文献类型：期刊文献

中文题名：Cyclobacterium qasimii脱乙酰酶的异源表达及酶学性质

英文题名：Heterologous Expression and Enzymatic Properties of N-Acetylglucosamine Deacetylase from Cyclobacterium qasimii

作者：马佳菲[1];秦臻[2];范立强[1];赵黎明[1];蒋丽华[1]

机构：[1]华东理工大学生物工程学院/发酵工业分离提取技术研发中心/生物反应器工程国家重点实验室,上海200237;[2]上海大学生命科学学院,上海201900

年份：2020

卷号：38

期号：6

起止页码：39

中文期刊名：食品科学技术学报

外文期刊名：Journal of Food Science and Technology

收录：CSTPCD;;EI(收录号：20212010368178);Scopus;北大核心:【北大核心2017】；CSCD:【CSCD_E2019_2020】；

基金：国家重点研发计划课题(2019YFD0901805);上海市曙光计划项目(15SG28);高等学校学科创新引智计划项目(B18022)。

语种：中文

中文关键词：Cyclobacterium qasimii;氨基葡萄糖;N-乙酰氨基葡萄糖;脱乙酰酶;异源表达;酶学性质

外文关键词：Cyclobacterium qasimii;glucosamine;N-acetylglucosamine;deacetylase;heterologous expression;enzymatic properties

摘要：氨基葡萄糖因具有丰富的生物活性而被广泛应用,但目前其工业化生产均需利用浓酸在高温条件下进行脱乙酰反应;因此,需要寻找一种环境友好的生物催化法来取代酸水解法脱乙酰基。筛选并得到了来源于卡西氏菌Cyclobacterium qasimii的脱乙酰酶(CqCBDA),并将其成功地在大肠杆菌表达系统中进行了异源表达,且其具有较高水平的可溶性蛋白表达量。纯化后重组的CqCBDA具有N-乙酰氨基葡萄糖脱乙酰酶活性,比酶活力为11.5 U/mg,最适温度为40℃,最适pH值为7.6,在低于40℃的条件下展现出较高的热稳定性。CqCBDA在酶解法生产氨基葡萄糖过程中具有较大的应用潜力,研究结果可为酶法绿色制备氨基葡萄糖提供理论参考。
Glucosamine is widely used because of its diverse biological activity.However,its industrial production requires the hydrolysis reaction with concentrated acid at high temperature.Therefore,an environmentally friendly biocatalytic method is needed to replace the deacetylation of glucosamine by acid hydrolysis.In this study,the deacetylase(CqCBDA)from Cyclobacterium qasimii was screened and was successfully expressed heterologously in the Escherichia coli,which had a high level of soluble protein expression.The purified recombinant CqCBDA had the N-acetylglucosamine deacetylation activity and the specific enzyme activity was 11.5 U/mg.The optimal temperature and pH value of CqCBDA was 40℃and 7.6,while CqCBDA showed high thermal stability below 40℃.CqCBDA has great potential to be applied to the production of glucosamine by enzymatic hydrolysis.This study provides theoretical reference for green manufacturing of glucosamine by the enzymatic approach.