EngineeringEscherichia colifor the production of butyl octanoate from endogenous octanoyl-CoA

Abstract

Medium chain esters produced from fruits and flowering plants have a number of commercial applications including use as flavour and fragrance ingredients, biofuels, and in pharmaceutical formulations. These esters are typically made via the activity of an alcohol acyl transferase (AAT) enzyme which catalyses the condensation of an alcohol and an acyl-CoA. Developing a microbial platform for medium chain ester production using AAT activity presents several obstacles, including the low product specificity of these enzymes for the desired ester and/or low endogenous substrate availability. In this study, we engineeredEscherichia colifor the production of butyl octanoate from endogenously produced octanoyl-CoA. This was achieved through rational protein engineering of an AAT enzyme fromActinidia chinensisfor improved octanoyl-CoA substrate specificity and metabolic engineering ofE. colifatty acid metabolism for increased endogenous octanoyl-CoA availability. This resulted in accumulation of 3.3 + 0.1 mg/L butyl octanoate as the sole product fromE. coliafter 48 h. This study represents a preliminary examination of the feasibility of developingE. coliplatforms for the synthesis single medium chain esters from endogenous fatty acids.