解脂耶氏酵母中羽扇豆醇合成途径的构建与调控

Construction and regulation of lupeol synthesis pathway in Yarrowia lipolytica

羽扇豆醇因其具有抗癌抗炎等生理活性而广泛应用于医药领域。本研究分别利用源自木榄和蓖麻的羽扇豆醇合酶(LUS)在解脂耶氏酵母(Yarrowia lipolytica)中构建生物合成羽扇豆醇途径(GLU-1、GLU-2),并由对该途径中关键限速酶3-羟基-3-甲基戊二酰辅酶a还原酶(tHMGR)和异戊烯基二磷酸异构酶(IDI)过表达的工程菌(GLU-1M、GLU-2M)调控MVA途径,实现了羽扇豆醇在解脂耶氏酵母中从无到有的生成,其产量为11.74 mg/L。为提高其产量而构建了过表达角鲨烯合成酶(SQS)及角鲨烯氧化酶(SQE)的工程菌(GLU-1MSS),羽扇豆醇的产量达到了24.62 mg/L,较前构建的工程菌(GLU-1M)产量提高了1倍。

Lupeol is widely used in the field of medicine,because of its anti-cancer and anti-inflammatory properties.In this study,lupeol synthase(LUS)derived from Bruguiera gymnorhiza and Ricinus communis was used to construct biosynthesis pathway of lupeol in Yarrowia lipolytica(GLU-1,GLU-2).Meanwhile,the MVA pathway was regulated by the key rate-limiting enzymes 3-hydroxy-3-methylglutaryl coenzyme reductase(tHMGR)and isopentenyl diphosphate isomerase(IDI)overexpressed in engineering bacteria(GLU-1M,GLU-2M).And,the production of lupeol in Y.lipolytica realized from nothing to 11.74 mg/L.In order to increase its yield,the engineering bacterium(GLU-1MSS)overexpressing squalene synthase(SQS)and squalene oxidase(SQE)was constructed.The lupeol yield could reach 24.62 mg/L,which was twice as high as that of the previously constructed engineering bacterium(GLU-1M).