作　　者： (何思雨）; (成文玉）; (金红星）;

机构地区： 河北工业大学化工学院,天津300130

出　　处： 《生物技术通报》 2017年第8期174-179,共6页

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 △fab D,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。 To increase rifamycin yield,the Amycolatopsis mediterranei mutant of the inactivated S-malonyltransferase gene（fab D）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 fab D-inactivated strain was screened by apramycin as amarker. Further,the rifamycin production between mutant strains was wmpared and the parental strain by fermentation. The results showedthat the homologous recombinant vector of fab D was successfully constructed,and the A. mediterranei △ fab D was obtained;the production ofrifamycin SV in the mutant strain was 168.08 mg/L,with 9.94% increase in contrast to the parent strain. The inactivation of S-malonyltransferasegene weakened the fatty acid synthesis of mutant strain,subsequently strengthened the synthesis of rifamycin.

关 键 词： 地中海拟无枝酸菌 利福霉素 丙二酰转移酶 融合 基因失活