摘要
目的对弗氏链霉菌(Streptomyces fradiae)泰乐菌素还原酶(tylosin reductase,SFR)进行改造以抑制其对泰乐菌素(tylosin)的还原作用,提高泰乐菌素发酵产量和产品质量。方法在前期对泰乐菌素还原酶编码基因第642位碱基进行定点突变形成终止密码子的基础上,对该基因642位后碱基进行敲除;对基因敲除菌株的生长以及发酵产物中泰乐菌素(A组分)和雷诺菌素(relomycin,D组分)含量进行分析。结果获得了泰乐菌素还原酶基因部分序列敲除的弗氏链霉菌菌株M1-d1;该菌株培养过程中生长状态及稳定期生物量与原始菌株基本一致,菌落形态正常;泰乐菌素发酵产物中敲除菌株A组分相对含量较原始菌株相比增加了10.08%,而D组分相对含量较原始菌株减少68.75%。结论弗氏链霉菌泰乐菌素还原酶基因部分序列敲除不影响菌体正常生长,但能够抑制泰乐菌素还原酶对A组分的还原作用,提高泰乐菌素发酵产物中A组分含量,显著降低D组分含量。
Objective For the purpose of inhibiting reduction activity of tylosin reductase and improving yield and product quality in tylosin fermentation,the tylosin reductase of Streptomyces fradiae was modified in this study.Methods On the basis of a single base mutation of the 642th base in tylosin reductase gene that lead to forming a termination codon,the bases after the 642th position in the gene were knocked out.Then,the growth and fermentation of the gene knockout strains were performed.The contents of tylosin(A)and renomycin(D)in the product were analyzed.Results The Streptomyces fradiae strain M1-d1 with sequence knockout after the 642th base of tylosin reductase gene was obtained.The growth state and biomass in the stable phase of the strain were basically the same as the original strain,and the colony morphology was normal.Compared with the original strain,the relative content of component A in the knockout strain increased by 10.08%,while the relative content of component D in the fermentation product decreased by 68.75%.Conclusion The partial sequence knockout of tylosin reductase gene did not affect the growth of Streptomyces fradiae,but inhibited the tylosin reductase activity on component A.As a result,content of component A in tylosin fermentation product was increased while the component D was decreased significantly.
作者
蒋玉婷
朱慧
贾晨波
岳思君
苏建宇
Jiang Yu-ting;Zhu Hui;Jia Chen-bo;Yue Si-jun;Su Jian-yu(Key Laboratory of Education for Protection and Utilization of Special Biological Resources in Western China,College of Life Sciences,Ningxia University,Yinchuan 750021)
出处
《中国抗生素杂志》
CAS
CSCD
北大核心
2023年第5期518-523,共6页
Chinese Journal of Antibiotics
基金
宁夏回族自治区重点研发计划项目(No.2021BEG02003)。
作者简介
蒋玉婷,女,生于1996年,在读硕士研究生,主要研究方向为微生物资源开发与利用,E-mail:375927216@qq.com;通讯作者:苏建宇,E-mail:su_jy@nxu.edu.cn。
