Crystallization and preliminary crystallographic analysis of beta-L-arabinopyranosidase from Streptomyces avermitilis NBRC14893

Abstract

Beta-L-arabinopyranosidase from Streptomyces avermitilis NBRC14893 is a monomeric protein consisting of a catalytic domain belonging to glycosyl hydrolase family 27, an unknown domain and a substrate-binding domain belonging to carbohydrate-binding module family 13. The complete enzyme (residues 45-658) has successfully been cloned and homologously expressed in the Streptomyces expression system. beta-L-Arabinopyranosidase was crystallized by the sitting-drop vapour-diffusion method. The crystals diffracted to 1.6 A resolution and belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 68.2, b = 98.9, c = 181.3 A. The Matthews coefficient was calculated to be 2.38 A(3) Da(-1).

Figure 1

SDS–PAGE of recombinant β-l-arabinopyranosidase. The protein was subjected to SDS–PAGE on a 12% polyacrylamide gel and the bands were visualized by Coomassie Brilliant Blue R-250 staining. Lane 1, molecular-weight markers (1 µg each); lane 2, purified recombinant β-l-arabinopyranosidase (1 µg). Molecular weights are given in kDa at the side of the gel.

Figure 2

Crystals of β-l-arabinopyranosidase. The scale bar represents 500 µm.