研究报告

樟绒枝霉α-淀粉酶在毕赤酵母中的高效表达及在麦芽糖浆制备中的作用

  • 赵宁 ,
  • 王玉川 ,
  • 易萍 ,
  • 闫巧娟 ,
  • 江正强
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  • 1(中国农业大学 食品科学与营养工程学院,北京,100083);
    2(中国农业大学 工学院,北京,100083)
硕士研究生(江正强教授为通讯作者,E-mail:zhqjiang@cau.edu.cn)。

收稿日期: 2018-01-04

  网络出版日期: 2019-02-21

基金资助

国家重点研发计划2018YFD0400404

High level expression of α-amylase from Malbranchea cinnamomea in Pichia pastoris and preparation of maltose syrup

  • ZHAO Ning ,
  • WANG Yuchuan ,
  • YI Ping ,
  • YAN Qiaojuan ,
  • JIANG Zhengqiang
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  • 1(College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China);
    2(College of Engineering, China Agricultural University, Beijing 100083, China)

Received date: 2018-01-04

  Online published: 2019-02-21

摘要

从嗜热真菌樟绒枝霉(Malbranchea cinnamomea)中克隆α-淀粉酶基因McAmyA,并在毕赤酵母GS115中高效表达,经高密度发酵至168 h时,胞外酶活力达到13 440.6 U/mL。重组α-淀粉酶McAmyA粗酶液经QSFF强阴离子交换层析纯化得到电泳级纯酶,比酶活力为1 230.2 U/mg。酶学性质研究表明,重组α-淀粉酶McAmyA的最适pH和最适温度分别是6.5和65 ℃。以淀粉液化液为底物,在温度60 ℃,加酶量120 U/g,水解24 h的条件下,重组α-淀粉酶McAmyA水解液化液,制备得到麦芽糖含量为50.0%(质量分数)的麦芽糖浆。该真菌α-淀粉酶在毕赤酵母中表达水平高,具有很大的应用潜力。

本文引用格式

赵宁 , 王玉川 , 易萍 , 闫巧娟 , 江正强 . 樟绒枝霉α-淀粉酶在毕赤酵母中的高效表达及在麦芽糖浆制备中的作用[J]. 食品与发酵工业, 2019 , 45(2) : 1 -6 . DOI: 10.13995/j.cnki.11-1802/ts.016703

Abstract

The α-amylase gene (McAmyA) from Malbranchea cinnamomea was successfully cloned and heterologously expressed in Pichia pastoris GS115. The recombinant α-amylase (McAmyA) was secreted at high level of 13440.6 U/mL by high cell density fermentation after 168 h. McAmyA was purified by QSFF ion-exchange chromatography with specific activity of 1230.2 U/mg. The optimal pH and temperature of McAmyA were determined to be 6.5 and 65 ℃, respectively. The maltose syrup with maltose content of 50% was prepared at 60 ℃, 120 U/g and 24 h. The fungus α-amylase was highly expressed in Pichia pastoris and it has great potential for application.

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