食品与发酵工业 >

2022 , Vol. 48 >Issue 16: 56 - 64

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.030228

研究报告

吡咯喹啉醌高产菌株选育及发酵优化

  • 张静 ,
  • 刘孟粟 ,
  • 秦志杰 ,
  • 曾伟主 ,
  • 周景文
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  • 1(江南大学, 粮食发酵工艺与技术国家工程实验室,江苏 无锡, 214122)
    2(江南大学, 未来食品科学中心,江苏 无锡, 214122)
硕士研究生(周景文教授为通信作者,E-mail:zhoujw1982@jiangnan.edu.cn)

收稿日期: 2021-11-26

  修回日期: 2021-12-28

  网络出版日期: 2022-09-16

基金资助

国家重点研发计划(2019YFA090490)

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Breeding and fermentation optimization of high titer pyrrorole-quinoline quinone strain

  • ZHANG Jing ,
  • LIU Mengsu ,
  • QIN Zhijie ,
  • ZENG Weizhu ,
  • ZHOU Jingwen
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  • 1(National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China)
    2(Science Center for Future Foods, Jiangnan University, Wuxi 214122, China

Received date: 2021-11-26

  Revised date: 2021-12-28

  Online published: 2022-09-16

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摘要

吡咯喹啉醌(pyrroloquinoline quinone,PQQ)是一种红褐色、芳香、三环邻醌的化合物,作为辅因子参与电子传递。在食品、医药和化妆品领域具有重要的应用价值。近几年,微生物发酵法合成PQQ受到了越来越多的关注。该研究以一株具有PQQ生产能力的脱氮生丝微菌(CGMCC 1.12893)为出发菌株,其在摇瓶水平上初始PQQ产量为31.4 mg/L。通过采用常压室温等离子体(atmospheric and room temperature plasma, ARTP)诱变和高通量筛选相结合的方法,筛选得到一株PQQ的高产突变菌株,编号为34,其PQQ产量提高至48.4 mg/L,相比原始菌株提高了54.1%。针对该最优突变菌株,在摇瓶水平上对碳源、氮源、金属离子和维生素等成分进行了优化,PQQ产量提高至105.2 mg/L。最后,在5 L发酵罐上对突变菌株进行验证培养,PQQ产量进一步提高至349.8 mg/L。该研究通过随机诱变筛选得到一株高产PQQ突变株,并通过培养基优化及发酵罐验证培养进一步提高了PQQ产量,研究结果为后续建立相应的发酵过程控制策略奠定了基础。

关键词： 脱氮生丝微菌; 吡咯喹啉醌; ARTP诱变; 高通量筛选; 发酵优化

本文引用格式

张静 , 刘孟粟 , 秦志杰 , 曾伟主 , 周景文 . 吡咯喹啉醌高产菌株选育及发酵优化[J]. 食品与发酵工业, 2022 , 48(16) : 56 -64 . DOI: 10.13995/j.cnki.11-1802/ts.030228

Abstract

Pyrroloquinoline quinone (PQQ) is a reddish-brown aromatic tricyclic ortho-quinone compound that functions as an electron transport cofactor. PQQ has high application value in food, medicine and cosmetics industries. In recent years, the production of PQQ by microbial fermentation methods has gained considerable interest. In this study, Hyphomicrobium denitrificans (CGMCC 1.12893) was selected as the starting strain, which could produce 31.4 mg/L PQQ at the shake flask scale. Through the combination of ARTP mutation and high-throughput screening method, a high-yielding mutant strain of PQQ was screened and numbered as 34, which accumulated 48.4 mg/L of PQQ, a 54.1% increase over the starting strain. For this strain, the components in the shake flask fermentation medium, including carbon source, nitrogen source, metal ions and vitamin were optimized, and the yield of PQQ increased to 105.2 mg/L. Finally, the production of PQQ was further increased to 349.8 mg/L in a 5 L bioreactor. In conclusion, the PQQ high-producing mutant strain was obtained through random mutagenesis screening, and production of PQQ was further improved through medium optimization in shake flask and fermentation in a 5 L bioreactor. The results laid a groundwork for the subsequent development of fermentation process control strategies.

Key words： Hyphomicrobium denitrificans; pyrrorolequinoline quinone; atmospheric and room temperature plasma (ARTP) mutagenesis; high-throughput screening; fermentation optimization

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