摘要
将编码黄孢原毛平革菌木质素过氧化物酶(lip)的cDNA克隆到酵母整合型质粒pMETA上,电转化Ade缺陷型甲醇毕赤酵母(Pichiamethanolica)PMAD16,通过MD平板及PCR方法筛选和鉴定重组子。重组子发酵液经SDSPAGE分析和木质素过氧化物酶活力测定等方法鉴定,表明带自身信号肽的黄孢原毛平革菌木质素过氧化物酶基因(lip)在甲醇毕赤酵母中得到表达。优化其发酵培养条件,以藜芦醇为底物进行酶活测定,其酶活可达932U/L。相应发酵指数为12.94U/h·L。比出发菌株提高了24.18%。
The gene of lignin peroxidase from phanerochaete chrysosporium (lip) was coloned and transformed into Ade defective mutant yeast PMAD16.Recombinant yeast strains were obtained by screening on MD plates and PCR technique.The expression products of lip gene were analyzed by SDS-PAGE and assay of lignin peroxidase. The results indicated that the lip gene from P.chrysosporium was successfully transformed and overexpressed in Pichia methanolica. Some factors such as media,buffer,pH,methanol concentration, media volume, inoculated volume were investigated to optimize the cultivation condition for expression of lip.Under the optimized conditions, the expression level of lip was up to 932U/L and the fermentation coefficient was 12.94U/h.L.
出处
《工业微生物》
CAS
CSCD
北大核心
2005年第1期10-15,共6页
Industrial Microbiology
