School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, PR China.
Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University, Tianjin 300350, PR China.
J Agric Food Chem. 2022 Aug 17;70(32):9888-9897. doi: 10.1021/acs.jafc.2c03306. Epub 2022 Aug 4.
Owing to its unique fragrance, 4-hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (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 CBS 421 (NDEO) was used for HEMF production for the first time. The mutant NDEO was obtained through semirational protein engineering, which increased the HEMF yield by 75.2%. Finally, the engineered strain BM4 produced the highest HEMF yield, 194.42 mg L in 132 h. Our study revealed that HEMF production can be improved in and that this is an efficient method to improve the activity of enone oxidoreductase, which is important for the industrial synthesis of furanone.
由于其独特的香气，4-羟基-2（或 5）-乙基-5（或 2）-甲基-3（2H）-呋喃酮（HEMF）被广泛用作食品调味剂，需求量很大。烯酮氧化还原酶是参与 HEMF 生产的一种重要酶。在这项研究中，首次使用来自 CBS 421（NDEO）的烯酮氧化还原酶来生产 HEMF。通过半理性蛋白质工程获得了突变体 NDEO，其将 HEMF 的产量提高了 75.2%。最后，工程菌株 BM4 在 132 小时内产生了最高的 HEMF 产量，为 194.42mg/L。我们的研究表明，可以提高 HEMF 的产量，这是一种有效提高烯酮氧化还原酶活性的方法，对呋喃酮的工业合成具有重要意义。
