Dihydrocarvone, a natural terpene compound, has widespread applications in the fields of medicine, food, cosmetics, and chemical industries.It can be produced through the reduction of carvone catalyzed by ene reductase.However, the coenzyme NAD(P)H required for the reaction is expensive, serving as a restrictive factor for large-scale production.In this study, the glucose dehydrogenase (GDH) from Bacillus subtilis 9902 was cloned and expressed.Based on the previously obtained ene reductase KlebER1A303N from Klebsiella sp.O852, the co-expression system of KlebER1A303N and GDH in Escherichia coli was constructed using co-expression and fusion expression strategies.The differences in the expression of target proteins and the catalytic synthesis of dihydrocarvone among different strategies were compared and analyzed.The results showed that both co-expression and fusion expression could successfully express the desired target proteins and catalyze the conversion of carvone to dihydrocarvone.Nevertheless, the effects of fusion expression were superior to that of co-expression, with a conversion rate reaching 50% after 3 h of reaction.Additionally, the order of genes in fusion expression exerted a certain influence on the catalytic synthesis of dihydrocarvone.This study offers a foundation for establishing an efficient and low-cost industrial production method for dihydrocarvone.
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