Purification and characterization of transglutaminase from a newly isolated Streptomyces hygroscopicus
Purification and characterization of transglutaminase from a newly isolated Streptomyces hygroscopicus
复制标题
DOI：
10.1016/j.foodchem.2007.04.020
复制
发表时间：
2007-01-01
期刊：
影响因子：
8.8
通讯作者：
Chen, Jian
中科院分区：
文献类型：
--
作者：
Cui, Li;Du, Guocheng;Chen, Jian
通过乙醇沉淀从培养液中纯化从土壤中分离的吸水链霉菌菌株的转谷氨酰胺酶（TGase，EC 2.3.2.13），然后在 CM-纤维素和 Sephadex G-75 柱上连续进行色谱分离，产率和纯化倍数分别为 21.1% 和 30%。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳估计该酶的分子量为38,000 Da。纯化的微生物转谷氨酰胺酶 (MTG) 在 37-45°C 和 pH 6.0-7.0 范围内表现出对 N-羧基苯甲酰基-L-谷氨酰胺酰-甘氨酸和羟胺形成异羟肟酸的最佳活性。该酶在50℃以上不稳定，在较低温度下在pH 5.0-8.0范围内稳定。 MTG 不受 Ca2+ 和乙二胺四乙酸的抑制，表明它不依赖于钙。纯化的 MTG 被 5,5'-二硫双（2-硝基苯甲酸）、Cu2+、Zn2+、Pb2+ 和 Hg2+ 强烈灭活，表明该酶在活性位点可能具有硫醇基团。 MTG 稳定性受乙醇浓度的强烈影响。在 25 摄氏度下，酶活性在较低浓度的乙醇下略有升高。(c) 2007 Elsevier Ltd. 保留所有权利。
Transglutaminase (TGase, EC 2.3.2.13) from a Streptomyces hygroscopicus strain isolated from soil was purified from culture broth by ethanol precipitation, followed by successive chromatographies on CM-cellulose and Sephadex G-75 columns with a yield and purification-fold of 21.1% and 30%, respectively. The enzyme's molecular weight was estimated as 38,000 Da by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified microbial transglutaminase (MTG) exhibited optimum activity at 37-45 degrees C and in a range of pH 6.0-7.0 for hydroxamate formation from N-carboxybenzoyl-L-glutaminyl-glycine and hydroxylamine. The enzyme was not stable above 50 degrees C and was stable within a pH range of 5.0-8.0 at lower temperature. The MTG was not inhibited by Ca2+ and ethylenediaminetetraacetic acid, suggesting it was calcium-independent. Purified MTG was strongly inactivated by 5,5'-dithiobis (2-nitrobenzoic acid), Cu2+, Zn2+, Pb2+, and Hg2+, suggesting that this enzyme could possess a thiol group at the active site. The MTG stability was strongly affected by ethanol concentration. The enzyme activity was slightly elevated at a lower concentration of ethanol at 25 degrees C. (c) 2007 Elsevier Ltd. All rights reserved.