人工设计二硫键增强谷氨酰胺转胺酶热稳定性

Enhancement of the Thermostability of Transglutaminase from Streptomyces hygroscopicus by Engineering a Disulfide Bond

DOI：10.3969/j.issn.1673-1689.2015.10.009

中文关键词: 谷氨酰胺转胺酶 二硫键 热稳定性

英文关键词: transglutaminase,disulfide bond,thermostability

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中文摘要:

对S. hygroscopicus来源的谷氨酰胺转胺酶（microbial transglutaminase，MTG） 进行分子改造，在MTG的N端区域引入二硫键，增强其热稳定性。通过序列比对与结构分析、运用Disulfide by Design软件，构建含有二硫键的突变体MTG（4-284）。在55 ℃下处理10 min，MTG（4-284）可以保持95%的酶活，而原始酶MTG仅剩15%的酶活。圆二色谱检测结果显示MTG和MTG（4-284）的T50分别为58和65 ℃。以上结果对MTG的分子水平的深入研究提供了基础，同时进一步满足其工业化应用的需求。

英文摘要:

In this study, transglutaminase (MTG) from S. hygroscopicus was molecularly modified to improve the thermostability. Mutant MTG (4-284) containing a disulfide bond in the N-terminal region was constructed through sequence aligment, structure analysis and the software Disulfide by Design. After treatment at 55 ℃ for 10 min, MTG (4-284) remained 95% activity whereas MTG only retained 15% activity. The T50 values of MTG and MTG (4-284) were 58 and 65 ℃, respectively. Results in this study could improve genetic reconstruction and promote the industry application of MTG.

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