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食品与发酵工业  2019, Vol. 45 Issue (2): 1-6    DOI: 10.13995/j.cnki.11-1802/ts.016703
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
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1, 王玉川1, 易萍2, 闫巧娟2, 江正强1*
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)
High level expression of α-amylase from Malbranchea cinnamomea in Pichia pastoris and preparation of maltose syrup
ZHAO Ning1, WANG Yuchuan1, YI Ping2, YAN Qiaojuan2, JIANG Zhengqiang1*
1(College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China);
2(College of Engineering, China Agricultural University, Beijing 100083, China)
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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。
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赵宁
王玉川
易萍
闫巧娟
江正强
关键词:  樟绒枝霉  α-淀粉酶  毕赤酵母  高密度发酵  麦芽糖浆    
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.
Key words:  Malbranchea cinnamomea    α-amylase    Pichia pastoris    high-level expression    maltose syrup
收稿日期:  2018-01-04                出版日期:  2019-01-25      发布日期:  2019-02-21      期的出版日期:  2019-01-25
基金资助: 国家重点研发计划2018YFD0400404
作者简介:  硕士研究生(江正强教授为通讯作者,E-mail:zhqjiang@cau.edu.cn)。
引用本文:    
赵宁,王玉川,易萍,等. 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.
ZHAO Ning,WANG Yuchuan,YI Ping,et al. High level expression of α-amylase from Malbranchea cinnamomea in Pichia pastoris and preparation of maltose syrup[J]. Food and Fermentation Industries, 2019, 45(2): 1-6.
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http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.016703  或          http://sf1970.cnif.cn/CN/Y2019/V45/I2/1
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