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Title: Energetics of end product excretion in anaerboic bacteria and the metabolism of fatty acids by Syntrophomonas wolfei

Abstract

Anaerobic syntrophic bacteria degrade fatty acids which are important intermediates in anaerobic degradation and methanogenesis. These bacteria grow very slowly and require the presence of a hydrogen/formate-using organism to degrade fatty acids. Thus, these bacteria serve as models to study the biochemical aspects of mutualism and the energetics of slow growing organisms. We developed methods to physically separate cells of the anaerobic, fatty acid degrader, Syntrophomonas wolfei, from cells of the hydrogen user by Percoll gradient centrifugation and to selectively lyse S. wolfei cells using lysozyme. These methods allowed the study of the physiology of S. wolfei without significant contamination by cellular components of the hydrogen user. We also obtained pure cultures of S. wolfei by adapting the organism to grow on crotonate. Fatty acids were degraded by the {beta}-oxidation pathway using a coenzyme A (CoA) transferase activity to activate the fatty acid and substrate-level phosphorylation reactions to synthesize adenosine-5{prime}-triphosphate (ATP). The substrate specificity of the CoA transferase activity in the pure culture of S. wolfei differed from that found in the coculture suggesting that the ability to use crotonate resulted from an alteration of this enzyme. S. wolfei grown alone degraded crotonate in a manner similar to that ofmore » other crotonate-fermenting anaerobes, but the molar growth yields of S. wolfei were 2 to 3 times higher than those organisms. This suggests that the reduction of crotonyl-CoA to butyryl-CoA is energy yielding. S. wolfei contained a c-type cytochrome which may be involved in this reaction. S. wolfei synthesized large amounts of the storage polymer, poly-{beta}-hydroxybutyrate (PHB). Radioisotopic incorporation experiments showed that PHB was synthesized directly from the {beta}-oxidation intermediate rather than from the condensation of two acetyl-CoA molecules.« less

Authors:
Publication Date:
Research Org.:
Oklahoma Univ., Norman, OK (USA)
Sponsoring Org.:
DOE/ER
OSTI Identifier:
7013135
Report Number(s):
DOE/ER/13053-8
ON: DE90007549
DOE Contract Number:  
AS05-83ER13053
Resource Type:
Technical Report
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; METHANOGENIC BACTERIA; BIOLOGICAL PATHWAYS; TRANSFERASES; BIOCHEMICAL REACTION KINETICS; CARBON 14 COMPOUNDS; MONOCARBOXYLIC ACIDS; PROGRESS REPORT; TRACER TECHNIQUES; BACTERIA; CARBOXYLIC ACIDS; DOCUMENT TYPES; ENZYMES; ISOTOPE APPLICATIONS; KINETICS; LABELLED COMPOUNDS; MICROORGANISMS; ORGANIC ACIDS; ORGANIC COMPOUNDS; REACTION KINETICS; 090900* - Biomass Fuels- Processing- (1990-); 099000 - Biomass Fuels- Products & By-Products- (1990-); 550201 - Biochemistry- Tracer Techniques

Citation Formats

McInerney, M J. Energetics of end product excretion in anaerboic bacteria and the metabolism of fatty acids by Syntrophomonas wolfei. United States: N. p., 1989. Web.
McInerney, M J. Energetics of end product excretion in anaerboic bacteria and the metabolism of fatty acids by Syntrophomonas wolfei. United States.
McInerney, M J. Sun . "Energetics of end product excretion in anaerboic bacteria and the metabolism of fatty acids by Syntrophomonas wolfei". United States.
@article{osti_7013135,
title = {Energetics of end product excretion in anaerboic bacteria and the metabolism of fatty acids by Syntrophomonas wolfei},
author = {McInerney, M J},
abstractNote = {Anaerobic syntrophic bacteria degrade fatty acids which are important intermediates in anaerobic degradation and methanogenesis. These bacteria grow very slowly and require the presence of a hydrogen/formate-using organism to degrade fatty acids. Thus, these bacteria serve as models to study the biochemical aspects of mutualism and the energetics of slow growing organisms. We developed methods to physically separate cells of the anaerobic, fatty acid degrader, Syntrophomonas wolfei, from cells of the hydrogen user by Percoll gradient centrifugation and to selectively lyse S. wolfei cells using lysozyme. These methods allowed the study of the physiology of S. wolfei without significant contamination by cellular components of the hydrogen user. We also obtained pure cultures of S. wolfei by adapting the organism to grow on crotonate. Fatty acids were degraded by the {beta}-oxidation pathway using a coenzyme A (CoA) transferase activity to activate the fatty acid and substrate-level phosphorylation reactions to synthesize adenosine-5{prime}-triphosphate (ATP). The substrate specificity of the CoA transferase activity in the pure culture of S. wolfei differed from that found in the coculture suggesting that the ability to use crotonate resulted from an alteration of this enzyme. S. wolfei grown alone degraded crotonate in a manner similar to that of other crotonate-fermenting anaerobes, but the molar growth yields of S. wolfei were 2 to 3 times higher than those organisms. This suggests that the reduction of crotonyl-CoA to butyryl-CoA is energy yielding. S. wolfei contained a c-type cytochrome which may be involved in this reaction. S. wolfei synthesized large amounts of the storage polymer, poly-{beta}-hydroxybutyrate (PHB). Radioisotopic incorporation experiments showed that PHB was synthesized directly from the {beta}-oxidation intermediate rather than from the condensation of two acetyl-CoA molecules.},
doi = {},
url = {https://www.osti.gov/biblio/7013135}, journal = {},
number = ,
volume = ,
place = {United States},
year = {1989},
month = {10}
}

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