Cloning and Expression of an α-1,3-Glucanase Gene from Bacillus circulans KA-304 : The Enzyme Participates in Protoplast Formation of Schizophyllum commune
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A culture filtrate of <I>Bacillus circulans</I> KA-304 grown on a cell-wall preparation of <I>Schizophyllum commune</I> has an activity to form protoplasts from <I>S. commune</I> mycelia, and a combination of α-1,3-glucanase and chitinase I, which were isolated from the filtrate, brings about the protoplast-forming activity.<BR>The gene of α-1,3-glucanase was cloned from <I>B. circulans</I> KA-304. It consists of 3,879 nucleotides, which encodes 1,293 amino acids including a putative signal peptide (31 amino acid residues), and the molecular weight of α-1,3-glucanase without the putative signal peptide was calculated to be 132,184. The deduced amino acid sequence of α-1,3-glucanase of <I>B. circulans</I> KA-304 showed approximately 80% similarity to that of mutanase (α-1,3-glucanase) of <I>Bacillus sp.</I> RM1, but no significant similarity to those of fungal mutanases.<BR>The recombinant α-1,3-glucanase was expressed in <I>Escherichia coli</I> Rosetta-gami B (DE 3), and significant α-1,3-glucanase activity was detected in the cell-free extract of the organism treated with isopropyl-β-<small>D</small>-thiogalactopyranoside. The recombinant α-1,3-glucanase showed protoplast-forming activity when the enzyme was combined with chitinase I.
- Agricultural and Biological Chemistry
Agricultural and Biological Chemistry 70(7), 1754-1763, 2006-07-23
Japan Society for Bioscience, Biotechnology, and Agrochemistry