4-Hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF), also called homo furaneol or 2 (or 5)- ethyl-4-hydroxy-5 (or 2)-methyl-3(2H)-furanone, is an important industrial aromatic compound that exhibits the unique characteristic of keto–enol tautomerism. HEMF is a safe edible flavoring agent approved for use by the American Flavor and Extract Manufacturers Association of the United States (FEMA) and has been developed as a raw flavoring material. Owing to its unique fragrance, HEMF is widely used as a food flavoring agent and has high demand. Enone oxidoreductase is a vital enzyme involved in HEMF production. In this study, an enone oxidoreductase from Naumovozyma dairenensis CBS 421 (NDEO) was used for HEMF production for the first time. The mutant NDEOT183W, K290W was obtained through semirational protein engineering, which increased HEMF yield by 75.2%. Finally, the engineered strain BM4 produced the highest HEMF yield, 194.42 mg L-1 in 132h, this yield was 7.2-fold higher than that of BY4741. Very few studies have reported on microbial production of HEMF. We believe that our proposed strategy will be helpful in accelerating the industrial production of HEMF and realizing the commercialization of more high-value natural products in the future.

Screening and remodeling of enone oxidoreductase for high production of 2(or 5)-ethyl-5(or 2)-methyl-4-hydroxy-3(2H)-furanone in Saccharomyces cerevisiae

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