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来自食半乳聚糖嗜琼胶杆菌(Zobellia galactanivorans)的β-琼胶酶催化模块的表达、纯化及初步X射线衍射分析

Expression, purification and preliminary X-ray diffraction analysis of the catalytic module of a beta-agarase from the flavobacterium Zobellia galactanivorans.

作者信息

Hehemann Jan Hendrik, Michel Gurvan, Barbeyron Tristan, Czjzek Mirjam

机构信息

Université Pierre et Marie Curie-Paris 6, Unité Mixte de Recherche 7139 Marine Plants and Biomolecules, Station Biologique, F-29682 Roscoff CEDEX, Bretagne, France.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Apr 1;66(Pt 4):413-7. doi: 10.1107/S174430911000429X. Epub 2010 Mar 31.

Abstract

Marine bacteria secrete specific glycoside hydrolases such as agarases to access polysaccharides from algal cell walls as a carbon and energy source. In an attempt to identify agarases with variable degradation patterns, a novel family GH16 beta-agarase from the marine bacterium Zobellia galactanivorans was expressed, purified and crystallized. The purified enzyme crystallized in two distinct forms that were grown by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. Hexagonal crystals belonging to space group P3(1)21 diffracted to 2.2 A resolution, whereas orthorhombic crystals belonging to space group P2(1)2(1)2(1) diffracted to 1.5 A resolution.

摘要

海洋细菌分泌特定的糖苷水解酶,如琼脂酶,以获取来自藻类细胞壁的多糖作为碳源和能源。为了鉴定具有不同降解模式的琼脂酶,从海洋细菌嗜半乳糖海杆菌中表达、纯化并结晶了一种新的GH16家族β-琼脂酶。纯化后的酶以两种不同的形式结晶,通过悬滴气相扩散法,以聚乙二醇作为沉淀剂生长得到。属于空间群P3(1)21的六方晶体衍射分辨率为2.2 Å,而属于空间群P2(1)2(1)2(1)的正交晶体衍射分辨率为1.5 Å。

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