单宁酶基因在黑曲霉ST31中的克隆与表达

利用PCR扩增得到米曲霉(Aspergillusoryzae)单宁酶(tannase)基因的编码序列,经DNA测序证实单宁酶基因已成功克隆,然后将其连接到黑曲霉的表达载体ANED2-SP2上构建单宁酶基因表达载体。将构建好的单宁酶基因表达载体通过原生质转化法导入黑曲霉菌株ST31中进行表达研究。结果表明重组菌株的单宁酶活力最高为104.02U/ml发酵液,比原始出发菌株米曲霉提高2~3倍。研究构建了黑曲霉的高效转化体系,提高了黑曲霉表达系统的应用水平,为其它新酶的研究提供有价值的参考。

Cloning and Expressing of Tannase Gene from Aspergillus oryzae in Aspergillus niger ST31

It is the first time to research about expression of heterologous tannase gene using Aspergillus niger expression system. The gene encoding Tannase was cloned from A. oryzae by PCR. After tannase was sequenced, expression vector ANEP2-SP2-tan was constructed and was transformed into A. niger ST31 by protoplast transformation. Then, assaying of recombinant tannase activity were done. So, a novel recombinant Aspergillus strain containing tannase gene was obtained. Maximal tannase activity was 104.02 U/ml. It is as 2 to 3 times as the original strain. Based on the study, high efficient transformation system of A. niger is constructed and applied latitude of A. niger expression system was broadened. It is also useful for further working in research of new enzymes.