The Reconstruction of the Fatty Acid Oxidizing System of Animal Tissues

Beinert, Helmut; Bock, Robert M.; Goldman, Dexter S.; Green, D. E.; Mahler, Henry R.; Mii, Sanae; Stansly, P. G.; Wakil, Salih J.

doi:10.1021/ja01112a536

Cited by 41 publications

(6 citation statements)

References 6 publications

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“…In mutant HK4, the transposon locus was mapped in a gene encoding a putative acetyl-CoA acetyltransferase (EC 2.3.1.16). This acetylCoA acyltransferase belongs to the thiolase family, members of which catalyze the final step in fatty acid beta oxidation (46). In mutant HK6, the transposon insertion was localized 1 bp upstream of a gene coding for a tRNA G18 (ribose-2-O)-methyltransferase, whereas in mutant HK7, it was detected in a gene coding for a polysaccharide deacetyltransferase.…”

Section: Biodegradation Of Dtdb By Available Rhodococcus Sppmentioning

confidence: 99%

Biodegradation of the Organic Disulfide 4,4′-Dithiodibutyric Acid by Rhodococcus spp

Khairy

Wübbeler

Steinbüchel

2015

Appl Environ Microbiol

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c Four Rhodococcus spp. exhibited the ability to use 4,4=-dithiodibutyric acid (DTDB) as a sole carbon source for growth. The most important step for the production of a novel polythioester (PTE) using DTDB as a precursor substrate is the initial cleavage of DTDB. Thus, identification of the enzyme responsible for this step was mandatory. Because Rhodococcus erythropolis strain MI2 serves as a model organism for elucidation of the biodegradation of DTDB, it was used to identify the genes encoding the enzymes involved in DTDB utilization. To identify these genes, transposon mutagenesis of R. erythropolis MI2 was carried out using transposon pTNR-TA. Among 3,261 mutants screened, 8 showed no growth with DTDB as the sole carbon source. In five mutants, the insertion locus was mapped either within a gene coding for a polysaccharide deacetyltransferase, a putative ATPase, or an acetyl coenzyme A transferase, 1 bp upstream of a gene coding for a putative methylase, or 176 bp downstream of a gene coding for a putative kinase. In another mutant, the insertion was localized between genes encoding a putative transcriptional regulator of the TetR family (noxR) and an NADH:flavin oxidoreductase (nox). Moreover, in two other mutants, the insertion loci were mapped within a gene encoding a hypothetical protein in the vicinity of noxR and nox. The interruption mutant generated, R. erythropolis MI2 nox⍀tsr, was unable to grow with DTDB as the sole carbon source. Subsequently, nox was overexpressed and purified, and its activity with DTDB was measured. The specific enzyme activity of Nox amounted to 1.2 ؎ 0.15 U/mg. Therefore, we propose that Nox is responsible for the initial cleavage of DTDB into 2 molecules of 4-mercaptobutyric acid (4MB).T he genus Rhodococcus has attracted great interest in recent years due to the ability of many of its species to degrade and transform a wide range of xenobiotic substances (1). Rhodococci are described as aerobic, Gram-positive, mycolate-containing actinomycetes with high genomic GϩC contents. They have a diverse set of genetic and catabolic features, because of the presence of large linear plasmids in addition to their large chromosomes (2). The number of publications and patents involving Rhodococcus spp. has increased significantly in recent years (3).Rhodococcus spp. have been isolated from a variety of sources. They are ubiquitous in soils contaminated with crude oil and/or other xenobiotic compounds. The ability of Rhodococcus species to degrade substituted hydrocarbons and other chemicals allows them to play a role in the natural degradation and bioremediation of such compounds (4). Many Rhodococcus species are characterized by broad metabolic diversity and an array of unique enzymatic capabilities; therefore, they are also of interest for pharmaceutical, environmental, chemical, and energy studies. They play a significant role in the process of desulfurization of fossil fuels (5) and in the industrial production of acrylamide (6). Strains of Rhodococcus erythropolis have been identi...

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Section: Biodegradation Of Dtdb By Available Rhodococcus Sppmentioning

confidence: 99%

Biodegradation of the Organic Disulfide 4,4′-Dithiodibutyric Acid by Rhodococcus spp

Khairy

Wübbeler

Steinbüchel

2015

Appl Environ Microbiol

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“…p-Ketovaleryl CoA reacts only at 20% of the rate of acetoacetyl CoA; &ketocaproyl-and /3-ketoisocaproyl CoA are hardly attacked. This is in contraat to the broader specificity of crude pig heart enzyme fractions (244) and purified preparations from ox liver (5) and indicates that these preparations contain other thiolase or thiolases for S-pketoacyl CoA derivatives of fatty acid chain length higher than Cr.…”

Section: The Citric Acid Cycle 241mentioning

confidence: 99%

Enzymic Mechanisms in the Citric Acid Cycle

Ochoa¹

1954

Advances in Enzymology - And Related Areas of Molecular Biology

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“…Carbon dioxide production was measured by the method of Claypool and Keefer (2) in which an air stream of known flow is equilibrated with a buffered bromthymol blue solution and the percentage CO2 estimated colorimetrically. The method was modified in that the water-saturated air passing over the fruit was not freed of CO2 and the indicator solution was renewed for each determination.…”

Section: Methodsmentioning

confidence: 99%