Publication: Expression, purification, crystallization and preliminary crystallographic analysis of chitinase A from Vibrio carchariae
Issued Date
2005-12-01
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ISSN
17443091
17443091
17443091
Other identifier(s)
2-s2.0-33744493970
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Mahidol University
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SCOPUS
Bibliographic Citation
Acta Crystallographica Section F: Structural Biology and Crystallization Communications. Vol.61, No.10 (2005), 895-898
Suggested Citation
Chomphunuch Songsiriritthigul, Jirundon Yuvaniyama, Robert C. Robinson, Archara Vongsuwan, Heino Prinz, Wipa Suginta Expression, purification, crystallization and preliminary crystallographic analysis of chitinase A from Vibrio carchariae. Acta Crystallographica Section F: Structural Biology and Crystallization Communications. Vol.61, No.10 (2005), 895-898. doi:10.1107/S1744309105027831 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/16267
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Title
Expression, purification, crystallization and preliminary crystallographic analysis of chitinase A from Vibrio carchariae
Abstract
Chitinase A of Vibrio carchariae was expressed in Escherichia coli M15 host cells as a 575-amino-acid fragment with full enzymatic activity using the pQE60 expression vector. The yield of the highly purified recombinant protein was approximately 70 mg per litre of bacterial culture. The molecular mass of the expressed protein was determined by HPLC/ESI-MS to be 63 770, including the hexahistidine tag. Crystals of recombinant chitinase A were grown to a suitable size for X-ray structure analysis in a precipitant containing 10%(v/v) PEG 400, 0.1 M sodium acetate pH 4.6 and 0.125 M CaCl2. The crystals belonged to the tetragonal space group P422, with two molecules per asymmetric unit and unit-cell parameters a = b = 127.64, c = 171.42 Å. A complete diffraction data set was collected to 2.14 Å resolution using a Rigaku/MSC R-AXIS IV++detector system mounted on an RU-H3R rotating-anode X-ray generator. © 2005 International Union of Crystallography All rights reserved.