Journal Article

Purification and Characterization of Wall-bound Exo-l,3-β-D-Glucanase from Barley (<i>Hordeum vulgare</i> L.) Seedlings

Toshihisa Kotake, Naoki Nakagawa, Kazuyoshi Takeda and Naoki Sakurai

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 38, issue 2, pages 194-200
Published in print January 1997 | ISSN: 0032-0781
Published online January 1997 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/oxfordjournals.pcp.a029152
Purification and Characterization of Wall-bound Exo-l,3-β-D-Glucanase from Barley (Hordeum vulgare L.) Seedlings

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A β-D-glucanase activity hydrolyzing 1,3:1,4-β-D-glucan was released from the cell walls of barley by 3M LiCl treatment. It was purified by sequential cation-exchange, gel-filtration and hydrophobic chromatography. The molecular mass of the glucanase was 66 kDa as determined by SDS-polyacrylamide gel electrophoresis. Sequence determination of the first thirty amino acids of the N-terminus revealed a high homology of this enzyme to the Pseudomonas l,4-β-D-glucosidase (56.5%). The purified β-D-glucanase has a pH optimum at 5.0, and hydrolyzes oligosaccharides containing β-D-1,3 or β-D-1,4 linkage. The glucanase showed maximum hydrolytic activity toward laminaritetraose, the rate being about two times that of cellotetraose and about four times that of gentiobiose. Polysaccharides such as lichenan, l,3:l,4-β-D-glucan (from barley), laminarin and pustulan are also hydrolyzed, but not carboxylmethyl-curdlan, carboxymethyl-cellulose, xyloglucan and maltose. The purified β-D-glucanase yielded monomeric glucose from laminarihexaose, and exhibited characteristics of an exo-l,3-β-D-glucanase (EC 3.2.1.58). The activity and biochemical characteristics of this enzyme suggest that it is an exo-l,3-β-D-glucanase involved in the rapid turnover of l,3:l,4-β-D-glucan in barley cell walls during seedling growth.

Keywords: Barley; Cell elongation; Cell walls; Exo-glucanase; l,3:l,4-β-D-glucan; Polysaccharide degradation

Journal Article.  0 words. 

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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