Currently, the biotechnological preparation of fragrances using natural materials attracted growing attention. Enzymatic synthesis of vanillin from isoeugenol by recombinant isoeugenol monooxygenase from Pseudomonas nitroreducens Jin1 was systematically investigated herein. With series of work on the construction of recombinant E. coli over-expressing isoeugenol monooxygenase, optimization of the culture conditions for enzyme production and reaction process for converting isoeugenol into vanillin, an increase of 22-fold in the enzyme activity (2050 U/L) was obtained, and the conversion was significantly increased at high substrate concentration with the aid of magnetic chitosan membrane for product isolation in situ. Under optimal conditions, the product concentration and space-time yield reached 252 mM and 115 g/L/d, respectively, and vanillin was obtained in 82.3% yield and > 99% purity in the gram preparative scale. The developed bioprocess showed application potential for efficient preparation of vanillin from inexpensive natural resources.

当前，使用天然材料的香料的生物技术制备引起了越来越多的关注。本文系统地研究了通过硝化假单胞菌Jin1的重组异丁香酚单加氧酶从异丁香酚的酶促合成香兰素。关于重组大肠杆菌构建的一系列工作过量表达异丁香酚单加氧酶，优化酶生产的培养条件和将异丁香酚转化为香兰素的反应过程，酶活性（2050 U / L）增加了22倍，并且在高温下转化率显着增加借助磁性壳聚糖膜对底物进行浓缩以原位分离产物。在最佳条件下，产物浓度和时空产率分别达到252 mM和115 g / L / d，以克制备规模获得香兰素的产率为82.3％，纯度> 99％。发达的生物工艺显示出从廉价的自然资源有效制备香兰素的应用潜力。