Open AccessCloning, expression, purification, crystallization and preliminary crystallographic analysis of 5‐aminolaevulinic acid dehydratase from Bacillus subtilis
Author(s) -
Lu Qianda,
Ma Jinming,
Rong Hui,
Fan Jun,
Yuan Ye,
Li Kuai,
Gao Yongxiang,
Zhang Xiao,
Teng Maikun,
Niu Liwen
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110027582
Subject(s) - bacillus subtilis , dehydratase , porphobilinogen , porphobilinogen synthase , escherichia coli , chemistry , biochemistry , crystallization , gene , enzyme , microbiology and biotechnology , crystallography , biology , bacteria , organic chemistry , genetics
5‐Aminolaevulinic acid dehydratase (ALAD), a crucial enzyme in the biosynthesis of tetrapyrrole, catalyses the condensation of two 5‐aminolaevulinic acid (ALA) molecules to form porphobilinogen (PBG). The gene encoding ALAD was amplified from genomic DNA of Bacillus subtilis and the protein was overexpressed in Escherichia coli strain BL21 (DE3). The protein was purified and crystallized with an additional MGSSSSGLVPRGSH– tag at the N‐terminus of the target protein. Diffraction‐quality single crystals were obtained by the hanging‐drop vapour‐diffusion method. An X‐ray diffraction data set was collected at a resolution of 2.7 Å.