CAJ | 학술논문

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。
이용PCR방법종흑곡매기인조중확증단저매기인적편마구，구건기흑곡매표체재체pSZH-tan。통과농간균개도법장TAN기인도입흑곡매중，경사선획득장TNA기인정합도당화매기인적동원중조전화자。대전화자표체산물진행SDS-PAGE분석화매활검측，결과표명，중조단백분자질량약위76 ku，중조균주단저매표체량위322~581μg·mL-1，최고발효매활위41.12 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.