Bioproduction of 4-vinylphenol from corn cob alkaline hydrolyzate in two-phase extractive fermentation using free or immobilized recombinant E. coli expressing pad gene

In situ extractive fermentation was used to produce 4-vinyl derivatives from hydroxycinnamic acids extracted from corn cobs by recombinant Escherichia coli cells expressing Lactobacillus plantarum phenolic acid descarboxylase (PAD) gene. This microorganism mainly produced 4-vinylphenol (4VP) from p-coumaric acid (p-CA). In a first study, we observed that concentrations of 4VP higher than 1 g/L had a negative impact on descarboxylation of p-CA to 4VP by recombinant E.coli cells. Because of this, and in order to improve the downstream process, a two-phase aqueous-organic solvent system was developed. The results of the extractive fermentation indicated that it was possible to use hydrolyzates as aqueous phase to bioproduce 4VP, and recover simultaneously the product in the organic phase containing hexane. The detoxification of pre-treated corn cob alkaline hydrolyzate improved 4VP production up to 1003.5 mg/L after 24 h fermentation (QP = 41.813 mg/L·h). Additionally, preliminary experiments using cells immobilized in calcium alginate showed to be a good system for the biotransform of p-CA to 4VP in extractive fermentation, although the process hindered partially the recovery of 4VP in the organic phase.We are grateful to the Spanish Ministry of Science and Innovation for the financial support of this work (CTQ2011-28967 and AGL2011-22745 projects), which has partial financial support from the FEDER funds of the European Union. J.M.S. is grateful for Postdoctoral fellowship (SFRH / BPD / 84440 / 2012) of Fundacao para a Ciencia e Tecnologia - FCT