卢慧茵,王炜,崔春

• 1:华南理工大学食品科学与工程学院

摘要(Abstract)：

以2种常见的谷氨酰胺酶作为研究对象,利用离子选择电极法测定谷氨酰胺酶的水解活性和转肽活性,探究不同pH、温度、盐含量对谷氨酰胺酶的水解活性及转肽活性的影响。研究结果表明:厂家一谷氨酰胺酶催化水解反应的最适pH为7.0,最适温度为50℃,催化转肽反应的最适pH为10.0,最适温度为50℃;厂家二谷氨酰胺酶催化水解反应的最适pH为7.0,最适温度为45℃,催化转肽反应的最适pH为9.0,最适温度为45℃。同时,谷氨酰胺酶表现水解活性时具有较好的耐盐性,但其转肽活性随盐含量的增加而下降。该研究结果可为谷氨酰胺酶的产业化应用提供理论指导。

关键词(KeyWords)： 谷氨酰胺酶;影响因素;水解;转肽;酶活

Abstract:

Keywords:

基金项目(Foundation): 中央高校基本业务费项目(2017ZD093);; 广东省科技计划项目(2017A010105002)

作者(Author): 卢慧茵,王炜,崔春

参考文献(References)：

• [1]詹寿年,吴拥军,郭倩倩,等.高活性谷氨酰胺酶豆豉芽孢杆菌的筛选及glsA基因的克隆[J].贵州农业科学,2011,39(6):119-122.
• [2]Sato I,Kobayashi H,Hanya Y,et al.Cryptococcus nodaensis sp.nov.,ayeast isolated from soil in Japan that produces a salt-tolerant and thermostable glutaminase[J].Journal of Industrial Microbiology&Biotechnology,1999,22(3):127-132.
• [3]何灿,高红亮.谷氨酰胺酶的应用研究进展[J].安徽农业科学,2012,40(32):15883-15885.
• [4]Singh P,Shera S S,Banik J,et al.Optimization of cultural conditions using response surface methodology versus artificial neural network and modeling of L-glutaminase production by Bacillus cereus MTCC 1305[J].Bioresource Technology,2013,137:261-269.
• [5]Lebedeva Z I,Kabanova E A,Berezov T T.The nature of functional groups in the active center of antitumor glutamin-(asparagin-)ase[J].Bulletin of Experimental Biology&Medicine,1988,105(4):507-510.
• [6]El-Sayed A S A.L-glutaminase production by Trichoderma koningii,under solid-state fermentation[J].Indian Journal of Microbiology,2009,49(3):243-250.
• [7]Aryuman P,Lertsiri S,Visessanguan W,et al.Glutaminaseproducing Meyerozyma(Pichia)guilliermondii isolated from Thai soy sauce fermentation[J].International Journal of Food Microbiology,2015,192:7.
• [8]DuráM A,Flores M,ToldráF.Purification and characterisation of a glutaminase from Debaryomyces spp[J].International Journal of Food Microbiology,2002,76(1-2):117.
• [9]Sabu A,Keerthi T R,Kumar S R,et al.L-Glutaminase production by marine Beauveriasp.under solid state fermentation[J].Process Biochemistry,2000,35(7):705-710.
• [10]Imada A,Igarasi S,Nakahama K,et al.Asparaginase and glutaminase activies of microorganisms[J].Journal of General Microbiology,1973,76(1):85-99.
• [11]Itoh T,Yasuto H,Matsuura S,et al.Production of L-theanine using glutaminase encapsulated in carbon-coated mesoporous silica with high pH stability[J].Biochemical Engineering Journal,2012,68(18):207-214.
• [12]Tomita K,Yano T,Kitagata T,et al.Formation ofγ-Glutamyl peptidesbyglutaminaseofAspergillus oryzae[J].Agricultural and Biological Chemistry,1989,53(7):1995-1996.
• [13]浦荷芳.硝基还原假单胞菌谷氨酰胺酶基因的克隆、表达及其在L-茶氨酸酶法合成中的应用[D].南京:南京师范大学,2012.
• [14]崔春,徐明,杨娟,等.离子选择电极法测定谷氨酰胺酶活力研究[J].中国调味品,2017,42(2):10-14.
• [15]Alexandra W Z,Christiane G D,Michael A.Functional characterization of a salt-and thermotolerant glutaminase from Lactobacillus rhamnosus[J].Enzyme and Microbial Technology,2003,32(7):862-867.
• [16]王雪燕,师文钊.蛋白质化学及其应用[M].北京:中国纺织出版社,2016:35-60.
• [17]Moriguchi M,Sakai K,Tateyama R,et al.Isolation and characterization of salt-tolerant glutaminases from marine Micrococcus luteus K-3[J].Journal of Fermentation&Bioengineering,1994,77(6):621-625.
• [18]康维民,刘绍军,常学东,等.一种新型谷氨酰胺酶在酱油酿造中的应用[J].中国调味品,2003(4):33-35.
• [19]Klein M,Kaltwasser H,Jahns T.Isolation of a novel,phosphateactivated glutaminase from Bacillus pasteurii[J].FEMS Microbiology Letters,2002,206(1):63-67.
• [20]Shigekezu Y,Aki K,Kazuaki Y,et al.Analysis of essential amino acid residues for catalytic activity of glutaminase from Micrococcus luteus K-3[J].Journal of Bioscience and Bioengineering,2006,102(4):362-364.
• [21]Yoshimune K,Shirakihara Y,Wakayama M,et al.Crystal structure of salt-tolerant glutaminase from Micrococcus luteus K-3in the presence and absence of its product L-glutamate and its activator Tris[J].FEBS Journal,2010,277(3):738-748.