Production of Esters in Escherichia coli Using Citrate Synthase Variants

Author:

Shipmon Jacoby C.1,Rathinasabapathi Pasupathi12,Broich Michael L.1,Hemansi 13,Eiteman Mark A.1ORCID

Affiliation:

1. School of Chemical, Materials and Biomedical Engineering, University of Georgia, Athens, GA 30602, USA

2. Department of Genetic Engineering, SRM Institute of Science and Technology, Chengalpattu District, Kattankulathur 603202, Tamil Nadu, India

3. Department of Microbiology, Central University of Haryana, Mahendergarh 123029, Haryana, India

Abstract

Acetate esters comprise a wide range of products including fragrances and industrial solvents. Biosynthesis of esters offers a promising alternative to chemical synthesis because such routes use renewable carbohydrate resources and minimize the generation of waste. One biochemical method for ester formation relies on the ATF1 gene from Saccharomyces cerevisiae, which encodes alcohol-O-acyltransferase (AAT) which converts acetyl-CoA and an exogenously supplied alcohol into the ester. In this study, the formation of several acetate esters via AAT was examined in Escherichia coli chromosomally expressing citrate synthase variants, which create a metabolic bottleneck at acetyl-CoA. In shake flask cultures, variant strains generated more acetate esters than the strains expressing the wild-type citrate synthase. In a controlled bioreactor, E. coli GltA[A267T] generated 3.9 g propyl acetate in 13 h, corresponding to a yield of 0.155 g propyl acetate/g glucose, which is 18% greater than that obtained by the wild-type GltA control. These results demonstrate the ability of citrate synthase variants to redistribute carbon from central metabolism into acetyl-CoA-derived biochemicals.

Funder

United States National Science Foundation

Publisher

MDPI AG

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