Changes in Size of Intracellular Pools of Coenzyme A and Its Thioesters inEscherichia coliK-12 Cells to Various Carbon Sources and Stresses

ABSTRACT The coenzyme B12-dependent isobutyryl coenzyme A (CoA) mutase (ICM) and methylmalonyl-CoA mutase (MCM) catalyze the isomerization of n-butyryl-CoA to isobutyryl-CoA and of methylmalonyl-CoA to succinyl-CoA, respectively. The influence that both mutases have on the conversion of n- and isobutyryl-CoA to methylmalonyl-CoA and the use of the latter in polyketide biosynthesis have been investigated with the polyether antibiotic (monensin) producer Streptomyces cinnamonensis. Mutants prepared by inserting a hygromycin resistance gene (hygB) into either icmA or mutB, encoding the large subunits of ICM and MCM, respectively, have been characterized. The icmA::hygB mutant was unable to grow on valine or isobutyrate as the sole carbon source but grew normally on butyrate, indicating a key role for ICM in valine and isobutyrate metabolism in minimal medium. ThemutB::hygB mutant was unable to grow on propionate and grew only weakly on butyrate and isobutyrate as sole carbon sources. 13C-labeling experiments show that in both mutants butyrate and acetoacetate may be incorporated into the propionate units in monensin A without cleavage to acetate units. Hence, n-butyryl-CoA may be converted into methylmalonyl-CoA through a carbon skeleton rearrangement for which neither ICM nor MCM alone is essential.

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Multiple Propionyl Coenzyme A-Supplying Pathways for Production of the Bioplastic Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate) in Haloferax mediterranei

Applied and Environmental Microbiology ◽

10.1128/aem.03915-12 ◽

2013 ◽

Vol 79 (9) ◽

pp. 2922-2931 ◽

Cited By ~ 49

Author(s):

Jing Han ◽

Jing Hou ◽

Fan Zhang ◽

Guomin Ai ◽

Ming Li ◽

...

Keyword(s):

Metabolic Flux ◽

Coenzyme A ◽

Molar Fraction ◽

Carbon Sources ◽

Bioinformatic Analysis ◽

Flux Analysis ◽

The Novel ◽

Haloferax Mediterranei ◽

Content Type ◽

First Time

ABSTRACTHaloferax mediterraneiis able to accumulate the bioplastic poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) with more than 10 mol% 3-hydroxyvalerate (3HV) from unrelated carbon sources. However, the pathways that produce propionyl coenzyme A (propionyl-CoA), an important precursor of 3HV monomer, have not yet been determined. Bioinformatic analysis ofH. mediterraneigenome indicated that this strain uses multiple pathways for propionyl-CoA biosynthesis, including the citramalate/2-oxobutyrate pathway, the aspartate/2-oxobutyrate pathway, the methylmalonyl-CoA pathway, and a novel 3-hydroxypropionate pathway. Cofeeding of pathway intermediates and inactivating pathway-specific genes supported that these four pathways were indeed involved in the biosynthesis of 3HV monomer. The novel 3-hydroxypropionate pathway that couples CO2assimilation with PHBV biosynthesis was further confirmed by analysis of13C positional enrichment in 3HV. Notably,13C metabolic flux analysis showed that the citramalate/2-oxobutyrate pathway (53.0% flux) and the 3-hydroxypropionate pathway (30.6% flux) were the two main generators of propionyl-CoA from glucose. In addition, genetic perturbation on the transcriptome of the ΔphaECmutant (deficient in PHBV accumulation) revealed that a considerable number of genes in the four propionyl-CoA synthetic pathways were significantly downregulated. We determined for the first time four propionyl-CoA-supplying pathways for PHBV production in haloarchaea, particularly including a new 3-hydroxypropionate pathway. These results would provide novel strategies for the production of PHBV with controllable 3HV molar fraction.

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Role of Acetyl Coenzyme A Synthesis and Breakdown in Alternative Carbon Source Utilization in Candida albicans

Eukaryotic Cell ◽

10.1128/ec.00253-08 ◽

2008 ◽

Vol 7 (10) ◽

pp. 1733-1741 ◽

Cited By ~ 39

Author(s):

Aaron J. Carman ◽

Slavena Vylkova ◽

Michael C. Lorenz

Keyword(s):

Fatty Acids ◽

Candida Albicans ◽

Coenzyme A ◽

Glyoxylate Cycle ◽

Functional Divergence ◽

Carbon Sources ◽

Growth Defect ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Acetyl Coenzyme

ABSTRACT Acetyl coenzyme A (acetyl-CoA) is the central intermediate of the pathways required to metabolize nonfermentable carbon sources. Three such pathways, i.e., gluconeogenesis, the glyoxylate cycle, and β-oxidation, are required for full virulence in the fungal pathogen Candida albicans. These processes are compartmentalized in the cytosol, mitochondria, and peroxosomes, necessitating transport of intermediates across intracellular membranes. Acetyl-CoA is trafficked in the form of acetate by the carnitine shuttle, and we hypothesized that the enzymes that convert acetyl-CoA to/from acetate, i.e., acetyl-CoA hydrolase (ACH1) and acetyl-CoA synthetase (ACS1 and ACS2), would regulate alternative carbon utilization and virulence. We show that C. albicans strains depleted for ACS2 are unviable in the presence of most carbon sources, including glucose, acetate, and ethanol; these strains metabolize only fatty acids and glycerol, a substantially more severe phenotype than that of Saccharomyces cerevisiae acs2 mutants. In contrast, deletion of ACS1 confers no phenotype, though it is highly induced in the presence of fatty acids, perhaps explaining why acs2 mutants can utilize fatty acids. Strains lacking ACH1 have a mild growth defect on some carbon sources but are fully virulent in a mouse model of disseminated candidiasis. Both ACH1 and ACS2 complement mutations in their S. cerevisiae homolog. Together, these results show that acetyl-CoA metabolism and transport are critical for growth of C. albicans on a wide variety of nutrients. Furthermore, the phenotypic differences between mutations in these highly conserved genes in S. cerevisiae and C. albicans support recent findings that significant functional divergence exists even in fundamental metabolic pathways between these related yeasts.

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Biosynthesis of Isoprenoid Wax Ester in Marinobacter hydrocarbonoclasticus DSM 8798: Identification and Characterization of Isoprenoid Coenzyme A Synthetase and Wax Ester Synthases

Journal of Bacteriology ◽

10.1128/jb.01932-06 ◽

2007 ◽

Vol 189 (10) ◽

pp. 3804-3812 ◽

Cited By ~ 67

Author(s):

Erik Holtzapple ◽

Claudia Schmidt-Dannert

Keyword(s):

Coenzyme A ◽

Draft Genome ◽

Carbon Sources ◽

Wax Ester ◽

Sequence Information ◽

Recombinant Enzymes ◽

Available Carbon ◽

Marinobacter Hydrocarbonoclasticus ◽

Wax Ester Synthase

ABSTRACT Marinobacter hydrocarbonoclasticus DSM 8798 has been reported to synthesize isoprenoid wax ester storage compounds when grown on phytol as the sole carbon source under limiting nitrogen and/or phosphorous conditions. We hypothesized that isoprenoid wax ester synthesis involves (i) activation of an isoprenoid fatty acid by a coenzyme A (CoA) synthetase and (ii) ester bond formation between an isoprenoid alcohol and isoprenoyl-CoA catalyzed, most likely, by an isoprenoid wax ester synthase similar to an acyl wax ester synthase, wax ester synthase/diacylglycerol acyltransferase (WS/DGAT), recently described from Acinetobacter sp. strain ADP1. We used the recently released rough draft genome sequence of a closely related strain, M. aquaeolei VT8, to search for WS/DGAT and acyl-CoA synthetase candidate genes. The sequence information from putative WS/DGAT and acyl-CoA synthetase genes identified in this strain was used to clone homologues from the isoprenoid wax ester synthesizing Marinobacter strain. The activities of the recombinant enzymes were characterized, and two new isoprenoid wax ester synthases capable of synthesizing isoprenoid ester and acyl/isoprenoid hybrid ester in vitro were identified along with an isoprenoid-specific CoA synthetase. One of the Marinobacter wax ester synthases displays several orders of magnitude higher activity toward acyl substrates than any previously characterized acyl-WS and may reflect adaptations to available carbon sources in their environments.

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Levels of coenzyme A – glutathione mixed disulfide in Escherichia coli

Canadian Journal of Biochemistry ◽

10.1139/o78-113 ◽

1978 ◽

Vol 56 (7) ◽

pp. 753-759 ◽

Cited By ~ 6

Author(s):

Peter C. Loewen

Keyword(s):

Escherichia Coli ◽

Coenzyme A ◽

Anaerobic Fermentation ◽

Carbon Sources ◽

Lower Proportion ◽

Fermentable Sugars ◽

Glucose Starvation ◽

Aerobic Growth ◽

Mixed Disulfide ◽

Oxygen Starvation

The pool of coenzyme A – glutathione mixed disulfide (CoASSG) rapidly increased 2.0 times in response to oxygen starvation and 1.5 times in response to glucose starvation but did not change following ammonia starvation. The increase in the CoASSG pool resulted from an increase in the CoASSG fraction of the CoA pool from 42 to 66–93%. Fluoride, cyanide, chloramphenicol, and rifampicin all caused similar increases. Aerobic growth on fermentable sugars resulted in CoASSG making up 40–55% of the CoA pool while growth on nonfermentable carbon sources or anaerobic fermentation resulted in CoASSG replacing acetyl CoA and free CoA to make up 85–95% of the CoA pool. The CoASSG:ATP ratio varied inversely with the growth rate in two groupings of carbon sources made up of either fermentable or nonfermentable molecules. Cultures grown aerobically on fermentable sugars exhibited a lower CoASSG:ATP ratio reflecting the lower proportion of CoASSG in the CoA pool.

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ATP-Citrate Lyase Is Required for Production of Cytosolic Acetyl Coenzyme A and Development in Aspergillus nidulans

Eukaryotic Cell ◽

10.1128/ec.00080-10 ◽

2010 ◽

Vol 9 (7) ◽

pp. 1039-1048 ◽

Cited By ~ 62

Author(s):

Michael J. Hynes ◽

Sandra L. Murray

Keyword(s):

Aspergillus Nidulans ◽

Coenzyme A ◽

Carbon Sources ◽

Gene Arrangement ◽

Atp Citrate Lyase ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Content Type ◽

Acetyl Coenzyme ◽

Citrate Lyase

ABSTRACT Acetyl coenzyme A (CoA) is a central metabolite in carbon and energy metabolism and in the biosynthesis of cellular molecules. A source of cytoplasmic acetyl-CoA is essential for the production of fatty acids and sterols and for protein acetylation, including histone acetylation in the nucleus. In Saccharomyces cerevisiae and Candida albicans acetyl-CoA is produced from acetate by cytoplasmic acetyl-CoA synthetase, while in plants and animals acetyl-CoA is derived from citrate via ATP-citrate lyase. In the filamentous ascomycete Aspergillus nidulans, tandem divergently transcribed genes (aclA and aclB) encode the subunits of ATP-citrate lyase, and we have deleted these genes. Growth is greatly diminished on carbon sources that do not result in cytoplasmic acetyl-CoA, such as glucose and proline, while growth is not affected on carbon sources that result in the production of cytoplasmic acetyl-CoA, such as acetate and ethanol. Addition of acetate restores growth on glucose or proline, and this is dependent on facA, which encodes cytoplasmic acetyl-CoA synthetase, but not on the regulatory gene facB. Transcription of aclA and aclB is repressed by growth on acetate or ethanol. Loss of ATP-citrate lyase results in severe developmental effects, with the production of asexual spores (conidia) being greatly reduced and a complete absence of sexual development. This is in contrast to Sordaria macrospora, in which fruiting body formation is initiated but maturation is defective in an ATP-citrate lyase mutant. Addition of acetate does not repair these defects, indicating a specific requirement for high levels of cytoplasmic acetyl-CoA during differentiation. Complementation in heterokaryons between aclA and aclB deletions for all phenotypes indicates that the tandem gene arrangement is not essential.

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Ceftiofur reduced Fusobacterium leading to uterine microbiota alteration in dairy cows with metritis

Animal Microbiome ◽

10.1186/s42523-021-00077-5 ◽

2021 ◽

Vol 3 (1) ◽

Author(s):

Soo Jin Jeon ◽

Federico Cunha ◽

Rodolfo Daetz ◽

Rodrigo C. Bicalho ◽

Svetlana Lima ◽

...

Keyword(s):

Dairy Cows ◽

Rectal Temperature ◽

Coenzyme A ◽

Carbon Sources ◽

Metabolic Stress ◽

16S Rrna Gene Sequencing ◽

Uterine Disease ◽

Rrna Gene ◽

Hydroxybutyric Acid ◽

Coenzyme A Biosynthesis

Abstract Background Metritis is an inflammatory uterine disease found in ~ 20% of dairy cows after parturition and associated with uterine microbiota with high abundance of Fusobacterium, Bacteroides, and Porphyromonas. Ceftiofur is a common treatment, but the effect on uterine microbiota is poorly understood. Herein, we investigated the short-term impact of ceftiofur on uterine microbiota structure and function in cows with metritis. Eight cows received ceftiofur (CEF) and 10 remained untreated (CON). Uterine swabs were collected for PCR and metagenomic analysis at diagnosis before treatment (5 ± 1 DPP) and 2 days after diagnosis/treatment (7 ± 1 DPP) from the same individuals. Seven CEF and 9 CON passed quality control and were used for 16S rRNA gene sequencing. Results Ceftiofur treatment resulted in uterine microbiota alteration, which was attributed to a decrease in relative abundance of Fusobacterium and in gene contents involved in lipopolysaccharide biosynthesis, whereas uterine microbiota diversity and genes involved in pantothenate and coenzyme A biosynthesis increased. Ceftiofur treatment also reduced rectal temperature and tended to reduce total bacteria in the uterus. However, other uterine pathogens such as Bacteroides and Porphyromonas remained unchanged in CEF. The blaCTX-M gene was detected in 37.5% of metritic cows tested but was not affected by CEF. We found that β-hydroxybutyric acid, pyruvic acid, and L-glutamine were preferentially utilized by Fusobacterium necrophorum according to metabolic activity with 95 carbon sources. Conclusions Ceftiofur treatment leads to alterations in the uterine microbiota that were mainly characterized by reductions in Fusobacterium and genes involved in LPS biosynthesis, which may be associated with a decrease in rectal temperature. The increase in pantothenate and coenzyme A biosynthesis indicates microbial response to metabolic stress caused by ceftiofur. Preference of Fusobacterium for β-hydroxybutyric acid may help to explain why this strain becomes dominant in the uterine microbiota of cows with metritis, and it also may provide a means for development of new therapies for the control of metritis in dairy cows.

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