T,HOMER R J,et a1.Acidic mammalian chitinase in asthmatic Th2 inflammation and IL-13 pathway activation[J].Science,2004,304(5677)l 1678
[59]潘晓芳,谢晓兰,徐海金,赖志青.有机溶剂对黄粉虫N-乙酰-β-D-氨基葡萄糖苷酶活力影响[J].宁德师专学报(自然科学版),2006(4):352~355.
[60]林建成,谢晓兰,龚敏.有机溶剂对南美白对虾N-乙酰-β-D-氨基葡萄糖苷酶活力的影响[J].厦门大学学报(自然科学版),2005(5):693~696.
[61]谢进金,张继平,王勤,杨学敏,林建成,陈清西锯缘青蟹NAGase在甲醛溶液中的失活动力学研究[J].厦门大学学报(自然科学版),2006年(1):98~101
[62] XIE X L,CHEN Q X.Inactivation kinetics ofβ-N-Acetyl-D-glucosaminidase from prawn(Penaeus vannamei)in the dioxane solution[J].Biochemistry Moscow,2004,69(12):1365~1371.
[63]CHEN Q X,ZHANG Z,HUANG H,et al.Unfolding and inactivation of Ampullarrium crosseanβ-glucosidase during denaturation by guanidinium hydrochloride[J].Int.J.Biochem.Cell Biol,2003,35:1227~1233.
[64]MOUDNI B E,RODIER M H,JACQUEMIN J L.Purification and characterization of N-acetyl glucosaminidase from Trypanosoma cruzi[J].Experimental Parasitology,1996,83:167~173.
[65]杨学敏,谢进金,张继平,等.金属离子对锯缘青蟹NAGase活力的影响[J].台湾海峡,2006,25(3):343~346.
[66]颜雅雯,张继平,等.抗菌类药物对锯缘青蟹N-乙酰-β-D-氨基葡萄糖苷酶活力的影响[J].台湾海峡.2007(26),2:231~235.
[67] LIN J C,CHEN Q X,SHI Y,et al.The chemical modification of the essential groups of beta-N-acetyl-D-glucosaminidase from Turbo cornutus solander[J].IUBMB Life,2003,55(9):547~552.
[68]张继平.锯缘青蟹N-乙酰-beta-D-氨基葡萄糖苷酶的性质及活力调控的研究[D].厦门:厦门大学,2006.
[69]谢晓兰.南美白对虾N-乙酰-beta-D-氨基葡萄糖苷酶的研究[D].厦门大学,2005.
[70] ZEN K C,CHOI H K,KRISHNAMACHARY N,et al. Cloning,expression,and hormonal regulation of an insect(-N-acety-glucosaminidase gene[J].Insect Biochem.Mol Biol,1996,26(5):435~444
[71]石艳,刘炜风,陈清西.菜青虫NAGase活性必需集团的研究[J].厦门大学学报(自然科学版),2006,45(6):851~853 .
[72] BARBERM SKR IDE J P. Purification and p roperties of a wheal leaf N–acetyl-β-D hexosaminidase[J]. P lant S ciK,1989.,60:163~172.
[73] TETSURO SHINODA , et al.Cloning and functional expression of a chitinase cDNA from the comon cutworm,Spodoptera litura ,using a recombinant baculovirus lacking the virus - encoded chitinase gene [J ].Insect Biochemistry and Molecular Biology,2001 (31) :521~532.
[74]张九斌. N-乙酰-β-D-氨基葡萄糖苷酶(NAG)测定及其临床意义[J].齐鲁医学检验,2004,15(2):62.
[75]张谙,王诗瑾.精液NAGase活力的测定及其临床应用[J].湖南医学,1997(14):47.
[76]舒新亚,龚珞军.淡水小龙虾健康养殖实用技术[M].中国农业出版社.
[77]Fingerman M.Crustacean endocrinology:a retrospective, prospective,and introspective analysis[J].Physiol Zoo1.1997,70:257~269.
[78]LOWRY O H,ROSEBROUGH N J,FARR A L,et al.Protein measurement with the Folin phenol reagent [J].J.Biol.Chem.,1951,193(1):265~275
[79]KONO M,MATSUI T,SHIMIZU C,et al.Purifications and some properties of chitinase from the liver of a Prawn,Penaeus japonicus[J].Agric.Biol.Chem.,1990,54(8):2145~2147
[80]CHEN Q X,ZHANG W,ZHENG W Z,et al.Kinetics of inhibition of alkaline phosphatase from green crab(Scylla serrata)by N-bromosuccinimide[J].J.Protein Chem.,1996,15:345~350
[81]陈清西.锯缘青蟹碱性磷酸酶结构与功能的研究[D].北京:清华大学,1999
[82]李建武,萧能赓,余瑞元,等.生物化学实验原理和方法[M].北京:北京大学出版社,2002
[83]陆建.蛋白质纯化技术及应用[M].化学工业出版社,2005
[84] ARNOULD C,JEUNIAUX C.Cah Biol,Les enzymes hydrolytiques du. système digestif chez les crustacés pagurides. Cah. Biol. Mar. 1982,23:89-103
[85] ESPIE P J,ROFF J C.Characterization of chitobiase from Daphnia magna and its relation to chitin flux[J].Physiol Zool,1995,68:727~748.
[86]SPINDLER BARTH M,VAN WORMHOUDT A,SPINDLER KD.Chitinolytic enzymes in the integument and midgut .gland of the shrimp Palaemon serratus during the moulting cycle[J].Mar Biol,1990,106:49~52.
[87]李友宾.罗氏沼虾肝胰腺几丁质酶分离纯化及部分性质[J].西南民族大学学报(自然科学版):2001.3
[88] Sakai T, Nakanishi Y, Kato I. Purification and characterization of beta-N-acetyl-D-hexosaminidase from the mid-gut gland of scallop (Patinopecten yessoensis) [J].Biosci Biotechnol Biochem. 1993 Jun;57(6):965~8.
[89] Shigeta S, Matsuda A, Oka S. J Biochem..Purification and characterization of a beta-N-acetylhexosaminidase of sea-squirt .1982 Jul;92(1):163~72.
[90] DROBA B, DROBA M. Properties of acid beta-N-acetyl-D-glucosaminidase from cocksemen [J].Folia Biol (Krakow),1992,40(1-2):67~71.
[91]黄小红,王寿昆,等.日本沼虾N-乙酰-β-D-氨基葡萄糖苷酶初步纯化及部分性质[J].应用与环境生物学报. 2006,12(6): 804~80.
[92]林建城,王悦,谢晓兰,等.金属离子对凡纳对虾N-乙酰-β-D-氨基葡萄糖苷酶活力的影响[J].台湾海峡,2005,24(1):78~82.
[93]Chen Q X,Zheng W Z,Lin J Y ,et al.Effect of metal ions on the activitu of green crab (Scylla serrata ) alkaline phosphatase[J]. Int J Biochem Cell Biol ,2000,32(8):879~885.
[94]Kenji Y ,K wang M L,Hidehiko K,ea tl.Purification and characterization ofβ-N-acetulhexosaminidase from Penicilium oxalicum[J].Agric Biol Chem,1985,49(3):611~619 .
[95]Koga D,Nakashima M,Matsukura T,et al.Purifications and characterization ofβ-N-acetyl-D-glucosaminidase from alimentaru canal of the silkworm,Bombyx mori[J]. Agric Biol Chem,1986,50(9):2357~2368.
[96]谢晓兰,王悦,陈清西.氨基酸对凡纳滨对虾N-乙酰-β-D-氨基葡萄糖苷酶活力的影响[J].海洋科学,2006,30(3):46~50.
[97]林建成,谢进金,张继平,等.氨基酸对锯缘青蟹N-乙酰氨基葡萄糖苷酶的效应[J].厦门大学学报(自然科学版),2006,45(2):264~267.
[98]黄乾生,谢晓兰,等.不同生长期凡纳滨对虾肝胰腺NAGase的基本性质变化[J].厦门大学学报(自然科学版)2006,45(6):847~849
[2]KEYHANI N O,WANG L X,LEE Y C.The chitin catabolic cascade in the marine bacterium Vibrio furnissii-Characterization of an N,N’-diacetyl-chitobiose transport system[J].Journal of Biological Chemistry,1996,271(52):33409~33413.
[3]BOLLER J,GEHRI A,MAUCH F.Chitinase in bean leaves:induction by ethylene,purification,properties,and possible function[J].Planta,1983,157:22~31.
[4]WERRIES E,NEUE I,BUDDECKE E.Evidence for different glycohydrolase and glycosyltransferase activities of beta-N-acetylglucosaminidases A and B[J].Hoppe Seylers Z Physiol Chem,1975,356(6):953~960.
[5]KONDO K,MATSUMOTO M,KOJO A,et al.Purification and characterization of chitinase from pupae Pieris rapae crucivora(Boiduval)[J].Journal of Chemical and Engineering of Japan,2002,35:241~246.
[6]USUI T,HAYASHI Y,NANJO F,et al.Transglycosylation reaction of a chitinase purified from Nocardia orientalis[J].Biochim Biophys Acta,1987,923(8):302~309.
[7]TAKEGAWA K,FUJITA K,FAN J Q,et al..Enzymatic synthesis of a neoglycoconjugate by transglycosylation with Arthrobacter endo-β-N-acetylglucosaminidase:A substrate for colorimetric detection of endo-β-N-acetylglucosaminidase activity.Analytical Biochemistry[J].1998,257(2):218~223.
[8] LIN JIAN CHENG, CHEN Q X, SHI Y,et al. The chemical modification of the essential groups ofβ-N-Acetyl-D-glucosaminidase from Turbo cornutus Solander[J]. IUBMB Life,2003,55 (9): 547~552.
[9] ZOU E,FINGERMAN M. Chitobiase activity in the epidermis and hepatopancreas of the fiddler crab Uca pugilator during the molting cycle[J]. Marine Biology,1999,133: 97~101.
[10]高磊,蔡国平.小鼠几丁质酶的重组表达和抗体制备[J].细胞与分子免疫学杂志,2008 9(25):9
[11]YOSHIDA K,UCHIJIMA Y, KOBAYASHI N,et al. Separation and properties of N-acetyl-beta-D-glucosaminidase from human seminal plasma . Article in Japanese[J] Nippon Hinyokika Gakkai Zasshi.1989 Nov;80(11):1625~31.
[12]YOSHIDA K,TAKAHASHI T,NAKAME Y ,et al.N-acetyl-beta-D-glucosaminidase (NAG) activity in human semen: its relation to gamma-glutamyl transpeptidase (gamma-GTP) activity in seminal plasma and reproductive tissues,and relation between seminal mucoprotein concentration and seminal NAG,and gamma-GTP activities[J]. Hinyokika Kiyo.1987 Jul;33(7):1054~9.
[13] EUR J CLIN CHEM CLIN BIOCHEM. Measurement of N-acetyl-beta-glucosaminidase and its isoenzymes in urine methods and clinical applications [J].Biochemistry Section,1992 ,30(10):693~705.
[14] NIPPON RINSHO. N-acetyl-beta-D-glucosaminidase.1995 ,53(5):1267~76
[15] Murayama M,Hiramatsu M,Kashimata M,et al. Properties of N-acetyl-beta-glucosaminidase in the masseter muscle of the mouse[J].Shika Kiso Igakkai Zasshi,1990 ,32(1):1~9
[16] OOSTHUIZEN MM,LAMBRECHTS H,MYBURGH JA,et al.Properties of the isoenzyme forms A-1,A-2 and B of N-acetyl-beta-D-glucosaminidase purified from baboon kidneys Beukes HA[J]. Int J Biochem.1991,23(2):239~51
[17]黄小红,罗忠宝,等.猪精液碱性N-乙酰-β-D-氨基葡萄糖苷酶的纯化与酶学性质研究[J].中国农业科学,2008,41(9):2843~2849
[18] BERNARD WEISSMANN,SEMELE HADJIIOANNOU,JERALD TORNHEIM.Oligosaccharase Activity ofβ-N-Acetyl-D-glucosaminidase of beef liver[J].Biological Chemistry,1964,239(1):59~63
[19]JACOB A.VEHPOORTE.Purification of Twoβ-N-Acetyl-D-glucosaminidase from Beef Spleen [J].Biological Chemistry,1972,247(15):4781~4793
[20] JAUHIAINEN A, VANHA-PERTTULA T. beta-N-acetylglucosaminidase in the reproductive organs and seminal plasma of the bull[J].J Reprod Fertil,1986 ,76(1):239~50
[21] JAUHIAINEN A, VANHA-PERTTULA T. Beta-galactosidase in the seminal plasma and reproductive organs of the bull [J],Int J Biochem ,1986,18(8):719~24.
[22] DROBA B, DROBA M. Properties of acid beta-N-acetyl-D-glucosaminidase from cock semen [J].Folia Biol (Krakow),1992,40(12):67~71.
[23]王长康,黄一帆,黄小红,等.番鸭睾丸N-乙酰-β-D-氨基葡萄糖苷酶的提取及性质研究[J].畜牧兽医学报,2007,38(6):552~557
[24]王长康,艾启青,黄小红,等.山麻鸭睾丸和肝脏中N-乙酰-β-D-氨基葡萄糖苷酶的分离提取及其性质的比较[J].福建农林大学学报(自然科学版),2009,38(1):49~54
[25]黄小红,金艳东,等.河田鸡与艾维茵肉鸡睾丸N-乙酰-β-D-氨基葡萄糖苷酶的特性比较[J].福建农林大学学报(自然科学版),2009,38(2):49~54.
[26]程明哲,苏拔贤,宋英良等.虾头几丁质酶的研究[J].热带海洋,1996,15(2): 68~73.
[27]李友宾.罗氏沼虾肝胰腺几丁质酶的亲和消化纯化方法初探[J].湛江海洋大学学报,2001,21(2):69~71.
[28]KONO M ,MATSUI T,SHIMIZU C,et a1.Purifications and some properties ofchitinase from the liver of aprawn,Penaeus japonicus[J].Agric Biol Chem,1990,54(8):2145~2147.
[29] LYNN K R.Chitinases and Chitobioases from the American Lobster Homartts[J]. Biochem Physiol,1990,96B :76~766.
[30]KOGA D,NAKASHIMA M,MATSUKURA T,et al.Purification and properties ofβ-N-acetyl-D-glucosaminidase from alimentary canal of the silkworm,Bombyx mori[J].Agric.Biol.Chem,1986,5(9):2357~2368.
[31]陈宏惠.效应物对中华绒螯蟹N-乙酰-β-D-氨基葡萄糖苷酶的影响[D],福州:福建农林大学,2007.
[32] ESPIE P J,ROFF J C.Characterization of chitobiase from Daphnia magna and its relation to chitin flux[J].Physiol Zool,1995,68:727~748.
[33]SPINDLER BARTH M,VAN WORMHOUDT A,SPINDLER KD.Chitinolytic enzymes in the integument and midgut .gland of the shrimp Palaemon serratus during the moulting cycle[J].Mar Biol,1990,106:49~52.
[34]SOMMER U.Physical properties of beta-N-acetyl-D-glucosaminidase and beta-N-acetyl-D-hexosaminidase from Drosophila Kc-cells[J]. Arch Insect Biochem Physiol ,1991;18(1):45~53.
[35]SOMMER U.Demonstration of beta-N-acetyl-D-glucosaminidase and beta-N-acetyl-D-hexosaminidase in Drosophila Kc-cells[J].Arch Insect Biochem Physiol. 1991;17(1):3~13
[36]BEDI GS, SHAH RH,BAHL OP.Studies on Turbatrix aceti beta-N-acetylglucosaminidase. 1. Purification and physicochemical characterization, 1984 , 233(1):237~50.
[37] JIN YL, JO YY,KIM KY,et al.Purification and characterization of beta-N-acetylhexosaminidase from rice seeds[J]. Department of Agricultural Chemistry and Institute of Agricultural Science and Technology,2008,4:500~757.
[38]史益敏,颜季琼,费雪南,等.番茄叶片胞外β-N-乙酰氨基己糖苷酶的部分性质研究[J].植物学报,1994,36(11):864~870
[39]史益敏,颜季琼,费雪南,等.从感染TMV的番茄叶纯化胞外β-N-乙酰氨基己糖苷酶[J].植物学报,1994,36 (10) : 773~778
[40]SCHLUMBAUM A,MAUCH F,VOGELI U,et al.Plant chitinase are potent inhibitors of fungal growth[J].Nature,1986,324:355~356.
[41]李奕雅,陈凡,等.花生几丁质酶的纯化及酶学性质研究[J].漳州师范学院学报(自然科学版) ,2008(3):79~82.
[42]曾艳,赵南明,刘进元.几丁质酶与植物防卫反应[J].生物工程进展,1997,17(4):31~34
[43]杨革,陈洪章,李佐虎.绿僵菌几丁质酶的分离纯化及性质[J].化工学报.2005,56(4):673~676.
[44]王治伟,刘志敏.微生物几丁质酶研究进展[J].生物技术通讯.2006(3):439~442.
[45] MOJTABA M.DAN FUNCK J. An N-acetyl-b-D-glucosaminidase gene, cr-nag1, from thebiocontrol agent Clonostachys rosea is up-regulated in antagonistic interactions with Fusarium culmorum[M].Mycological Research,2009:33~43.
[46] KRAMER K J,CORPUS L M,CHOI H,et a1. Sequence of cDNA and expression of the gene encoding epidermal and gut chitinases of Manduca Sexta[J].Insect Biochem Mol Biol,1993,23:691~701.
[47]Y-P ZHENGP,J.KRELL D.Cloning,Expression,and Localization of a Molt Relatedβ-N-Acetylglucosaminidase in the Spruce Budworm,Choristoneura fumiferna.Archives of Insect Biochemistry and Physiology,2008( 68):49~50.
[48] AHMAD T, RAJAGOPAL R,BHATNAGAR R. K. Biochem.Biophys . Res . Commun,2003,310 (1) :188～195.
[49]李华钟,王树英,等. Clostridium paraputrificumM-21β-N-乙酰氨基葡萄糖苷酶的表达、纯化与性质[J].无锡轻工大学学报,2003,22(3):41~45.
[50]]SAKUDA S,ISOGAI A,MATSUMOTO S,et a1.Search formicrobial insect growth regulators.II. Allosamidin.anovel insect chltinase inhibitor[J].J Antibiot,1987,40(3):296
[51]周泽扬,作田壮平,山田靖宙,等.几丁质酶抑制物allosamidin中所含天然N一乙酰基一D氨基阿洛糖的生成机制探讨[J].中国抗生索杂志,1996,21(4):298.
[52]ZHOU Z Y,SAKUDA S,KINOSHITA M,et a1.Biosyntheticstudies of allosamidin. 2.Isolation of didemethylallosamidin,and conversion experiments of‘14C—labeled demethylallosamidin’, didemethylallosamidin and their related compounds[J].J Antibiot,1993,46(10):1582.
[53]ZHOU Z Y,SAKUDA S,YAMADA Y.Biosynthetic studies on the chitinase inhibitor,allosamidin.Origin of the carbon and nitrogen atoms[J].J Chem Soc Perkin Tran 1,1992,13:1649
[54]MERZENDOFER H .ZIMOCH L .Chitin metabolism in insects:structure,function and regulation of chitin synthases and chitinases[J].J Exp Biol,2003.206:4393
[55]SAKUDA S,NISHIMOTO Y,OHI M,et a1.Effects of demethyallosamidin,a potent yeast chitinase inhibitor,on the cell division of yeast[J].Agr Biol Chem,1990,54(5):1333.
[56] HOUSTON D R,EGGLESTON I,SYNSTAD B,et a1.The cyclicdipeptide CI-4[cyclo一(L—Arg—D—Pro)]inhibits family 18 chitinases by structural mimicry of a reaction inter mediate [J].Biochem J,2002.368(1):23.
[57] HOUSTON D R,SHIOMI K,ARAI N.et a1.High resolution structures of a chitinase complexed with natural product cyclopentapeptide inhibitors: Mimicry of carbohydratesubstrate[J].PNAS,2002,99(14):9127.
[58]ZHU Z Y,ZHENG T,HOMER R J,et a1.Acidic mammalian chitinase in asthmatic Th2 inflammation and IL-13 pathway activation[J].Science,2004,304(5677)l 1678
[59]潘晓芳,谢晓兰,徐海金,赖志青.有机溶剂对黄粉虫N-乙酰-β-D-氨基葡萄糖苷酶活力影响[J].宁德师专学报(自然科学版),2006(4):352~355.
[60]林建成,谢晓兰,龚敏.有机溶剂对南美白对虾N-乙酰-β-D-氨基葡萄糖苷酶活力的影响[J].厦门大学学报(自然科学版),2005(5):693~696.
[61]谢进金,张继平,王勤,杨学敏,林建成,陈清西锯缘青蟹NAGase在甲醛溶液中的失活动力学研究[J].厦门大学学报(自然科学版),2006年(1):98~101
[62] XIE X L,CHEN Q X.Inactivation kinetics ofβ-N-Acetyl-D-glucosaminidase from prawn(Penaeus vannamei)in the dioxane solution[J].Biochemistry Moscow,2004,69(12):1365~1371.
[63]CHEN Q X,ZHANG Z,HUANG H,et al.Unfolding and inactivation of Ampullarrium crosseanβ-glucosidase during denaturation by guanidinium hydrochloride[J].Int.J.Biochem.Cell Biol,2003,35:1227~1233.
[64]MOUDNI B E,RODIER M H,JACQUEMIN J L.Purification and characterization of N-acetyl glucosaminidase from Trypanosoma cruzi[J].Experimental Parasitology,1996,83:167~173.
[65]杨学敏,谢进金,张继平,等.金属离子对锯缘青蟹NAGase活力的影响[J].台湾海峡,2006,25(3):343~346.
[66]颜雅雯,张继平,等.抗菌类药物对锯缘青蟹N-乙酰-β-D-氨基葡萄糖苷酶活力的影响[J].台湾海峡.2007(26),2:231~235.
[67] LIN J C,CHEN Q X,SHI Y,et al.The chemical modification of the essential groups of beta-N-acetyl-D-glucosaminidase from Turbo cornutus solander[J].IUBMB Life,2003,55(9):547~552.
[68]张继平.锯缘青蟹N-乙酰-beta-D-氨基葡萄糖苷酶的性质及活力调控的研究[D].厦门:厦门大学,2006.
[69]谢晓兰.南美白对虾N-乙酰-beta-D-氨基葡萄糖苷酶的研究[D].厦门大学,2005.
[70] ZEN K C,CHOI H K,KRISHNAMACHARY N,et al. Cloning,expression,and hormonal regulation of an insect(-N-acety-glucosaminidase gene[J].Insect Biochem.Mol Biol,1996,26(5):435~444
[71]石艳,刘炜风,陈清西.菜青虫NAGase活性必需集团的研究[J].厦门大学学报(自然科学版),2006,45(6):851~853 .
[72] BARBERM SKR IDE J P. Purification and p roperties of a wheal leaf N–acetyl-β-D hexosaminidase[J]. P lant S ciK,1989.,60:163~172.
[73] TETSURO SHINODA , et al.Cloning and functional expression of a chitinase cDNA from the comon cutworm,Spodoptera litura ,using a recombinant baculovirus lacking the virus - encoded chitinase gene [J ].Insect Biochemistry and Molecular Biology,2001 (31) :521~532.
[74]张九斌. N-乙酰-β-D-氨基葡萄糖苷酶(NAG)测定及其临床意义[J].齐鲁医学检验,2004,15(2):62.
[75]张谙,王诗瑾.精液NAGase活力的测定及其临床应用[J].湖南医学,1997(14):47.
[76]舒新亚,龚珞军.淡水小龙虾健康养殖实用技术[M].中国农业出版社.
[77]Fingerman M.Crustacean endocrinology:a retrospective, prospective,and introspective analysis[J].Physiol Zoo1.1997,70:257~269.
[78]LOWRY O H,ROSEBROUGH N J,FARR A L,et al.Protein measurement with the Folin phenol reagent [J].J.Biol.Chem.,1951,193(1):265~275
[79]KONO M,MATSUI T,SHIMIZU C,et al.Purifications and some properties of chitinase from the liver of a Prawn,Penaeus japonicus[J].Agric.Biol.Chem.,1990,54(8):2145~2147
[80]CHEN Q X,ZHANG W,ZHENG W Z,et al.Kinetics of inhibition of alkaline phosphatase from green crab(Scylla serrata)by N-bromosuccinimide[J].J.Protein Chem.,1996,15:345~350
[81]陈清西.锯缘青蟹碱性磷酸酶结构与功能的研究[D].北京:清华大学,1999
[82]李建武,萧能赓,余瑞元,等.生物化学实验原理和方法[M].北京:北京大学出版社,2002
[83]陆建.蛋白质纯化技术及应用[M].化学工业出版社,2005
[84] ARNOULD C,JEUNIAUX C.Cah Biol,Les enzymes hydrolytiques du. système digestif chez les crustacés pagurides. Cah. Biol. Mar. 1982,23:89-103
[85] ESPIE P J,ROFF J C.Characterization of chitobiase from Daphnia magna and its relation to chitin flux[J].Physiol Zool,1995,68:727~748.
[86]SPINDLER BARTH M,VAN WORMHOUDT A,SPINDLER KD.Chitinolytic enzymes in the integument and midgut .gland of the shrimp Palaemon serratus during the moulting cycle[J].Mar Biol,1990,106:49~52.
[87]李友宾.罗氏沼虾肝胰腺几丁质酶分离纯化及部分性质[J].西南民族大学学报(自然科学版):2001.3
[88] Sakai T, Nakanishi Y, Kato I. Purification and characterization of beta-N-acetyl-D-hexosaminidase from the mid-gut gland of scallop (Patinopecten yessoensis) [J].Biosci Biotechnol Biochem. 1993 Jun;57(6):965~8.
[89] Shigeta S, Matsuda A, Oka S. J Biochem..Purification and characterization of a beta-N-acetylhexosaminidase of sea-squirt .1982 Jul;92(1):163~72.
[90] DROBA B, DROBA M. Properties of acid beta-N-acetyl-D-glucosaminidase from cocksemen [J].Folia Biol (Krakow),1992,40(1-2):67~71.
[91]黄小红,王寿昆,等.日本沼虾N-乙酰-β-D-氨基葡萄糖苷酶初步纯化及部分性质[J].应用与环境生物学报. 2006,12(6): 804~80.
[92]林建城,王悦,谢晓兰,等.金属离子对凡纳对虾N-乙酰-β-D-氨基葡萄糖苷酶活力的影响[J].台湾海峡,2005,24(1):78~82.
[93]Chen Q X,Zheng W Z,Lin J Y ,et al.Effect of metal ions on the activitu of green crab (Scylla serrata ) alkaline phosphatase[J]. Int J Biochem Cell Biol ,2000,32(8):879~885.
[94]Kenji Y ,K wang M L,Hidehiko K,ea tl.Purification and characterization ofβ-N-acetulhexosaminidase from Penicilium oxalicum[J].Agric Biol Chem,1985,49(3):611~619 .
[95]Koga D,Nakashima M,Matsukura T,et al.Purifications and characterization ofβ-N-acetyl-D-glucosaminidase from alimentaru canal of the silkworm,Bombyx mori[J]. Agric Biol Chem,1986,50(9):2357~2368.
[96]谢晓兰,王悦,陈清西.氨基酸对凡纳滨对虾N-乙酰-β-D-氨基葡萄糖苷酶活力的影响[J].海洋科学,2006,30(3):46~50.
[97]林建成,谢进金,张继平,等.氨基酸对锯缘青蟹N-乙酰氨基葡萄糖苷酶的效应[J].厦门大学学报(自然科学版),2006,45(2):264~267.
[98]黄乾生,谢晓兰,等.不同生长期凡纳滨对虾肝胰腺NAGase的基本性质变化[J].厦门大学学报(自然科学版)2006,45(6):847~849