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当前位置: 首页 > 文章 > 吸水链霉菌谷氨酰胺转胺酶分离纯化方法改进及结构研究 生物技术通报 2011,0 (4) 199-203
Position: Home > Articles > Improvement of Purification Method and Study on Structure of Transglutaminase from Streptomyces hygroscopicus Biotechnology Bulletin 2011,0 (4) 199-203

吸水链霉菌谷氨酰胺转胺酶分离纯化方法改进及结构研究

作  者：
杨晓燕;周哲敏;堵国成;陈坚
单  位：
江南大学工业生物技术教育部重点实验室
关键词：
吸水链霉菌;谷氨酰胺转胺酶;分离纯化;结构
摘  要：
利用改进的新方法,对吸水链霉菌谷氨酰胺转氨酶(TGase)进行分离纯化。将发酵上清液经过硫酸铵分级盐析后,用Hitrap Q HP阴离子交换柱除掉干扰较大的色素,之后又经Superdex 75 10/300GL凝胶柱和Hitrap Q HP阴离子交换柱的分离,得到高纯度的TGase。纯化后TGase的比活力可达24.5 U/mg,回收率为39.9%。TGase的N-末端前6个氨基酸经测序为DAADER。该研究是对吸水链霉菌TGase的N-末端氨基酸序列的首次报道。将吸水链霉菌TGase的N-末端氨基酸序列与已报道的其它3种链霉菌来源的TGase的N-端氨基酸序列进行比对,序列相似性不高。预测和分析了吸水链霉菌TGase的高级结构,为进一步研究TGase结构和功能的关系,蛋白分子定向改造提供理论依据。
译  名：
Improvement of Purification Method and Study on Structure of Transglutaminase from Streptomyces hygroscopicus
作  者：
Yang Xiaoyan1 Zhou Zhemin1 Du Guocheng1 Chen Jian1,2(1The Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122; 2State Key Laboratory of Food Science and Technology,Jiangnan University,Wuxi 214122)
关键词：
Streptomyces hygroscopicus Transglutaminase Purification Structure
摘  要：
Transglutaminase(TGase)was purified from Streptomyces hygroscopicus fermentation liquid by an improved method.After ammonium sulfate precipitation,most of the pigment was removed from the enzyme solution using Hitrap Q HP anion exchange chromatography.The high purity TGase was obtained by further purification using Superdex 75 10/300GL and Hitrap Q HP columns,respectively.The Specific activity of purified enzyme was 24.5 U/mg,as well as the yield of 39.9%.N-terminal amino acid sequence of TGase was determined as DAADER.This is the first report of the N-terminal amino acid sequence of TGase from Streptomyces hygroscopicus.According to the amino acid sequence alignment,the N-terminal sequence of TGase from Streptomyces hygroscopicus exhibited low sequence similarity to those from other three Streptomyces species.Advanced structure of TGase from Streptomyces hygroscopicus was predicted and analysed,which provide theoretical evidence for further research on the relationship between structure and function and its directional modification.

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