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番木瓜乳胶中19家族糖基水解酶的结晶及初步X射线分析

Crystallization and preliminary X-ray analysis of a family 19 glycosyl hydrolase from Carica papaya latex.

作者信息

Huet Joëlle, Azarkan Mohamed, Looze Yvan, Villeret Vincent, Wintjens René

机构信息

Laboratoire de Chimie Générale (CP 206/4), Institut de Pharmacie, Université Libre de Bruxelles (ULB), Campus de la Plaine, Boulevard du Triomphe, B-1050 Bruxelles, Belgium.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 May 1;64(Pt 5):371-4. doi: 10.1107/S1744309108007823. Epub 2008 Apr 5.

Abstract

A chitinase isolated from the latex of the tropical species Carica papaya has been purified to homogeneity and crystallized. This enzyme belongs to glycosyl hydrolase family 19 and exhibits exceptional resistance to proteolysis. The initially observed crystals, which diffracted to a resolution of 2.0 A, were improved through modification of the crystallization protocol. Well ordered crystals were subsequently obtained using N-acetyl-D-glucosamine, the monomer resulting from the hydrolysis of chitin, as an additive to the crystallization solution. Here, the characterization of a chitinase crystal that belongs to the monoclinic space group P2(1), with unit-cell parameters a = 69.08, b = 44.79, c = 76.73 A, beta = 95.33 degrees and two molecules per asymmetric unit, is reported. Diffraction data were collected to a resolution of 1.8 A. Structure refinement is currently in progress.

摘要

从热带物种番木瓜的乳胶中分离出的一种几丁质酶已被纯化至同质并结晶。这种酶属于糖基水解酶家族19,对蛋白水解表现出非凡的抗性。最初观察到的晶体衍射分辨率为2.0埃,通过改进结晶方案得到了改善。随后,使用几丁质水解产生的单体N-乙酰-D-葡萄糖胺作为结晶溶液的添加剂,获得了排列良好的晶体。在此,报道了一种属于单斜空间群P2(1)的几丁质酶晶体的表征,其晶胞参数为a = 69.08,b = 44.79,c = 76.73埃,β = 95.33°,每个不对称单元有两个分子。衍射数据收集到1.8埃的分辨率。结构精修目前正在进行中。

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