Engineering the lva operon and Optimization of Culture Conditions for Enhanced Production of 4-Hydroxyvalerate from Levulinic Acid in Pseudomonas putida KT2440

Abstract

Monomeric 4-hydroxyvalerate is a versatile chemical used to produce various commodities and fine chemicals. In the present study, the lvaAB gene was deleted from the lva operon in Pseudomonas putida KT2440 and tesB, obtained from Escherichia coli, was overexpressed under the control of the lva operon system, which is induced by the substrate levulinic acid and the product 4-hydroxyvalerate to produce 4-hydroxyvalerate from levulinic acid. The lvaAB-deleted strain showed almost complete conversion of levulinic acid to 4-hydroxyvalerate, compared with 24% conversion in the wild-type strain. In addition, under optimized culture conditions, the final engineered strain produced a maximum of 50 g/L 4-hydroxyvalerate with 97% conversion from levulinic acid. The system presented here could be applied to produce high titers of 4-hydroxyvalerate in a cost-effective manner at a large scale from renewable cellulosic biomass.