生物学杂志 ›› 2023, Vol. 40 ›› Issue (1): 40-.doi: 10.3969/j.issn.2095-1736.2023.01.040

• 研究报告 • 上一篇    下一篇

rRNA甲基转移酶LmrB基因突变对林可霉素生物合成的影响

许玉荣1, 许婉莲2, 徐晶晶3, 赵 明1, 高 亮2, 吴 杭2   

  1. 1. 合肥师范学院 化学与化学工程学院, 合肥 230601; 2. 安徽大学 生命科学学院 
    物质科学与信息技术研究院, 合肥 230601; 3. 中国科学技术大学附属第一医院, 合肥 23000
  • 出版日期:2023-02-18 发布日期:2023-02-21
  • 通讯作者: 吴杭,博士,教授,博士生导师,研究方向为微生物代谢工程,E-mail: wuhang@ahu.edu.cn
  • 作者简介:许玉荣,博士,讲师,主要从事抗生素代谢工程研究,E-mail: xuyurong89@163.com
  • 基金资助:
    安徽省自然科学基金项目(2008085QC99); 合肥师范学院高层次人才科研启动基金项目(2020rcjj36)和省级科研平台专项项目(2020PT20); 安徽省高校自然科学基金重点项目(KJ2019A0720); 国家级大学生创新创业训练计划资助项目(202114098020S)

Impact of gene mutation of the rRNA methyltransferase LmrB on the lincomycin biosynthesis in Streptomyces lincolnensis #br#

XU Yurong1, XU Wanlian2, XU Jingjing3, ZHAO Ming1, GAO Liang2, WU Hang2   

  1. 1.Department of Chemical and Chemical Engineering, Hefei Normal University, Hefei 230601,
    China; 2.School of Life Sciences, Anhui University, Hefei 230601,China; 3.The First Affiliated Hospital of
    Anhui Provincial Hospital, Hefei 230001, China
  • Online:2023-02-18 Published:2023-02-21

摘要: 为探究林可链霉菌野生菌NRRL2936和高产菌LC-G中lmrB基因的抗性以及对林可霉素产量的影响,将来自两种不同菌株的lmrB(2936-B和LC-G-B)连接到表达载体pET28a和pIB139上,分别转入异源宿主大肠杆菌BL21与变铅青链霉菌TK24中,获得相应的lmrB异源表达菌株,对其进行林可霉素抗性实验。通过同源重组方法,在NRRL2936和LC-G中分别缺失lmrB,并在LC-G中进行lmrB的过表达,对所获得的重组菌株进行林可霉素产量检测。结果显示:尽管LC-G-B基因片段长度短于2936-B,但并未使LC-G-B丧失功能,且在大肠杆菌和变铅青链霉菌中,2936-B对林可霉素的抗性均明显高于LC-G-B;同时,lmrB缺失突变株△lmrB的林可霉素A产量较NRRL2936降低约40%。在LC-G中缺失lmrB基因,林可霉素A产量降低约30%,而将NRRL2936的lmrB引入到LC-G中可显著提高林可霉素的产量。

关键词: 林可链霉菌, 林可霉素, rRNA甲基转移酶, 抗性

Abstract: In order to explore the resistance function and the effect on lincomycin production of lmrB gene in Streptomyces lincolnensis NRRL2936 and LC-G,the lmrB gene from two different strains was ligated to different expression vectors pET28a and pIB139. The resistance test was carried out in E.coli BL21 and Streptomyces lividans TK24, respectively. lmrB gene in NRRL2936 and LC-G was respectively deleted by homologous recombination, and was over expressed in LC-G.The lincomycin production of S. lincolnensis and its derivatives was detected by HPLC. The results showed that although the length of LC-G-B gene fragment was shorter than that of 2936-B, LC-G-B did not lose its function, and the resistance of 2936-B to lincomycin was significantly higher than that of LC-G-B in E. coli and S.lividans. Meanwhile, the lincomycin yield of lmrB deletion mutant △lmrB was about 40% lower than that of NRRL2936. With the deletion of lmrB gene in LC-G, the yield of lincomycin was decreased by about 30%.The introduction of lmrB from NRRL 2936 into high-yield strain LC-G can significantly improve the yield of lincomycin.

Key words: Streptomyces lincolnensis, lincomycin, rRNA methyltransferase, resistance

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