生物学杂志

生物学杂志 ›› 2022, Vol. 39 ›› Issue (4): 33-.doi: 10.3969/j.issn.2095-1736.2022.04.033

• 研究报告 • 上一篇    下一篇

重离子诱变选育高产核黄素枯草芽孢杆菌

  

  1. HU Jianhua, ZHANG Zhaokun, LI Yongli, LAN Hui,LIU Zhanying
  • 出版日期:2022-08-18 发布日期:2022-08-15
  • 通讯作者: 刘占英，博士，教授，从事生物化工的研究，E-mail: liuzy1979@yeah.net
  • 作者简介:胡建华，博士，副教授，从事微生物代谢调控研究, E-mail: jhhu79@imut.edu.cn
  • 基金资助:
    内蒙古自治区科技成果转化项目（CGZH2018133）; 兰州重离子加速器国家实验室课题项目

Screening of high-yield riboflavin Bacillus subtilis strain by heavy-ion beam irradiation

  • Online:2022-08-18 Published:2022-08-15

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摘要： 研究以产核黄素的枯草芽孢杆菌C0为出发菌株，经12C6+重离子辐射诱变后，以8-氮鸟嘌呤作为拮抗物进行高产菌株的筛选，初步获得60个诱变菌株，进一步筛选得到4株高产菌株，最后经摇瓶发酵，发现4株诱变菌株中B13的核黄素产量最高为（401.8±9.2）mg/L，比原始菌株提高18.5%。经二次重离子诱变选育后，获得一株高产菌株BII7，核黄素产量达到（818.8±17.3）mg/L，比一次诱变菌株B13提高77.7%，表明重离子辐射对产核黄素的枯草芽孢杆菌具有良好的诱变效果。对诱变菌株BII7进行培养温度、pH值及转速等条件的优化，最适培养条件为37 ℃、pH 7.7、220 r/min。在5 L双联发酵罐中扩大培养高产菌株BII7，37 ℃、pH 7.7及溶氧浓度40%时，发酵时间缩短12 h，核黄素产量达1385.3 mg/L，比摇瓶培养提高69.2%，该诱变菌株具有较好的放大生产能力。6次传代培养实验表明该诱变高产菌株具有稳定的遗传特性。

关键词: 重离子, 诱变选育, 核黄素, 枯草芽孢杆菌

Abstract: The parent riboflavin-producing strain Bacillus subtilis C0 was treated with heavy-ion beam irradiation, then it was screened the mutagenic strains with 8-azaguanine. In further study, 4 high-yield strains were initially obtained from 60 mutant strains. After shake flask fermentation, the highest yield of mutant strain B13 reached( 401.8 ± 9.2) mg/L, which was 18.5% higher than that of the original strain. The new mutant strain BII7 was screened after second time of heavy-ion beam irradiation, and the riboflavin yield achieved (818.8±17.3) mg/L which was 77.7% higher than that of the mutation strain B13.The heavy-ion beam irradiation had good mutagenic effect. The optimal cultivation conditions of strain BII7 were: initial pH 7.7, culture temperature 37 ℃, and rotation speed 220 r/min. The riboflavin yield of mutant strain BII7 was 1 385.3 mg/L in 5 L fermenter (37 ℃, pH 7.7 and dissolved oxygen concentration 40%), which was 69.2% higher than that in shake flask. The fermentation time was shortened by 12 h. The results showed that the strain BII7 had good fermentation ability and stable genetic characteristics.

Key words: heavy-ion beam irradiation； mutation breeding, riboflavin, Bacillus subtilis

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