食品与发酵工业 >

2022 , Vol. 48 >Issue 3: 44 - 49

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.028423

研究报告

常压室温等离子诱变选育L-色氨酸高产菌株

  • 周旭波 ,
  • 陈瑜琦 ,
  • 丁鼎 ,
  • 刘帅 ,
  • 王健
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  • (江苏澳创生物科技有限公司,江苏 无锡,214000)
博士研究生,高级工程师(王健教授为通信作者,E-mail:wj@acbiotech.ltd)

收稿日期: 2021-06-21

  修回日期: 2021-07-28

  网络出版日期: 2022-03-04

基金资助

2019年江苏省“双创计划”项目(苏财行[2019]60号)

收起

Breeding of L-tryptophan high-yield strain by ARTP mutagenesis

  • ZHOU Xubo ,
  • CHEN Yuqi ,
  • DING Ding ,
  • LIU Shuai ,
  • WANG Jian
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  • (Jiangsu AC Biotech Co.Ltd.,Wuxi 214000,China)

Received date: 2021-06-21

  Revised date: 2021-07-28

  Online published: 2022-03-04

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摘要

为了提升发酵法产L-色氨酸的生产效率,使用常压室温等离子(atmospheric and room temperature plasma,ARTP)诱变育种技术,并结合结构类似物抗性定向筛选的方法,选育高产色氨酸菌株。将出发菌Escherichia coli AC-1042经过ARTP诱变处理后,在抗性培养基平板上筛选具有5-甲基色氨酸、对氟苯丙氨酸抗性的突变菌株,选取产酸高、遗传稳定的菌株重复进行ARTP诱变处理和抗性筛选,不断提高菌株对结构类似物的抗性水平。经过多次ARTP诱变处理和抗性筛选,获得1株色氨酸高产菌株ACTRP104,经过30 L发酵罐培养44 h后L-色氨酸质量浓度可以达到61.65 g/L,葡萄糖转化率达到20.64%,比出发菌分别提高了20.69%和17.81%。结果表明,ARTP诱变和结构类似物抗性筛选相结合,可以有效地获得色氨酸高产突变菌株,大大提高色氨酸的发酵生产技术水平,获得的色氨酸高产菌株ACTRP104具有较好的工业化应用前景。

关键词： 常压室温等离子体(ARTP); L-色氨酸; 诱变育种; 抗性筛选; 高产突变菌株

本文引用格式

周旭波 , 陈瑜琦 , 丁鼎 , 刘帅 , 王健 . 常压室温等离子诱变选育L-色氨酸高产菌株[J]. 食品与发酵工业, 2022 , 48(3) : 44 -49 . DOI: 10.13995/j.cnki.11-1802/ts.028423

Abstract

To improve the efficiency of L-tryptophan production, atmospheric and room temperature plasma (ARTP) mutagenesis combined with analogue resistance screening was used to obtain high-yield L-tryptophan production strain. Escherichia coli AC-1042 was mutated by ARTP mutagenesis, and the L-tryptophan genetic stable high-yield strain was selected by 5-methyltryptophan (5-MT) and p-fluorophenylalanine (PFP) resistance. After multiple ARTP mutagenesis processing and improving the analogue resistance screening, a genetic stable high-yield tryptophan strain ACTRP104 was obtained. The tryptophan mutant ACTRP104 produced 61.65 g/L L-tryptophan at a yield of 20.64% of glucose in the 30 L fermentation tank at 44 h, which was increased by 20.69% and 17.81% compared with the original strain, respectively. These results suggested that it was effective to improve the production capacity of tryptophan strains by combining of ARTP mutagenesis with analogue resistance screening. The tryptophan high-yield mutant ACTRP104 has a great potential for industrial application.

Key words： atmospheric and room temperature plasma (ARTP); L-tryptophan; mutation breeding; resistance screening; high-yield mutant

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