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  • 1
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract A cDNA from Penicillium minioluteum HI-4 encoding a dextranase (1,6-α-glucan hydrolase, EC 3.2.1.11) was isolated and characterized. cDNA clones corresponding to genes expressed in dextran-induced cultures were identified by differential hybridization. Southern hybridization and restriction mapping analysis of selected clones revealed four different groups of cDNAs. The dextranase cDNA was identified after expressing a cDNA fragment from each of the isolated groups of cDNA clones in the Escherichia coli T7 system. The expression of a 2 kb cDNA fragment in E. coli led to the production of a 67 kDa protein which was recognized by an anti-dextranase polyclonal antibody. The cDNA contains 2109 bp plus a poly(A) tail, coding for a protein of 608 amino acids, including 20 N-terminal amino acid residues which might correspond to a signal peptide. There was 29% sequence identity between the P. minioluteum dextranase and the dextranase from Arthrobacter sp. CB-8.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 37 (2000), S. 396-402 
    ISSN: 1432-0983
    Keywords: Key words Dextranase ; Transcriptional regulation ; Induction ; Carbon catabolite repression ; Penicillium minioluteum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The regulation of dextranase (dexA) gene expression in the filamentous fungus Penicillium minioluteum grown on different carbon sources was studied. Growth in the presence of dextran leads to high expression of the dextranase enzyme, but growth in starch, glucose, glycerol, lactose and sorbitol did not. Dextran induced dexA gene expression at the mRNA level. However, in cultures containing dextran plus glucose or glycerol, the transcript was detected 24 h later than in the case where dextran was the only carbon source. When the glucose or glycerol concentration in the dextran-containing medium was kept at about 1% (w/v), no dextranase-transcripts were detected. It was found that both glucose and glycerol inhibited enzyme synthesis, because 1% (w/v) addition of both carbon sources to dextran-growing cultures was able to abolish the inducing effect of dextran. Our results suggest that dextran utilization responds to both specific induction and to glucose and glycerol repression, providing evidence that P. minioluteum dexA expression is regulated by the carbon source at the transcriptional level.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0749-503X
    Keywords: dextranase ; Penicillium minioluteum ; Pichia pastoris ; heterologous gene expression ; protein secretion ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: 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 3·2g/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.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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