Primers were designed according to the whole genome sequencing annotation.The key de-gumming enzyme genes (pectinase gene,xylanase gene and mannanase gene)were cloned from the strain Dickeya sp.DCE -01,and recombined into the expression vector pET -28a and transformed into E.coli BL21.The results showed that pel E was 1185 bp,encoded 395 amino acids,and the molecular weight of the protein was 44 kDa;Xyn was 1251 bp,encoded 416 amino acids,the molecular weight of the protein was 45.74 kDa;Man was 1137 bp,encoded 379 amino acids,the molecular weight of the pro-tein was 41.85 kDa.The enzymatic activity of pectinase, xylanase and mannanase reached 471.97 U /mL,64.32 U /mL and 16882.86 U /mL,correspondingly.This study would provide a scientif-ic basis for exploring the genetic resources from the strain DCE -01 and developing efficient industrial enzyme preparation.%根据全基因组测序注释结果设计特异引物,从DCE-01菌株克隆关键脱胶酶基因:果胶酶基因(pel E)、木聚糖酶基因(xyn)及甘露聚糖酶基因(man),重组到表达载体p ET-28a中,导入E.coli BL21中进行诱导表达.结果表明:pel E核酸序列长1185 bp,编码395个氨基酸,蛋白分子量为44 kDa;xyn核酸序列长1251 bp,编码416个氨基酸,蛋白分子量45.74 kDa;man核酸序列长1137 bp,编码379个氨基酸,蛋白分子量41.85 kDa.果胶酶、木聚糖酶、甘露聚糖酶的酶活力分别为471.97、64.32、16882.86 U/mL.该研究为进一步发掘DCE-01菌株基因资源及开发高效工业酶制剂提供了科学依据.
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