食品科学

• 生物工程 • 上一篇    下一篇

β-淀粉酶的表达与酶学性质

张萧萧1,牛丹丹1,2,*,沈 微3,李 玉1,路福平1   

  1. 1.天津科技大学生物工程学院,天津 300457;2.福州大学生物科学与工程学院,福建 福州 350108;
    3.江南大学生物工程学院,江苏 无锡 214036
  • 出版日期:2016-02-15 发布日期:2016-02-26

Expression and Enzymatic Properties of β-Amylase

ZHANG Xiaoxiao1, NIU Dandan1,2,*, SHEN Wei3, LI Yu1, LU Fuping1   

  1. 1. College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China; 2. College of Biological Science and
    Engineering, Fuzhou University, Fuzhou 350108, China; 3. School of Biotechnology, Jiangnan University, Wuxi 214036, China
  • Online:2016-02-15 Published:2016-02-26

摘要:

本研究在构建重组表达质粒pPIC-BBA的基础上,构建并筛选获得了高分泌表达大麦β-淀粉酶的重组巴斯德毕赤酵母GS-BBA。重组酵母在摇瓶发酵条件下,能够分泌表达180 U/mL的β-淀粉酶酶活力。进一步对该重组酶的酶学性质进行了分析,重组β-淀粉酶的最适作用温度为55 ℃,最适作用pH值为5.0,在不高于55 ℃和在pH 3.0~10.0之间有良好的热稳定性和pH值稳定性。重组酶水解淀粉的主要产物为麦芽糖,在普鲁兰酶的协助下,其水解淀粉形成麦芽糖的最大转化率为78.28%。

关键词: &beta, -淀粉酶, 表达, 酶学性质, 麦芽糖

Abstract:

In this research, a recombinant Pichia pastoris strain GS-BBA overexpressing barley β-amylase was developed
using the recombinant expression plasmid pPIC-BBA. Strain GS-BBA could produce approximately 180 U/mL β-amylase in
shaking flask culture. The biochemical properties of the recombinant β-amylase were examined. The maximum activity was
achieved under the conditions of pH 5.0 and 55 ℃. Meanwhile, its excellent thermostability and pH stability was observed at
a temperature not exceeding 55 ℃ and in the pH range of 3.0–10.0. The recombinant enzyme could hydrolyze starch to form
maltose as the major product. With the aid of pullulanase, it hydrolyzed starch to form maltose with a conversion efficiency
up to 78.28%.

Key words: β-amylase, expression, enzymatic properties, maltose

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