食品与发酵工业 >

2021 , Vol. 47 >Issue 17: 41 - 46

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026977

研究报告

基于液滴微流控的产α-淀粉酶地衣芽孢杆菌高通量筛选

  • 陈宇锟 ,
  • 黎青华 ,
  • 周景文 ,
  • 张国强 ,
  • 李江华
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  • 1(粮食发酵工艺与技术国家工程实验室(江南大学),江苏 无锡,214122)
    2(江南大学 生物工程学院,江苏 无锡,214122)
硕士研究生(张国强副研究员和李江华教授为共同通讯作者,E-mail:gqzhang@jiangnan.edu.cn;lijianghua@jiangnan.edu.cn)

收稿日期: 2021-02-03

  修回日期: 2021-03-08

  网络出版日期: 2021-09-27

基金资助

江苏省自然科学基金青年基金项目(BK20190607);江苏省自然科学基金前沿引领技术基础研究专项(BK20202002)

收起

High-throughput screening of α-amylase-producing Bacillus licheniformis based on droplet microfluidic system

  • CHEN Yukun ,
  • LI Qinghua ,
  • ZHOU Jingwen ,
  • ZHANG Guoqiang ,
  • LI Jianghua
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  • 1(National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi 214122,China)
    2(School of Biotechnology,Jiangnan University,Wuxi 214122,China)

Received date: 2021-02-03

  Revised date: 2021-03-08

  Online published: 2021-09-27

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摘要

α-淀粉酶广泛应用于食品、纺织等行业,市场潜力巨大。地衣芽孢杆菌 (Bacillus licheniformis) 是α-淀粉酶的主要生产菌株,但由于存在产酶能力低等问题,不能满足工业需求。为了获得高产α-淀粉酶的地衣芽孢杆菌生产菌株,将酶活性与荧光信号偶联,通过将高活性单细胞制备、液滴包埋以及筛选条件等参数优化,建立了基于液滴微流控技术的高通量筛选方法。基于此方法,对地衣芽孢杆菌BLH的突变体库进行高通量筛选,成功获得5株高产菌株,其发酵酶活力较出发菌株提高48%~66%。该研究建立的液滴微流控高通量筛选方法为高产淀粉酶地衣芽孢杆菌的选育提供选择,也为其他高性能蛋白表达分泌宿主的高效筛选提供了参考。

关键词： α-淀粉酶; 地衣芽孢杆菌; 荧光检测; 液滴微流控; 高通量筛选

本文引用格式

陈宇锟 , 黎青华 , 周景文 , 张国强 , 李江华 . 基于液滴微流控的产α-淀粉酶地衣芽孢杆菌高通量筛选[J]. 食品与发酵工业, 2021 , 47(17) : 41 -46 . DOI: 10.13995/j.cnki.11-1802/ts.026977

Abstract

high-throughput screening method for obtaining α-amylase high yield strain was developed and optimized. The screening process as following: single-cell preparation, droplet generation, and fluorescent coupled droplet microfluidic screening. Five high-yielding strains selected from the mutant library with α-amylase activity increased by 48%-66%. The droplet microfluidic high-throughput screening method provides an efficient method for the selection of high-yielded α-amylase B. licheniformis, and it is also conductive to the efficient screening for other hosts with high level of proteins expression and secretion.

Key words： α-amylase; Bacillus licheniformis; fluorescence detection; droplet microfluidic system; high-throughput screening

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