Identification of the phd gene cluster responsible for phenylpropanoid utilization in Corynebacterium glutamicum

Appl Microbiol Biotechnol. 2016 Feb;100(4):1871-1881. doi: 10.1007/s00253-015-7165-1. Epub 2015 Nov 26.

Abstract

Phenylpropanoids as abundant, lignin-derived compounds represent sustainable feedstocks for biotechnological production processes. We found that the biotechnologically important soil bacterium Corynebacterium glutamicum is able to grow on phenylpropanoids such as p-coumaric acid, ferulic acid, caffeic acid, and 3-(4-hydroxyphenyl)propionic acid as sole carbon and energy sources. Global gene expression analyses identified a gene cluster (cg0340-cg0341 and cg0344-cg0347), which showed increased transcription levels in response to phenylpropanoids. The gene cg0340 (designated phdT) encodes for a putative transporter protein, whereas cg0341 and cg0344-cg0347 (phdA-E) encode enzymes involved in the β-oxidation of phenylpropanoids. The phd gene cluster is transcriptionally controlled by a MarR-type repressor encoded by cg0343 (phdR). Cultivation experiments conducted with C. glutamicum strains carrying single-gene deletions showed that loss of phdA, phdB, phdC, or phdE abolished growth of C. glutamicum with all phenylpropanoid substrates tested. The deletion of phdD (encoding for putative acyl-CoA dehydrogenase) additionally abolished growth with the α,β-saturated phenylpropanoid 3-(4-hydroxyphenyl)propionic acid. However, the observed growth defect of all constructed single-gene deletion strains could be abolished through plasmid-borne expression of the respective genes. These results and the intracellular accumulation of pathway intermediates determined via LC-ESI-MS/MS in single-gene deletion mutants showed that the phd gene cluster encodes for a CoA-dependent, β-oxidative deacetylation pathway, which is essential for the utilization of phenylpropanoids in C. glutamicum.

Keywords: Aromatics; Corynebacterium glutamicum; Degradation pathways; Lignin; Phenylpropanoids; β-oxidation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzene Derivatives / metabolism*
  • Carbon / metabolism
  • Cinnamates / metabolism*
  • Corynebacterium glutamicum / genetics*
  • Corynebacterium glutamicum / metabolism*
  • Energy Metabolism
  • Gene Deletion
  • Gene Expression Profiling
  • Gene Expression Regulation, Bacterial
  • Metabolic Networks and Pathways / genetics*
  • Multigene Family*

Substances

  • Benzene Derivatives
  • Cinnamates
  • Carbon