环糊精葡萄糖基转移酶的分子改造及其在合成α-熊果苷中的应用  

Molecular modification of cyclodextrin glucosyltransferase and its application in the synthesis ofα-arbutin

作　　者：刘嘉琦 谭明[2] 董钧 刘以银 刘珊娜 吴洽庆[2] LIU Jiaqi;TAN Ming;DONG Jun;LIU Yiyin;LIU Shanna;WU Qiaqing(College of Food Science and Bioengineering,Tianjin Agricultural University,Tianjin 300392,China;Tianjin Institute of Industrial Biology,Chinese Academy of Sciences,Tianjin 300308,China)

机构地区：[1]天津农学院食品科学与生物工程学院,天津300392 [2]中国科学院天津工业生物技术研究所,天津300308

出　　处：《生物工程学报》2024年第6期1845-1855,共11页Chinese Journal of Biotechnology

基　　金：国家重点研发计划(2021YFC2102000)。

摘　　要：熊果苷在化妆品和医药领域具有重要应用价值,但从植物组织中提取产率低,极大地限制了其应用。本研究利用戈特沙尔克厌氧分支杆菌(Anaerobranca gottschalkii)来源的环糊精葡萄糖基转移酶(cyclodextrin glucosyltransferase,CGTase),以麦芽糊精为供体,对苯二酚为受体,催化合成α-熊果苷。通过定点饱和突变和定点突变筛选到突变体Ag CGTase-F235G-N166H,活力是野生型的3.48倍。通过优化反应pH、温度、对苯二酚添加量,转化率在最优反应条件下达到63%。综上所述,本研究成功构建了一株能高效合成α-熊果苷并具有较高苯二酚转化率的菌株,对于降低α-熊果苷工业化的生产成本,提高产物的转化率方面具有重要的应用潜力。α-arbutin has important applications in cosmetics and medicine.However,the extraction yield from plant tissues is relatively low,which restricts its application value.In this study,we investigated the synthesis ofα-arbutin using maltodextrin as the donor and hydroquinone as the acceptor,using a cyclodextrin glucosyltransferase(CGTase)from Anaerobranca gottschalkii.We performed site-saturated and site-directed mutagenesis on AgCGTase.The activity of the variant AgCGTase-F235G-N166H was 3.48 times higher than that of the wild type.Moreover,we achieved a conversion rate of 63%by optimizing the reaction pH,temperature,and hydroquinone addition amount.Overall,this study successfully constructed a strain with improved conversion rate for the synthetic production ofα-arbutin and hydroquinone.These findings have significant implications for reducing the industrial production cost ofα-arbutin and enhancing the conversion rate of the product.

关 键 词：环糊精葡萄糖基转移酶 α-熊果苷 定点突变 麦芽糊精 对苯二酚 

分 类 号：Q814[生物学—生物工程] TQ426.97[化学工程]