黑曲霉单宁酶基因Tan2克隆与表达

Cloning and Expression of Tannase Gene Tan2 from Aspergillus niger

单宁酶(Tannase,EC 3.1.1.20)能水解单宁中的酯键和羧酚酸键,产生没食子酸以及对应醇,在食品、饮料、饲料、制药、医药、化妆品等各类工业中应用广泛,也在普洱茶发酵中具有重要作用。从普洱茶发酵中分离的黑曲霉菌株PU001中克隆得到单宁酶基因Tan2,并连接到表达载体pCold-Ⅰ构建BL21-pColdⅠ原核冷诱导表达系统,转化至感受态大肠杆菌BL21中,SDS-PAGE与质谱鉴定均表明单宁酶Tan 2表达成功,旨在为进一步研究该酶在普洱茶发酵中的作用机制以及其他领域的应用奠定基础。

Tannase(EC 3.1.1.20)can hydrolyze the ester bond and carboxyphenolic acid bond in tannins to produce gallic acid and corresponding alcohols.It is widely used in food,beverage,feed,pharmacy,medicine,cosmetics and other industries,and also plays an important role in the fermentation of Pu-erh tea.In this work,tannase gene Tan 2 was cloned from Aspergillus niger PU001,which was previously isolated from fermentation in Pu-erh tea.It was ligated with expression vector pCold-Ⅰ to construct the prokaryotic cold inducible expression system of BL21-pCold Ⅰ,which was transformed into Escherichia coli BL21.SDS-PAGE and mass spectrometry identification showed that tannase Tan 2 was expressed successfully.This result provides a basis for further survey on the role of tannase in the fermentation of Pu-erh tea and application in other fields.