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Cloning and expression of a novel Trichoderma viride laminarinase AI gene (lamAI)
Ist Teil von
Bioscience, biotechnology, and biochemistry, 2004-10, Vol.68 (10), p.2111-2119
Ort / Verlag
Tokyo: Japan Society for Bioscience, Biotechnology, and Agrochemistry
Erscheinungsjahr
2004
Quelle
MEDLINE
Beschreibungen/Notizen
The gene lamAl, which encodes a novel laminarinase Al of Dichodenna viride U-1, was cloned using RT-PCR in conjunction with the rapid amplification of cDNA ends (RACE) technique. The open reading frame consisted of 2,277 bp encoding a protein of 759 amino acid residues, including a 32-residue signal prepropeptide. The protein showed 91% sequence similarity to the putative Trichodern a virgins 5-1,3-glucanase BGN1, but no significant similarity to fungal beta-l,6-glucanases or S1,3-glucanases from other organisms. On 40h Incubation with a solo carbon source, northern analysis revealed that the gene was induced by 0.5% laminaran from Eisenia bicyclis but was not by the same concentration of glucose. The lamAl cDNA was functionally expressed in the methylotrophic yeast Pichia pastoris, resulting In a recombinant enzyme with as high activity against laminaran as native LAMAL Based on these data, the probable existence of endo-beta-1,3:1,6-glucan hydrolases as a subclass of endo-p-l,3-glucanases in some mycoparasitic fungi is suggested.