Activation of the glutamic acid-dependent acid resistance system in Escherichia coli BL21(DE3) leads to increase of the fatty acid biotransformation activity

Ji Min Woo, Ji Won Kim, Ji Won Song, Lars M. Blank, Jin Byung Park

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29 Scopus citations

Abstract

The biosynthesis of carboxylic acids including fatty acids from biomass is central in envisaged biorefinery concepts. The productivities are often, however, low due to product toxicity that hamper whole-cell biocatalyst performance. Here, we have investigated factors that influence the tolerance of Escherichia coli to medium chain carboxylic acid (i.e., n-heptanoic acid)-induced stress. The metabolic and genomic responses of E. coli BL21(DE3) and MG1655 grown in the presence of n-heptanoic acid indicated that the GadA/B-based glutamic acid-dependent acid resistance (GDAR) system might be critical for cellular tolerance. The GDAR system, which is responsible for scavenging intracellular protons by catalyzing decarboxylation of glutamic acid, was inactive in E. coli BL21(DE3). Activation of the GDAR system in this strain by overexpressing the rcsB and dsrA genes, of which the gene products are involved in the activation of GadE and RpoS, respectively, resulted in acid tolerance not only to HCl but also to n-heptanoic acid. Furthermore, activation of the GDAR system allowed the recombinant E. coli BL21(DE3) expressing the alcohol dehydrogenase of Micrococcus luteus and the Baeyer-Villiger monooxygenase of Pseudomonas putida to reach 60% greater product concentration in the biotransformation of ricinoleic acid (i.e., 12-hydroxyoctadec-9-enoic acid (1)) into n-heptanoic acid (5) and 11-hydroxyundec-9-enoic acid (4). This study may contribute to engineering E. coli-based biocatalysts for the production of carboxylic acids from renewable biomass.

Original languageEnglish
Article numbere0163265
JournalPLoS ONE
Volume11
Issue number9
DOIs
StatePublished - 1 Sep 2016

Bibliographical note

Funding Information:
This study was supported by a Marine Biomaterials Research Center grant from the Marine Biotechnology Program funded by Ministry of Oceans and Fisheries, Republic of Korea No. D11013214H480000100 Dr. Jin-Byung Park Ministry of Education, Republic of Korea. Dr. Jin-Byung Park. This work was supported by the BK21 plus program through the National Research Foundation (NRF) funded by the Ministry of Education of Korea.

Publisher Copyright:
� 2016 Woo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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