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  • Title: Cloning of the Penicillium minioluteum gene encoding dextranase and its expression in Pichia pastoris.
    Author: Roca H, Garcia B, Rodriguez E, Mateu D, Coroas L, Cremata J, Garcia R, Pons T, Delgado J.
    Journal: Yeast; 1996 Sep 30; 12(12):1187-200. PubMed ID: 8905923.
    Abstract:
    The DEX gene encoding an extracellular dextranase was isolated from the genomic DNA library of Penicillium minioluteum by hybridization using the dextranase cDNA as a probe. Comparison of the gene and cDNA sequences revealed that the DEX gene does not contain introns. Amino acid sequences comparison of P. minioluteum dextranase with other reported dextranases reveals a significant homology (29% identity) with a dextranase from Arthrobacter sp. CB-8. The DEX gene fragment encoding a mature protein of 574 amino acids was expressed in the methylotrophic yeast Pichia pastoris by using the SUC2 gene signal sequence from Saccharomyces cerevisiae under control of the alcohol oxidase-1 (AOX1) promoter. Over 3.2 g/l of enzymatically active dextranase was secreted into the medium after induction by methanol. The yeast product was indistinguishable from the native enzyme in specific activity and the N-terminus of both proteins were identical.
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