食品与发酵工业 >

2021 , Vol. 47 >Issue 10: 159 - 163

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.025998

生产与科研应用

吡咯喹啉醌产生菌的筛选和菌种鉴定

  • 朱道洋 ,
  • 魏春雨 ,
  • 余晓斌
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  • (江南大学 生物工程学院,工业生物技术教育部重点实验室,江苏 无锡,214122)
硕士研究生(余晓斌教授为通讯作者,E-mail:xbyu@jiangnan.edu.cn)

收稿日期: 2020-10-25

  修回日期: 2020-12-15

  网络出版日期: 2021-06-17

基金资助

国家轻工技术与工程一流学科自主课题(LITE2018-11)

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Screening and identification of pyrroloquinoline quinone producing strains

  • ZHU Daoyang ,
  • WEI Chunyu ,
  • YU Xiaobin
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  • (School of Biotechnology, The Key Laboratory of Industrial Biotechnology, Ministry of Education,Jiangnan University, Wuxi 214122, China)

Received date: 2020-10-25

  Revised date: 2020-12-15

  Online published: 2021-06-17

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摘要

为了筛选产生吡咯喹啉醌(pyrroloquinoline quinone,PQQ)的菌株并提高PQQ发酵产量,该研究建立了一种快速的高效液相色谱法检测PQQ,并采用了以甲醇为唯一碳源的选择性培养基,利用光谱法对样品进行初筛,高效液相色谱法对初筛得到的结果再进行复筛。该研究还对筛选得到的菌株发酵产PQQ的培养基进行了优化。研究结果显示,从样品中筛选得到56株PQQ产生菌,摇瓶产量最高的1株为20.6 mg/L,通过16S rDNA 测序分析,鉴定该菌株为Methylopila sp.。对培养基成分中的碳源甲醇进行了优化研究,当甲醇体积分数为2.5%时,PQQ产量最高,为26.3 mg/L,相比于初始甲醇1.0%提高了28%。该研究建立的高效液相色谱法为PQQ高产菌株的筛选提供一种快速检测方法,筛选到的菌株具有较高的甲醇耐受力和PQQ生产潜力,为之后的PQQ发酵研究提供参考。

关键词： 吡咯喹啉醌; Methylopila sp.; 选择性培养基; 光谱法; 高效液相色谱法

本文引用格式

朱道洋 , 魏春雨 , 余晓斌 . 吡咯喹啉醌产生菌的筛选和菌种鉴定[J]. 食品与发酵工业, 2021 , 47(10) : 159 -163 . DOI: 10.13995/j.cnki.11-1802/ts.025998

Abstract

This study established rapid high performance liquid chromatography method for the detection of pyrroloquinoline quinone (PQQ). In order to screen the strains that produce PQQ, and increase the fermentation yield of PQQ, a selective medium with methanol was adopted as the sole carbon source. Spectroscopy was used for preliminary screening of the samples, HPLC was used for re-screening, and the fermentation medium of the selected strains to produce PQQ was optimized. The results showed that 56 PQQ-producing strains were screened from the samples, and the highest yield was 20.6 mg/L. Through 16s rDNA sequencing analysis, the strain was identified as Methylopila sp.. The methanol content in the fermentation medium was also optimized. When the methanol volume concentration was 2.5%, the PQQ yield reached 26.3 mg/L, which was 28% higher than that in the initial medium with 1.0% methanol. The HPLC method established in this study provided a rapid detection method for the screening of PQQ high-yielding strains. The selected strains had high methanol tolerance and PQQ production potential, which could provide a reference for further PQQ fermentation research.

Key words： pyrroloquinoline quinone(PQQ); Methylopila sp.; selective medium; spectroscopy; HPLC

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