Abstract
Intracellular levels of three coenzyme A (CoA) molecular species, i.e., nonesterified CoA (CoASH), acetyl-CoA, and malonyl-CoA, in a variety of aerobic and facultatively anaerobic bacteria were analyzed by the acyl-CoA cycling method developed by us. It was demonstrated that there was an intrinsic difference between aerobes and facultative anaerobes in the changes in the size and composition of CoA pools. The CoA pools in the aerobic bacteria hardly changed and were significantly smaller than those of the facultatively anaerobic bacteria. On the other hand, in the facultatively anaerobic bacteria, the size and composition of the CoA pool drastically changed within minutes in response to the carbon and energy source provided. Acetyl-CoA was the major component of the CoA pool in the facultative anaerobes grown on sufficient glucose, although CoASH was dominant in the aerobes. Therefore, the acetyl-CoA/CoASH ratios in facultatively anaerobic bacteria were 10 times higher than those in aerobic bacteria. In Escherichia coli K-12 cells, the addition of reagents to inhibit the respiratory system led to a rapid decrease in the amount of acetyl-CoA with a concomitant increase in the amount of CoASH, whereas the addition of cerulenin, a specific inhibitor of fatty acid synthase, triggered the intracellular accumulation of malonyl-CoA. The acylation and deacylation of the three CoA molecular species coordinated with the energy-yielding systems and the restriction of the fatty acid-synthesizing system of cells. These data suggest that neither the accumulation of acetyl-CoA nor that of malonyl-CoA exerts negative feedback on pyruvate dehydrogenase and acetyl-CoA carboxylase, respectively.
Full Text
The Full Text of this article is available as a PDF (332.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Andersen K. B., von Meyenburg K. Charges of nicotinamide adenine nucleotides and adenylate energy charge as regulatory parameters of the metabolism in Escherichia coli. J Biol Chem. 1977 Jun 25;252(12):4151–4156. [PubMed] [Google Scholar]
- Atkinson D. E. Biological feedback control at the molecular level. Science. 1965 Nov 12;150(3698):851–857. doi: 10.1126/science.150.3698.851. [DOI] [PubMed] [Google Scholar]
- Atkinson D. E., Walton G. M. Adenosine triphosphate conservation in metabolic regulation. Rat liver citrate cleavage enzyme. J Biol Chem. 1967 Jul 10;242(13):3239–3241. [PubMed] [Google Scholar]
- BROWN G. M. The metabolism of pantothenic acid. J Biol Chem. 1959 Feb;234(2):370–378. [PubMed] [Google Scholar]
- Ball W. J., Jr, Atkinson D. E. Adenylate energy charge in Saccharomyces cerevisiae during starvation. J Bacteriol. 1975 Mar;121(3):975–982. doi: 10.1128/jb.121.3.975-982.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bochner B. R., Ames B. N. Complete analysis of cellular nucleotides by two-dimensional thin layer chromatography. J Biol Chem. 1982 Aug 25;257(16):9759–9769. [PubMed] [Google Scholar]
- Boynton Z. L., Bennett G. N., Rudolph F. B. Intracellular Concentrations of Coenzyme A and Its Derivatives from Clostridium acetobutylicum ATCC 824 and Their Roles in Enzyme Regulation. Appl Environ Microbiol. 1994 Jan;60(1):39–44. doi: 10.1128/aem.60.1.39-44.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chapman A. G., Atkinson D. E. Adenine nucleotide concentrations and turnover rates. Their correlation with biological activity in bacteria and yeast. Adv Microb Physiol. 1977;15:253–306. doi: 10.1016/s0065-2911(08)60318-5. [DOI] [PubMed] [Google Scholar]
- Chapman A. G., Fall L., Atkinson D. E. Adenylate energy charge in Escherichia coli during growth and starvation. J Bacteriol. 1971 Dec;108(3):1072–1086. doi: 10.1128/jb.108.3.1072-1086.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- D'Agnolo G., Rosenfeld I. S., Awaya J., Omura S., Vagelos P. R. Inhibition of fatty acid synthesis by the antibiotic cerulenin. Specific inactivation of beta-ketoacyl-acyl carrier protein synthetase. Biochim Biophys Acta. 1973 Nov 29;326(2):155–156. doi: 10.1016/0005-2760(73)90241-5. [DOI] [PubMed] [Google Scholar]
- Furukawa H., Tsay J. T., Jackowski S., Takamura Y., Rock C. O. Thiolactomycin resistance in Escherichia coli is associated with the multidrug resistance efflux pump encoded by emrAB. J Bacteriol. 1993 Jun;175(12):3723–3729. doi: 10.1128/jb.175.12.3723-3729.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayashi T., Yamamoto O., Sasaki H., Kawaguchi A., Okazaki H. Mechanism of action of the antibiotic thiolactomycin inhibition of fatty acid synthesis of Escherichia coli. Biochem Biophys Res Commun. 1983 Sep 30;115(3):1108–1113. doi: 10.1016/s0006-291x(83)80050-3. [DOI] [PubMed] [Google Scholar]
- Hayashi T., Yamamoto O., Sasaki H., Okazaki H., Kawaguchi A. Inhibition of fatty acid synthesis by the antibiotic thiolactomycin. J Antibiot (Tokyo) 1984 Nov;37(11):1456–1461. doi: 10.7164/antibiotics.37.1456. [DOI] [PubMed] [Google Scholar]
- Jackowski S., Murphy C. M., Cronan J. E., Jr, Rock C. O. Acetoacetyl-acyl carrier protein synthase. A target for the antibiotic thiolactomycin. J Biol Chem. 1989 May 5;264(13):7624–7629. [PubMed] [Google Scholar]
- Jackowski S., Rock C. O. Consequences of reduced intracellular coenzyme A content in Escherichia coli. J Bacteriol. 1986 Jun;166(3):866–871. doi: 10.1128/jb.166.3.866-871.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackowski S., Rock C. O. Metabolism of 4'-phosphopantetheine in Escherichia coli. J Bacteriol. 1984 Apr;158(1):115–120. doi: 10.1128/jb.158.1.115-120.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackowski S., Rock C. O. Regulation of coenzyme A biosynthesis. J Bacteriol. 1981 Dec;148(3):926–932. doi: 10.1128/jb.148.3.926-932.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karl D. M. Cellular nucleotide measurements and applications in microbial ecology. Microbiol Rev. 1980 Dec;44(4):739–796. doi: 10.1128/mr.44.4.739-796.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kauppinen S., Siggaard-Andersen M., von Wettstein-Knowles P. beta-Ketoacyl-ACP synthase I of Escherichia coli: nucleotide sequence of the fabB gene and identification of the cerulenin binding residue. Carlsberg Res Commun. 1988;53(6):357–370. doi: 10.1007/BF02983311. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., PASSONNEAU J. V., SCHULZ D. W., ROCK M. K. The measurement of pyridine nucleotides by enzymatic cycling. J Biol Chem. 1961 Oct;236:2746–2755. [PubMed] [Google Scholar]
- Lomovskaya O., Lewis K. Emr, an Escherichia coli locus for multidrug resistance. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8938–8942. doi: 10.1073/pnas.89.19.8938. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MAKMAN R. S., SUTHERLAND E. W. ADENOSINE 3',5'-PHOSPHATE IN ESCHERICHIA COLI. J Biol Chem. 1965 Mar;240:1309–1314. [PubMed] [Google Scholar]
- Miyakawa S., Suzuki K., Noto T., Harada Y., Okazaki H. Thiolactomycin, a new antibiotic. IV. Biological properties and chemotherapeutic activity in mice. J Antibiot (Tokyo) 1982 Apr;35(4):411–419. doi: 10.7164/antibiotics.35.411. [DOI] [PubMed] [Google Scholar]
- Nishida I., Kawaguchi A., Yamada M. Effect of thiolactomycin on the individual enzymes of the fatty acid synthase system in Escherichia coli. J Biochem. 1986 May;99(5):1447–1454. doi: 10.1093/oxfordjournals.jbchem.a135614. [DOI] [PubMed] [Google Scholar]
- Noto T., Miyakawa S., Oishi H., Endo H., Okazaki H. Thiolactomycin, a new antibiotic. III. In vitro antibacterial activity. J Antibiot (Tokyo) 1982 Apr;35(4):401–410. doi: 10.7164/antibiotics.35.401. [DOI] [PubMed] [Google Scholar]
- Oishi H., Noto T., Sasaki H., Suzuki K., Hayashi T., Okazaki H., Ando K., Sawada M. Thiolactomycin, a new antibiotic. I. Taxonomy of the producing organism, fermentation and biological properties. J Antibiot (Tokyo) 1982 Apr;35(4):391–395. doi: 10.7164/antibiotics.35.391. [DOI] [PubMed] [Google Scholar]
- Omura S. The antibiotic cerulenin, a novel tool for biochemistry as an inhibitor of fatty acid synthesis. Bacteriol Rev. 1976 Sep;40(3):681–697. doi: 10.1128/br.40.3.681-697.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwartz E. R., Old L. O., Reed L. J. Regulatory properties of pyruvate dehydrogenase from Escherichia coli. Biochem Biophys Res Commun. 1968 May 10;31(3):495–500. doi: 10.1016/0006-291x(68)90504-4. [DOI] [PubMed] [Google Scholar]
- Song W. J., Jackowski S. Cloning, sequencing, and expression of the pantothenate kinase (coaA) gene of Escherichia coli. J Bacteriol. 1992 Oct;174(20):6411–6417. doi: 10.1128/jb.174.20.6411-6417.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Takamura Y., Nomura G. Changes in the intracellular concentration of acetyl-CoA and malonyl-CoA in relation to the carbon and energy metabolism of Escherichia coli K12. J Gen Microbiol. 1988 Aug;134(8):2249–2253. doi: 10.1099/00221287-134-8-2249. [DOI] [PubMed] [Google Scholar]
- Tsay J. T., Rock C. O., Jackowski S. Overproduction of beta-ketoacyl-acyl carrier protein synthase I imparts thiolactomycin resistance to Escherichia coli K-12. J Bacteriol. 1992 Jan;174(2):508–513. doi: 10.1128/jb.174.2.508-513.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vallari D. S., Jackowski S. Biosynthesis and degradation both contribute to the regulation of coenzyme A content in Escherichia coli. J Bacteriol. 1988 Sep;170(9):3961–3966. doi: 10.1128/jb.170.9.3961-3966.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vallari D. S., Jackowski S., Rock C. O. Regulation of pantothenate kinase by coenzyme A and its thioesters. J Biol Chem. 1987 Feb 25;262(6):2468–2471. [PubMed] [Google Scholar]
- Vance D., Goldberg I., Mitsuhashi O., Bloch K. Inhibition of fatty acid synthetases by the antibiotic cerulenin. Biochem Biophys Res Commun. 1972 Aug 7;48(3):649–656. doi: 10.1016/0006-291x(72)90397-x. [DOI] [PubMed] [Google Scholar]
- Winkler H. H., Wilson T. H. The role of energy coupling in the transport of beta-galactosides by Escherichia coli. J Biol Chem. 1966 May 25;241(10):2200–2211. [PubMed] [Google Scholar]
- Wright J. A., Maeba P., Sanwal B. D. Allosteric regulation of the activity of citrate snythetase of Escherichia coli by alpha-ketoglutarate. Biochem Biophys Res Commun. 1967 Oct 11;29(1):34–38. doi: 10.1016/0006-291x(67)90536-0. [DOI] [PubMed] [Google Scholar]
- de Mendoza D., Klages Ulrich A., Cronan J. E., Jr Thermal regulation of membrane fluidity in Escherichia coli. Effects of overproduction of beta-ketoacyl-acyl carrier protein synthase I. J Biol Chem. 1983 Feb 25;258(4):2098–2101. [PubMed] [Google Scholar]