Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 113, Issue 1, Pages 91-100Publisher
WILEY
DOI: 10.1002/bit.25702
Keywords
C-13-MFA; catabolite repression; Entner-Doudoroff (ED) pathway; gluconeogenesis; phenol
Categories
Funding
- NSF [DBI 1356669]
- DOE [DE-SC0012705]
- Div Of Biological Infrastructure
- Direct For Biological Sciences [1356669] Funding Source: National Science Foundation
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For rapid analysis of microbial metabolisms,C-13-fingerprinting employs a set of tracers to generate unique labeling patterns in key amino acids that can highlight active pathways. In contrast to rigorous C-13-metabolic flux analysis (C-13-MFA), this method aims to provide metabolic insights without expensive flux measurements. Using(13)C-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, a(13)C-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. Biotechnol. Bioeng. 2016;113: 91-100. (c) 2015 Wiley Periodicals, Inc.
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