来自豌豆镰刀菌的重组角质酶的结晶及初步X射线研究。

Crystallization and preliminary X-ray study of a recombinant cutinase from Fusarium solani pisi.

作者信息

Abergel C, Martinez C, Fontecilla-Camps J, Cambillau C, de Geus P, Lauwereys M

机构信息

Laboratoire de Cristallographie et Cristallisation des Macromolécules Biologiques URA-1296 CNRS, Faculté de Médecine Nord, Marseille, France.

出版信息

J Mol Biol. 1990 Sep 20;215(2):215-6. doi: 10.1016/S0022-2836(05)80339-0.

DOI:10.1016/S0022-2836(05)80339-0

PMID:2213880

Abstract

Recombinant cutinase from Fusarium solani pisi is expressed and excreted with very high yields in Escherichia coli cultures. Cutinase was crystallized at 20 degrees C using the vapour diffusion technique, with polyethylene glycol 6000 as precipitant. Best crystals were obtained at pH 7.0 with polyethylene glycol 6000 as precipitant. Best crystals were obtained at pH 7.0 with polyethylene glycol at 15 to 20%. They are monoclinic, with space group P2(1) and cell dimensions a = 35.1 A, b = 67.4 A, c = 37.05 A and beta = 94.0 degrees; they diffract beyond 1.5 A resolution. The asymmetric unit contains one molecule of 22,000 Da (Vm = 1.98 A3/Da; 38% water).

摘要

来自腐皮镰刀菌的重组角质酶在大肠杆菌培养物中能够以非常高的产量表达并分泌。利用蒸汽扩散技术，以聚乙二醇6000作为沉淀剂，在20℃下使角质酶结晶。在pH 7.0、聚乙二醇浓度为15%至20%时获得了最佳晶体。它们是单斜晶系，空间群为P2(1)，晶胞参数a = 35.1 Å，b = 67.4 Å，c = 37.05 Å，β = 94.0°；其衍射分辨率超过1.5 Å。不对称单元包含一个22,000 Da的分子（Vm = 1.98 Å3/Da；38%为水）。