title: Purification and X-ray crystallographic analysis of 7-keto-8-aminopelargonic acid (KAPA) synthase from Mycobacterium smegmatis.
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 VM value of 2.06 Å(3) Da(-1) and a solvent content of 40.20%.
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 27 Shanda Nan Road, Jinan, Shandong 250100, People's Republic of China.
authors.forename: Shanghua
authors.initials: S
authors.lastname: Fan
authors.affiliation: National Laboratory of Biomacromolecules and Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
authors.forename: Defeng
authors.initials: D
authors.lastname: Li
authors.affiliation: National Laboratory of Biomacromolecules and Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
authors.forename: Joy
authors.initials: J
authors.lastname: Fleming
authors.affiliation: National Laboratory of Biomacromolecules and Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Hong
authors.affiliation: Center for Tuberculosis Control of Guangdong Province, Guangzhou, People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.affiliation: Center for Tuberculosis Control of Guangdong Province, Guangzhou, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhou
authors.affiliation: National Laboratory of Biomacromolecules and Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Bi
authors.affiliation: National Laboratory of Biomacromolecules and Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
authors.forename: Dacheng
authors.initials: D
authors.lastname: Wang
authors.affiliation: National Laboratory of Biomacromolecules and Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
authors.forename: Xianen
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 27 Shanda Nan Road, Jinan, Shandong 250100, People's Republic of China.
authors.forename: Guanjun
authors.initials: G
authors.lastname: Chen
mesh_terms: D000217:Acyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D003460:Crystallization; D018360:Crystallography, X-Ray; D008969:Molecular Sequence Data; D020102:Mycobacterium smegmatis / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D000217:Acyltransferases; C021929:8-amino-7-oxononanoate synthase
keywords: 7-keto-8-aminopelargonic acid (KAPA) synthase; Mycobacterium smegmatis; biotin-synthesis pathway