生物技术通报 ›› 2017, Vol. 33 ›› Issue (8): 174-179.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0200

• 研究报告 • 上一篇    下一篇

S-丙二酰转移酶基因失活对地中海拟无枝酸菌产利福霉素的影响

何思雨, 成文玉, 金红星   

  1. 河北工业大学化工学院,天津 300130
  • 收稿日期:2017-03-17 出版日期:2017-08-01 发布日期:2017-08-01
  • 作者简介:何思雨,女,硕士,研究方向:微生物遗传育种学;E-mail:hesiyu2017@126.com

Effects of S-malonyltransferase Gene Inactivation on Rifamycin Production by Amycolatopsis mediterranei

HE Si-yu, CHENG Wen-yu, JIN Hong-xing   

  1. School of Chemical Engineering and Technology,Hebei University of Technology,Tianjin 300130
  • Received:2017-03-17 Published:2017-08-01 Online:2017-08-01

摘要: 为提高利福霉素的产量,构建了S-丙二酰转移酶基因失活的地中海拟无枝酸菌。利用融合PCR构建S-丙二酰转移酶基因的同源重组载体,通过电击转化导入到地中海拟无枝酸菌中,使之发生同源重组,以安普霉素为标记,筛选了S-丙二酰转移酶基因失活菌株,并对比了突变菌株与原始菌株的利福霉素SV产量。成功构建了S-丙二酰转移酶基因的同源重组载体,获得了地中海拟无枝酸菌突变株A. mediterranei △fabD,失活菌株的利福霉素SV产量为168.08 mg/L,比原始菌提高了9.94%。S-丙二酰转移酶基因的失活,弱化了突变菌株脂肪酸的合成,强化了利福霉素的合成。

关键词: 地中海拟无枝酸菌, 利福霉素, S-丙二酰转移酶, 融合PCR, 基因失活

Abstract: To increase rifamycin yield,the Amycolatopsis mediterranei mutant of the inactivated S-malonyltransferase gene(fabD)was constructed. S-malonyltransferase gene homologous recombinant vector was constructed using fusion PCR,and then transformed into A. mediterranei by electroporation,making the homologous recombination occurr. Then fabD-inactivated strain was screened by apramycin as a marker. Further,the rifamycin production between mutant strains was wmpared and the parental strain by fermentation. The results showed that the homologous recombinant vector of fabD was successfully constructed,and the A. mediterranei △fabD was obtained;the production of rifamycin SV in the mutant strain was 168.08 mg/L,with 9.94% increase in contrast to the parent strain. The inactivation of S-malonyltransferase gene weakened the fatty acid synthesis of mutant strain,subsequently strengthened the synthesis of rifamycin.

Key words: Amycolatopsis mediterranei, rifamycin, S-malonyltransferase, fusion PCR, gene inactivation