茶树花青素还原酶是催化非酯型儿茶素EC和EGC合成的关键酶。采用RT-PCR技术,获得了茶树花青素还原酶基因的开放阅读框,它编码含337个氨基酸的蛋白质,推测分子量为37 kD,等电点为6.54;成功地将该基因重组到表达载体pET32a(+)上,并在大肠杆菌rosetta中进行原核表达;优化了原核表达中诱导时间、诱导温度、IPTG浓度、氨苄青霉素浓度,纯化出目的蛋白。HPLC检测表明,目的蛋白具有ANR酶活性。 Tea tree anthocyanin reductase is a key enzyme in the synthesis of non-ester catechins EC and EGC. The open reading frame of the anthocyanin reductase gene of tea plant was obtained by RT-PCR. It encodes a protein of 337 amino acids with a predicted molecular weight of 37 kD and an isoelectric point of 6.54. The gene was successfully cloned into the expression vector pET32a (+) and prokaryotic expressed in E. coli rosetta. The induction time, induction temperature, IPTG concentration and ampicillin concentration in prokaryotic expression were optimized and the target protein was purified. HPLC test showed that the target protein has ANR enzyme activity.