单宁酶基因在黑曲霉中的同源表达

Tannase gene homologous expressing in Aspergillus niger

利用PCR方法从黑曲霉基因组中扩增单宁酶基因的编码区，构建其黑曲霉表达载体pSZH-tan。通过农杆菌介导法将TAN基因导入黑曲霉中，经筛选获得将TNA基因整合到糖化酶基因的同源重组转化子。对转化子表达产物进行SDS-PAGE分析和酶活检测，结果表明，重组蛋白分子质量约为76 ku，重组菌株单宁酶表达量为322~581μg·mL^-1，最高发酵酶活为41.12 U·mL^-1。

The tannase coding sequence was amplified from Aspergil us niger by the method of PCR, and then the expression vector PSZH-tan was constructed. The TAN gene transfer by Agrobacterium-medi-ated Aspergil us niger, the TNA was screened glucoamylase gene into the gene homologous recombinants. Expression product was analyzed by SDS-PAGE and the enzyme activity was detected, the result showed that the recombinant protein molecular weight was about 76 ku, expression of recombinant strains tannase was 322-581μg·mL^-1, the maximum fermentation activity was 41.12 U·mL^-1.