摘要
多杀菌素是由刺糖多孢菌(Saccharopolyspora spinosa SIPI—A.2090)产生的重要农用抗生素,其生物合成途径已被阐明。NDP-葡萄糖合成酶(gtt)与葡萄糖脱氢酶(gdh)是多杀菌素生物合成途径中的限速酶。从SIPI-A.2090克隆gtt及gdh基因,并构建了表达这两个基因的整合型质粒,转入产多杀菌素刺糖多孢菌,发酵并验证其基因型。结果表明,阳性突变株SIPI—M.2092的多杀菌素发酵单位比出发菌株提高了173%,增加gtt和gdh基因拷贝数可以有效提高多杀菌素的发酵单位。
Spinosad was a secondary metabolite produced by the soil actinomycetes of Saccharopolyspora spinosa and its biosysthesis pathway of spinosyn was well revealed. Both NDP-glucose synthase （ gtt ） and glucose dehydratase （ gdh ） were rate-limiting enzymes. The gtt and gdh gene were isolated from S. spinosa SIPI-A. 2090 and then cloned into pOJ260 with a ermE＊ promoter upstream to construct a integration plasmid pYG1156. The plasmid was then transformed into S. sp/nosa SIPI-A. 2090. Based on homologous recombination and apramycin resistantance selection, a mutant SIPI-M. 2092 was obtained and its spinosad production was 173% higher than that of the original strain. The genome of the SIPI-M. 2092 was identificated correctly by PCR.
出处
《工业微生物》
CAS
CSCD
2010年第6期1-5,共5页
Industrial Microbiology
基金
"重大新药创制"科技重大专项项目(2009ZX09301-007]
