title: Purification and crystallographic analysis of a FAD-dependent halogenase from Streptomyces sp. JCM9888.
abstract: A new FAD (flavin adenine dinucleotide)-dependent halogenase HalY from Streptomyces sp. JCM9888 was reported to be involved in the regioselective halogenation of adenine. HalY is a variant B FAD-dependent halogenase that is most similar to the halogenase PltA involved in pyoluteorin biosynthesis. This study reports the overexpression and purification of HalY with an N-terminal hexahistidine tag, followed by crystallization experiments and X-ray crystallographic analysis. HalY was purified as a monomer in solution and crystallized to give X-ray diffraction to a resolution of 1.7 Å. The crystal belonged to the monoclinic space group P21, with unit-cell parameters a = 41.4, b = 113.4, c = 47.6 Å, α = γ = 90, β = 107.4°, and contained one monomer of HalY in the asymmetric unit, with a calculated Matthews coefficient of 2.3 Å(3) Da(-1) and a solvent content of 46%. The structure of the halogenase CndH was used as a search model in molecular replacement to obtain the initial model of HalY. Manual model building and structure refinement of HalY are in progress.
authors.affiliation: College of Life Science, Key Laboratory of Bio-Resources and Eco-Environment of the Ministry of Education, Center for Growth, Metabolism and Aging, Sichuan University, Chengdu 610064, People's Republic of China.
authors.forename: Yanqun
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: College of Life Science, Key Laboratory of Bio-Resources and Eco-Environment of the Ministry of Education, Center for Growth, Metabolism and Aging, Sichuan University, Chengdu 610064, People's Republic of China.
authors.forename: Baohua
authors.initials: B
authors.lastname: Yan
authors.affiliation: College of Life Science, Key Laboratory of Bio-Resources and Eco-Environment of the Ministry of Education, Center for Growth, Metabolism and Aging, Sichuan University, Chengdu 610064, People's Republic of China.
authors.forename: Ting
authors.initials: T
authors.lastname: Yang
authors.affiliation: College of Life Science, Key Laboratory of Bio-Resources and Eco-Environment of the Ministry of Education, Center for Growth, Metabolism and Aging, Sichuan University, Chengdu 610064, People's Republic of China.
authors.forename: Jian
authors.initials: J
authors.lastname: Jiang
authors.affiliation: College of Life Science, Key Laboratory of Bio-Resources and Eco-Environment of the Ministry of Education, Center for Growth, Metabolism and Aging, Sichuan University, Chengdu 610064, People's Republic of China.
authors.forename: Heng
authors.initials: H
authors.lastname: Wei
authors.affiliation: College of Life Science, Key Laboratory of Bio-Resources and Eco-Environment of the Ministry of Education, Center for Growth, Metabolism and Aging, Sichuan University, Chengdu 610064, People's Republic of China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Zhu
mesh_terms: D000225:Adenine / Q000737:chemistry; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry*; D015870:Gene Expression; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000737:chemistry* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000737:chemistry* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D005182:Flavin-Adenine Dinucleotide; D010088:Oxidoreductases; D000225:Adenine
keywords: FAD-dependent halogenase; adenine; regioselective halogenation