Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1980 Apr;17(4):663–668. doi: 10.1128/aac.17.4.663

Identification of a nicotinamide adenine dinucleotide glycohydrolase and an associated inhibitor in isoniazid-susceptible and -resistant Mycobacterium phlei.

W B Davis
PMCID: PMC283849  PMID: 6249194

Abstract

Nicotinamide adenine dinucleotide glycohydrolase (NADase) activity was demonstrated in the catalases fraction of Sephadex G-200-chromatographed sonic extracts of isoniazid (INH)-susceptible (Inhs) and -resistant (Inhr) Mycobacterium phlei. Since crude extracts had no demonstrable activity even after heating, active fractions of the NADase were purified chromatographically by removing the inhibitor with Sephadex G-200. Assays for oxidized nicotinamide adenine dinucleotide (NAD+) hydrolytic activity were done by following the disappearance of NAD+ by the methods of alcohol dehydrogenase or cyanide addition. The NADase activity was linear with respect to time as well as concentration of enzyme and was inhibited in the presence of 0.04 M NADP, benzoic acid hydrazide, or nicotinamide. Crude extracts or pooled concentrated Sephadex G-200 fractions eluting after the catalase inhibited NADase activity by at least 70%. Inhibitor activity was present in both the Inhs and Inhr strains of M. phlei. The activity of the partially purified inhibitors was reversible by INH or nicotinic acid hydrazide at levels between 10 and 100 mM. These findings indicate that an NADase inhibitor system which is sensitive to reversal by INH functions in both the Inhs and Inhr strains; however, unlike previous studies with other mycobacterial species, the enzyme is sensitive to inhibition by nicotinamide. Furthermore, the inhibitors are heat stable and sensitive to reversal by nicotinic acid hydrazide as well as INH.

Full text

PDF
663

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bekierkunst A. Nicotinamide-adenine dinucleotide in tubercle bacilli exposed to isoniazid. Science. 1966 Apr 22;152(3721):525–526. doi: 10.1126/science.152.3721.525. [DOI] [PubMed] [Google Scholar]
  2. Davis W. B., Phillips D. M. Differentiation of catalases in Mycobacterium phlei on the basis of susceptibility to isoniazid: association with peroxidase and acquired resistance to isoniazid. Antimicrob Agents Chemother. 1977 Oct;12(4):529–533. doi: 10.1128/aac.12.4.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Devi B. G., Shaila M. S., Ramakrishnan T., Gopinathan K. P. The purification and properties of peroxidase in Mycobacterium tuberculosis H37Rv and its possible role in the mechanism of action of isonicotinic acid hydrazide. Biochem J. 1975 Jul;149(1):187–197. doi: 10.1042/bj1490187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Diaz G. A., Wayne L. G. Isolation and characterization of catalase produced by Mycobacterium tuberculosis. Am Rev Respir Dis. 1974 Sep;110(3):312–319. doi: 10.1164/arrd.1974.110.3.312. [DOI] [PubMed] [Google Scholar]
  5. GOPINATHAN K. P., SIRSI M., RAMAKRISHNAN T. Nicotin-amide-adenine nucleotides of Mycobacterium tuberculosis H37Rv. Biochem J. 1963 May;87:444–448. doi: 10.1042/bj0870444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gopinathan K. P., Sirsi M., Vaidyanathan C. S. Nicotinamide-adenine dinucleotide glycohydrolase of Mycobacterium tuberculosis H37Rv. Biochem J. 1964 May;91(2):277–282. doi: 10.1042/bj0910277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jackett P. S., Aber V. R., Mitchison D. A. The relationship between nicotinamide adenine dinucleotide concentration and antibacterial activity of isoniazid in Mycobacterium tuberculosis. Am Rev Respir Dis. 1977 Apr;115(4):601–607. doi: 10.1164/arrd.1977.115.4.601. [DOI] [PubMed] [Google Scholar]
  8. KERN M., NATALE R. A diphosphopyridine nucleotidase and its protein inhibitor from Mycobacterium butyricum. J Biol Chem. 1958 Mar;231(1):41–51. [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. MIDDLEBROOK G. Isoniazid-resistance and catalase activity of tubercle bacilli; a preliminary report. Am Rev Tuberc. 1954 Mar;69(3):471–472. doi: 10.1164/art.1954.69.3.471. [DOI] [PubMed] [Google Scholar]
  11. PELLETIER O., CAMPBELL J. A. Direct determination of niacinamide in multivitamin preparations. J Pharm Sci. 1961 Nov;50:926–928. doi: 10.1002/jps.2600501109. [DOI] [PubMed] [Google Scholar]
  12. Sriprakash K. S., Ramakrishnan T. Isoniazid-resistant mutants of Mycobacterium tuberculosis H37RV: uptake of isoniazid and the properties of NADase inhibitor. J Gen Microbiol. 1970 Jan;60(1):125–132. doi: 10.1099/00221287-60-1-125. [DOI] [PubMed] [Google Scholar]
  13. Takayama K., Schnoes H. K., Armstrong E. L., Boyle R. W. Site of inhibitory action of isoniazid in the synthesis of mycolic acids in Mycobacterium tuberculosis. J Lipid Res. 1975 Jul;16(4):308–317. [PubMed] [Google Scholar]
  14. Wang L., Takayama K. Relationship between the uptake of isoniazid and its action on in vivo mycolic acid synthesis in Mycobacterium tuberculosis. Antimicrob Agents Chemother. 1972 Dec;2(6):438–441. doi: 10.1128/aac.2.6.438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Winder F. G., Collins P. B. Inhibition by isoniazid of synthesis of mycolic acids in Mycobacterium tuberculosis. J Gen Microbiol. 1970 Sep;63(1):41–48. doi: 10.1099/00221287-63-1-41. [DOI] [PubMed] [Google Scholar]
  16. YONEDA M., ASANO N. Competitive action of isonicotinic acid hydrazide and pyridoxal in the amino acid decarboxylation of Escherichia coli. Science. 1953 Mar 13;117(3037):277–279. doi: 10.1126/science.117.3037.277. [DOI] [PubMed] [Google Scholar]
  17. Youatt J., Tham S. H. An enzyme system of Mycobacterium tuberculosis that reacts specifically with isoniazid. I. Am Rev Respir Dis. 1969 Jul;100(1):25–30. doi: 10.1164/arrd.1969.100.1.25. [DOI] [PubMed] [Google Scholar]
  18. ZATMAN L. J., KAPLAN N. O., COLOWICK S. P., CIOTTI M. M. Effect of isonicotinic acid hydrazide on diphosphopyridine nucleotidases. J Biol Chem. 1954 Aug;209(2):453–466. [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES