Abstract
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.
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Funding
This work was financially supported by Open Project Funding of the State Key Laboratory of Biocatalysis and Enzyme Engineering (No. SKLBEE2018008), Shanghai Collaborative Innovation Center of Fragrance Flavor and Cosmetics (No. 1021ZK170004002), and Shanghai Institute of Science and Technology (No. 10120K209007-A06).
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Yi Xu and Baodi Ma conceived and designed the study. Qian Wang and Xinyi Lu performed the experiments and analyzed the data. Qian Wang prepared the draft manuscript. Yi Xu, Baodi Ma, Yucai He, and Xiaomei Wu reviewed and edited the manuscript. All authors read and approved the manuscript.
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Wang, Q., Wu, X., Lu, X. et al. Efficient Biosynthesis of Vanillin from Isoeugenol by Recombinant Isoeugenol Monooxygenase from Pseudomonas nitroreducens Jin1. Appl Biochem Biotechnol 193, 1116–1128 (2021). https://doi.org/10.1007/s12010-020-03478-5
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DOI: https://doi.org/10.1007/s12010-020-03478-5