48 Scopus citations

Abstract

For rapid analysis of microbial metabolisms,13C-fingerprinting employs a set of tracers to generate unique labeling patterns in key amino acids that can highlight active pathways. In contrast to rigorous 13C-metabolic flux analysis (13C-MFA), this method aims to provide metabolic insights without expensive flux measurements. Using13C-fingerprinting, we investigated the metabolic pathways in Rhodococcus opacus PD630, a promising biocatalyst for the conversion of lignocellulosic feedstocks into value-added chemicals. Specifically, seven metabolic insights were gathered as follows: (1) glucose metabolism mainly via the Entner-Doudoroff (ED) pathway; (2) lack of glucose catabolite repression during phenol co-utilization; (3) simultaneous operation of gluconeogenesis and the ED pathway for the co-metabolism of glucose and phenol; (4) an active glyoxylate shunt in acetate-fed culture; (5) high flux through anaplerotic pathways (e.g., malic enzyme and phosphoenolpyruvate carboxylase); (6) presence of alternative glycine synthesis pathway via glycine dehydrogenase; and (7) utilization of preferred exogenous amino acids (e.g., phenylalanine). Additionally, a13C-fingerprinting kit was developed for studying the central metabolism of non-model microbial species. This low-cost kit can be used to characterize microbial metabolisms and facilitate the design-build-test-learn cycle during the development of microbial cell factories.

Original languageEnglish
Pages (from-to)91-100
Number of pages10
JournalBiotechnology and Bioengineering
Volume113
Issue number1
DOIs
StatePublished - Jan 1 2016

Keywords

  • C-MFA
  • Catabolite repression
  • Entner-Doudoroff (ED) pathway
  • Gluconeogenesis
  • Phenol

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