Purification and X-ray crystallographic analysis of 7-keto-8-aminopelargonic acid (KAPA) synthase from Mycobacterium smegmatis

Shanghua Fan, Defeng Li, Joy Fleming, Yuan Hong, Tao Chen, Lin Zhou, Lijun Bi, Dacheng Wang, Xianen Zhang, Guanjun Chen

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

7-Keto-8-aminopelargonic acid synthase (KAPA synthase; BioF) is an essential enzyme for mycobacterial growth that catalyses the first committed step in the biotin-synthesis pathway. It is a pyridoxal 5′-phosphate (PLP)-dependent enzyme and is a potential drug target. Here, the cloning, expression, purification and crystallization of KAPA synthase from Mycobacterium smegmatis (MsBioF) and the characterization of MsBioF crystals using X-ray diffraction are described. The crystals diffracted to 2.3 Å resolution and belonged to the monoclinic space group P21, with unit-cell parameters a = 70.88, b = 91.68, c = 109.84 Å, β = 97.8°. According to the molecular weight of MsBioF, the unit-cell parameters and the self-rotation function map, four molecules are present in each asymmetric unit with a V M value of 2.06 Å3 Da-1 and a solvent content of 40.20%.

Original languageEnglish (US)
Pages (from-to)1372-1375
Number of pages4
JournalActa Crystallographica Section F:Structural Biology Communications
Volume70
DOIs
StatePublished - Jan 1 2014
Externally publishedYes

Fingerprint

8-amino-7-oxononanoate synthase
Mycobacterium smegmatis
purification
Purification
X-Rays
X rays
acids
enzymes
Crystals
biotin
Pyridoxal Phosphate
x rays
Cloning
Enzymes
Biotin
Crystallization
cells
X-Ray Diffraction
crystals
Organism Cloning

Keywords

  • 7-keto-8-aminopelargonic acid (KAPA) synthase
  • biotin-synthesis pathway
  • Mycobacterium smegmatis

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

Cite this

Purification and X-ray crystallographic analysis of 7-keto-8-aminopelargonic acid (KAPA) synthase from Mycobacterium smegmatis. / Fan, Shanghua; Li, Defeng; Fleming, Joy; Hong, Yuan; Chen, Tao; Zhou, Lin; Bi, Lijun; Wang, Dacheng; Zhang, Xianen; Chen, Guanjun.

In: Acta Crystallographica Section F:Structural Biology Communications, Vol. 70, 01.01.2014, p. 1372-1375.

Research output: Contribution to journalArticle

Fan, Shanghua ; Li, Defeng ; Fleming, Joy ; Hong, Yuan ; Chen, Tao ; Zhou, Lin ; Bi, Lijun ; Wang, Dacheng ; Zhang, Xianen ; Chen, Guanjun. / Purification and X-ray crystallographic analysis of 7-keto-8-aminopelargonic acid (KAPA) synthase from Mycobacterium smegmatis. In: Acta Crystallographica Section F:Structural Biology Communications. 2014 ; Vol. 70. pp. 1372-1375.
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AU - Chen, Tao

AU - Zhou, Lin

AU - Bi, Lijun

AU - Wang, Dacheng

AU - Zhang, Xianen

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