학술논문
Expression, purification, crystallization and preliminary crystallographic analysis of chitinase A from Vibrio carchariae.
Document Type
Article
Author
Source
Subject
*ESCHERICHIA coli
*CRYSTALS
*PROTEINS
*CHITINASE
*SODIUM acetate
*X-ray diffraction
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Language
ISSN
1744-3091
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 Awere 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 unitcell 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. [ABSTRACT FROM AUTHOR]