烯还原酶不对称还原(R)-香芹酮的研究  

Study on asymmetric reduction of ( R )-carvone by ene-reductase

作　　者：鄢洪德 YAN Hongde(College of Pharmaceutical Engineering and Biotechnology,Zhejiang Pharmaceutical University,Ningbo 315199,China)

机构地区：[1]浙江药科职业大学制药工程与生物技术学院,浙江宁波315199

出　　处：《发酵科技通讯》2024年第2期74-78,共5页Bulletin of Fermentation Science and Technology

摘　　要：(2R,5R)-二氢香芹酮是一种重要的手性中间体,可用于合成多种活性化合物,常采用不对称还原(R)-香芹酮的方法合成。相较于化学方法,烯还原酶催化还原(R)-香芹酮不需要危险的氢气、贵金属催化剂和高温等苛刻的条件,且来源于Corynebacterium casei的烯还原酶(CcER)能有效地还原(R)-香芹酮产生(2R,5R)-二氢香芹酮。考察了pH、反应温度、辅酶NAD~+浓度和辅底物葡萄糖浓度对反应的影响,研究表明最佳反应条件为pH 8、反应温度30℃、NAD^(+)浓度0.6 mmol/L、葡萄糖浓度30 mmol/L;反应6 h后,(R)-香芹酮的转化率可达96%,产物(2R,5R)-二氢香芹酮的光学纯度可达95%;烯还原酶催化能力强,具有生产(2R,5R)-二氢香芹酮的潜力。(2R,5R)-Dihydrocarvone stands as a pivotal chiral intermediate,facilitating the synthesis of numerous active compounds.Its production often involves the asymmetric reduction of(R)-carvone.Notably,the utilization of enereductases for this reduction process eliminates the need for stringent conditions such as hazardous hydrogen,precious metals,and elevated temperatures.Specifically,the ene-reductase sourced from Corynebacterium casei(Cc ER)demonstrates a notable ability to efficiently reduce(R)-carvone,yielding(2R,5R)-dihydrocarvone.In-depth studies focused on optimizing reaction parameters,including pH,reaction temperature,coenzyme NAD+concentration,and cosubstrate glucose concentration.The optimal reaction conditions were identified at pH 8 and reaction temperature 30℃,with a NAD^(+)concentration of 0.6 mmol/L and a glucose concentration of 30 mmol/L.Under these conditions,the conversion of(R)-carvone reached an impressive 96%,with a diastereomeric excess of 95%toward the desired product,(2R,5R)-dihydrocarvone,achieved within a reaction time of 6 hours.The remarkable catalytic performance of Cc ER underscores its potential for efficiently producing(2R,5R)-dihydrocarvone.

关 键 词：烯还原酶 (2R 5R)-二氢香芹酮 氧化还原酶 立体选择性 

分 类 号：Q814.9[生物学—生物工程]