Heterologous expression of cDNAs encoding barley (Hordeum vulgare) (1-->3)-beta-glucanase isoenzyme GV

Abstract

Two cDNAs have been isolated from libraries generated from poly(A)+RNA of young barley roots and leaves, using a cDNA encoding barley (1-->3)-beta-glucanase isoenzyme GII as a probe. Nucleotide sequence analyses and ribonuclease protection assays show that the two cDNAs differ only in the length of their 3'-untranslated regions; the corresponding mRNAs are likely to originate from a single gene by tissue-specific processing at separate polyadenylation sites. When the coding region of the cDNA is expressed in E. coli, the resultant protein catalyses the hydrolysis of (1-->3)-beta-glucan with an action pattern characteristic of a (1-->3)-beta-glucan endohydrolase (EC 3.2.1.39). The enzyme has been designated isoenzyme GV of the barley (1-->3)-beta-glucanase family).