Fluorescence detection of metabolic activity of the fatty acid beta oxidation pathway in living cells

2020 ◽  
Vol 56 (20) ◽  
pp. 3023-3026
Author(s):  
Shohei Uchinomiya ◽  
Naoya Matsunaga ◽  
Koichiro Kamoda ◽  
Ryosuke Kawagoe ◽  
Akito Tsuruta ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Fluorescent Probe ◽  
Fluorescence Imaging ◽  
Fluorescence Detection ◽  
Metabolic Activity ◽  
Living Cells ◽  
Beta Oxidation ◽  
Enzyme Reactions ◽  
Oxidation Pathway ◽  
Fatty Acid Beta Oxidation

Fluorescence imaging of fatty acid beta oxidation (FAO) with a fluorescent probe metabolically degraded by sequential enzyme reactions of FAO.

BIOSYNTHESIS OF BUTYRIC ACID FROM GLUCOSE THROUGH THE INVERTED FATTY ACID BETA-OXIDATION PATHWAY BY RECOMBINANT ESCHERICHIA COLI STRAIN IN BIOCATALYTIC MODE DUE TO ENFORCED ATP HYDROLYSIS

2021 ◽  
Vol 1 (19) ◽  
pp. 268-269
Author(s):  
A.Yu. Skorokhodova ◽  
V.G. Debabov
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Butyric Acid ◽  
Atp Hydrolysis ◽  
Coli Strain ◽  
Recombinant Escherichia Coli ◽  
Beta Oxidation ◽  
Oxidation Pathway ◽  
Full Cell ◽  
Fatty Acid Beta Oxidation

The feasibility of the application of enforced ATP hydrolysis to ensure anaerobic functioning of Escherichia coli strain producing butyric acid through the inverted fatty acid beta-oxidation pathway as a full-cell biocatalyst has been demonstrated.

BIOSYNTHESIS OF 3-HYDROXYFUNCTIONALIZED C4-C8 ALIPHATIC CARBOXYLIC ACID FROM GLUCOSE THROUGH INVERTED FATTY ACID BETA- OXIDATION PATHWAY BY RECOMBINANT ESCHERICHIA COLI STRAINS

2020 ◽  
pp. 10-11
Author(s):  
A. Skorokhodova ◽  
V. Debabov
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Carboxylic Acid ◽  
Recombinant Escherichia Coli ◽  
Beta Oxidation ◽  
Aliphatic Carboxylic Acid ◽  
Oxidation Pathway ◽  
Fatty Acid Beta Oxidation

The feasibility of the biosynthesis from glucose of 3-hydroxyfunctionalized C4-C8 carboxylates upon the reversal of the fatty acid beta-oxidation in recombinant Escherichia coli strains has been demonstrated.

Study of the Potential of Reversal of the Fatty Acid Beta-Oxidation Pathway for Stereoselective Biosynthesis of (S)-1,3-Butanediol from Glucose by Recombinant Escherichia coli Strains

2019 ◽  
Vol 35 (5) ◽  
pp. 12-19
Author(s):  
A.Yu. Skorokhodova ◽  
V.G. Debabov
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Essential Gene ◽  
Coli Strain ◽  
Recombinant Escherichia Coli ◽  
Beta Oxidation ◽  
Oxidation Pathway ◽  
Gene Coding ◽  
Fundamental Research ◽  
Fatty Acid Beta Oxidation

A possible contribution of collateral enzymes to the formation of key precursor metabolite, 3-hydroxybutyryl-CoA, in a recombinant Escherichia coli strain engineered for 1,3-butanediol biosynthesis from glucose through the inverted fatty acid beta-oxidation pathway has been evaluated. The inactivation of the 3-hydroxyadipyl-CoA dehydrogenase gene, paaH, did not prevent the 1,3-butanol biosynthesis during anaerobic glucose utilization by the strain with the intact essential gene fabG coding for 3-ketoacyl-ACP reductase, which can catalyze the conversion of acetoacetyl-CoA to (R)-3-hydroxybutyryl-CoA. The subsequent inactivation in the strain of fadB gene coding for (S)-stereospecific 3-hydroxyacyl-CoA dehydrogenase of the fatty acid beta-oxidation led to the abolishment of the 1,3-butanediol synthesis. The respective diol was also not found among the products secreted by the strain possessing the intact fabG and paaH genes upon an individual deletion of fadB gene. It was established that the collateral enzymes did not participate in the formation of 3-hydroxybutyryl-CoA in the studied strains and the respective CoA-derivative was synthesized solely by the (S)-specific enzyme of the fatty acid beta-oxidation pathway. The obtained results indicate that the reversal of the fatty acid beta-oxidation pathway can ensure the enantioselective biosynthesis of the (S)-stereoisomer of 1,3-butanediol in engineered E. coli strains. 1,3-butanediol, fatty acid beta-oxidation, Escherichia coli, glucose, metabolic engineering, stereoisomer. The work was carried out with financial support Russian Foundation for Fundamental Research (No. 18-29-08059).

scholarly journals Differential protein expression during growth on model and commercial mixtures of naphthenic acids in Pseudomonas fluorescens Pf-5

2021 ◽  
Author(s):  
Boyd McKew ◽  
Richard Johnson ◽  
Lindsey Clothier ◽  
Karl Skeels ◽  
Matthew Ross ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Pseudomonas Fluorescens ◽  
Oil Sands ◽  
Shotgun Proteomics ◽  
Structural Similarity ◽  
Naphthenic Acids ◽  
Acid Oxidation ◽  
Beta Oxidation ◽  
Oxidation Pathway ◽  
Fatty Acid Beta Oxidation

Naphthenic acids (NAs) are carboxylic acids with the formula (CnH2n+ZO2) and are the toxic, persistent constituents of oil sands process-affected waters (OSPW), produced during oil sands extraction. Currently, the proteins and mechanisms involved in NA biodegradation are unknown. Using LC-MS/MS shotgun proteomics, we identified proteins overexpressed during the growth of Pseudomonas fluorescens Pf5 on a model NA (4-n-butylphenyl)-4-butanoic acid (n-BPBA) and commercial NA mixture (Acros). By day 11, >95% of n-BPBA was degraded. With Acros, a 17% reduction in intensity occurred with 10-18 carbon compounds of the Z family -2 to -14 (major NA species in this mixture). A total of 554 proteins (n-BPBA) and 631 proteins (Acros) were overexpressed during growth on NAs; including several transporters (e.g. ABC transporters), suggesting a cellular protective response from NA toxicity. Several proteins associated with fatty acid, lipid and amino acid metabolism were also overexpressed; including acyl-CoA dehydrogenase and acyl-CoA thioesterase II, which catalyze part of the fatty acid beta-oxidation pathway. Indeed, multiple enzymes involved in the fatty acid oxidation pathway were upregulated. Given the presumed structural similarity between alkyl-carboxylic acid side chains and fatty acids, we postulate that P. fluorescens Pf-5 was using existing fatty acid catabolic pathways (among others) during NA degradation.

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