龟裂链霉菌zwf2基因敲入及阻断对土霉素合成影响.doc

龟裂链霉菌zwf2基因敲入及阻断对土霉素合成影响龟裂链霉菌zwf2基因敲入及阻断对土霉素合成影响摘要,目的考察敲入及阻断zwf2基因对6-磷酸葡糖脱氢酶,G6PDH,活性、细胞生长、土霉素合成的影响。方法构建敲入转化子M4018-(zwf2)和阻断转化子M4018-Δzwf2后,以原始菌株为对照,比较敲入转化子菌株M4018-(zwf2)和阻断转化子菌株M4018-Δzwf2摇瓶发酵后G6PDH活性、细胞生长、土霉素合成等生物性质的变化。结果阻断转化子菌株M4018-Δzwf2的G6PDH活性是原始菌的50%左右,敲入转化子菌株M4018-(zwf2)的G6PDH活性则略高于原始菌,阻断转化子菌株M4018-Δzwf2土霉素生物合成水平比原始菌提高27%,比敲入转化子菌株M4018-(zwf2)高约40%,发酵结束时敲入转化子菌株M4018-(zwf2)的细胞干重高于其他2株菌,其它2株菌的细胞干重差异不大。结论zwf2阻断有利于土霉素的生物合成,zwf2增强则对细胞生长有利。关键词,6-磷酸葡糖脱氢酶,zwf2,龟裂链霉菌M4018,土霉素生物合成中图分类号,,A文章编号,11672-979X(2009)01-0007-04Effectsofzwf2GeneKnock-inandDisruptiononOxytetracyclineBiosynthesisinStreptomycesrimosusM4018LIAOYu-ling1,LIUZhi-yong2,TANGZhen-yu2,GUOMei-jin1,2*,ZHUANGYing-ping2,ZHANGSi-liang2(,JiangxiAgriculturalUniversity,Nanchang330045,China;,EastChinaUniversityofScience&Technology,Shanghai200237,China)Abstract,ObjectiveToinvesgatetheeffectsofzwf2geneknock-inanddisruptiononthespecificactivityofglucose-6-phosphatedehydrogenase(G6PDH),cellgrowthandoxytetracycline(OTC),thechangesofthespecificacitivityofG6PDH,cellgrowthandOTClevelinthemediaweremeasuredafterthemutantsM4018-Δzwf2andM4018-(zwf2)respectivelyobtainedthroughgenedisruptionandknock-