Characterization of Mycobacterium smegmatis expressing the Mycobacterium tuberculosis fatty acid synthase I (fas1) gene

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Unlike most other bacteria, mycobacteria make fatty acids with the multidomain enzyme eukaryote-like fatty acid synthase I (FASI). Previous studies have demonstrated that the tuberculosis drug pyrazinamide and 5-chloro-pyrazinamide target FASI activity. Biochemical studies have revealed that in addition to C16:0, Mycobacterium tuberculosis FASI synthesizes C26:0 fatty acid, while the Mycobacterium smegmatis enzyme makes C24:0 fatty acid. In order to express M. tuberculosis FASI in a rapidly growing Mycobacterium and to characterize the M. tuberculosis FASI in vivo, we constructed an M. smegmatis Δfas1 strain which contained the M. tuberculosis fas1 homologue. The M. smegmatis Δfas1 (attB::M. tuberculosis fas1) strain grew more slowly than the parental M. smegmatis strain and was more susceptible to 5-chloro-pyrazinamide. Surprisingly, while the M. smegmatis Δfas1 (attB::M. tuberculosis fas1) strain produced C 26:0, it predominantly produced C24:0. These results suggest that the fatty acid elongation that produces C24:0 or C 26:0 in vivo is due to a complex interaction among FASI, FabH, and FASII and possibly other systems and is not solely due to FASI elongation, as previously suggested by in vitro studies.

Original languageEnglish (US)
Pages (from-to)4051-4055
Number of pages5
JournalJournal of Bacteriology
Volume186
Issue number13
DOIs
StatePublished - Jul 1 2004

Fingerprint

Mycobacterium smegmatis
Fatty Acid Synthases
Glycogen Synthase
Mycobacterium tuberculosis
Pyrazinamide
Fatty Acids
Genes
Mycobacterium
Enzymes
Eukaryota
Tuberculosis
mycobacteria fatty acid synthase I
Bacteria
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

@article{ac29589a92604e24b4bbfadfc75d54d5,
title = "Characterization of Mycobacterium smegmatis expressing the Mycobacterium tuberculosis fatty acid synthase I (fas1) gene",
abstract = "Unlike most other bacteria, mycobacteria make fatty acids with the multidomain enzyme eukaryote-like fatty acid synthase I (FASI). Previous studies have demonstrated that the tuberculosis drug pyrazinamide and 5-chloro-pyrazinamide target FASI activity. Biochemical studies have revealed that in addition to C16:0, Mycobacterium tuberculosis FASI synthesizes C26:0 fatty acid, while the Mycobacterium smegmatis enzyme makes C24:0 fatty acid. In order to express M. tuberculosis FASI in a rapidly growing Mycobacterium and to characterize the M. tuberculosis FASI in vivo, we constructed an M. smegmatis Δfas1 strain which contained the M. tuberculosis fas1 homologue. The M. smegmatis Δfas1 (attB::M. tuberculosis fas1) strain grew more slowly than the parental M. smegmatis strain and was more susceptible to 5-chloro-pyrazinamide. Surprisingly, while the M. smegmatis Δfas1 (attB::M. tuberculosis fas1) strain produced C 26:0, it predominantly produced C24:0. These results suggest that the fatty acid elongation that produces C24:0 or C 26:0 in vivo is due to a complex interaction among FASI, FabH, and FASII and possibly other systems and is not solely due to FASI elongation, as previously suggested by in vitro studies.",
author = "Oren Zimhony and Catherine Vilch{\`e}ze and Jacobs, {William R.}",
year = "2004",
month = "7",
day = "1",
doi = "10.1128/JB.186.13.4051-4055.2004",
language = "English (US)",
volume = "186",
pages = "4051--4055",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "13",

}

TY - JOUR

T1 - Characterization of Mycobacterium smegmatis expressing the Mycobacterium tuberculosis fatty acid synthase I (fas1) gene

AU - Zimhony, Oren

AU - Vilchèze, Catherine

AU - Jacobs, William R.

PY - 2004/7/1

Y1 - 2004/7/1

N2 - Unlike most other bacteria, mycobacteria make fatty acids with the multidomain enzyme eukaryote-like fatty acid synthase I (FASI). Previous studies have demonstrated that the tuberculosis drug pyrazinamide and 5-chloro-pyrazinamide target FASI activity. Biochemical studies have revealed that in addition to C16:0, Mycobacterium tuberculosis FASI synthesizes C26:0 fatty acid, while the Mycobacterium smegmatis enzyme makes C24:0 fatty acid. In order to express M. tuberculosis FASI in a rapidly growing Mycobacterium and to characterize the M. tuberculosis FASI in vivo, we constructed an M. smegmatis Δfas1 strain which contained the M. tuberculosis fas1 homologue. The M. smegmatis Δfas1 (attB::M. tuberculosis fas1) strain grew more slowly than the parental M. smegmatis strain and was more susceptible to 5-chloro-pyrazinamide. Surprisingly, while the M. smegmatis Δfas1 (attB::M. tuberculosis fas1) strain produced C 26:0, it predominantly produced C24:0. These results suggest that the fatty acid elongation that produces C24:0 or C 26:0 in vivo is due to a complex interaction among FASI, FabH, and FASII and possibly other systems and is not solely due to FASI elongation, as previously suggested by in vitro studies.

AB - Unlike most other bacteria, mycobacteria make fatty acids with the multidomain enzyme eukaryote-like fatty acid synthase I (FASI). Previous studies have demonstrated that the tuberculosis drug pyrazinamide and 5-chloro-pyrazinamide target FASI activity. Biochemical studies have revealed that in addition to C16:0, Mycobacterium tuberculosis FASI synthesizes C26:0 fatty acid, while the Mycobacterium smegmatis enzyme makes C24:0 fatty acid. In order to express M. tuberculosis FASI in a rapidly growing Mycobacterium and to characterize the M. tuberculosis FASI in vivo, we constructed an M. smegmatis Δfas1 strain which contained the M. tuberculosis fas1 homologue. The M. smegmatis Δfas1 (attB::M. tuberculosis fas1) strain grew more slowly than the parental M. smegmatis strain and was more susceptible to 5-chloro-pyrazinamide. Surprisingly, while the M. smegmatis Δfas1 (attB::M. tuberculosis fas1) strain produced C 26:0, it predominantly produced C24:0. These results suggest that the fatty acid elongation that produces C24:0 or C 26:0 in vivo is due to a complex interaction among FASI, FabH, and FASII and possibly other systems and is not solely due to FASI elongation, as previously suggested by in vitro studies.

UR - http://www.scopus.com/inward/record.url?scp=3042606702&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3042606702&partnerID=8YFLogxK

U2 - 10.1128/JB.186.13.4051-4055.2004

DO - 10.1128/JB.186.13.4051-4055.2004

M3 - Article

C2 - 15205406

AN - SCOPUS:3042606702

VL - 186

SP - 4051

EP - 4055

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 13

ER -