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  • Title: Isolation and characterization of genes encoding thermoactive and thermostable dextranases from two thermotolerant soil bacteria.
    Author: Finnegan PM, Brumbley SM, O'Shea MG, Nevalainen H, Bergquist PL.
    Journal: Curr Microbiol; 2004 Nov; 49(5):327-33. PubMed ID: 15486706.
    Abstract:
    Thermotolerant Paenibacillus strain Dex70-1B and unidentified strain Dex70-34 produce thermoactive dextran-degrading enzymes. Plasmid-based genomic DNA libraries constructed from mixed bacterial cultures containing Dex70-1B or Dex70-34 were screened for the ability to confer dextranolytic activity at 70 degrees C onto Escherichia coli. One gene, designated dex1, was isolated from each strain. The Dex70-1B and Dex70-34 dex1 gene sequences were non-identical, and encoded proteins containing 597 (M(r) 68.6 kDa) and 600 amino acids (M(r) 69.2 kDa), respectively. The Dex1 amino acid sequences were most similar to one another, and formed a new clade among the family 66 glycosyl hydrolase sequences. Expression of the Dex1 proteins in E. coli produced dextranolytic activity that converted ethanol-insoluble blue dextran into an ethanol-soluble form, suggestive of endodextranases (EC 3.2.1.11). Both enzymes were most active at about 60 degrees C and pH 5.5, and retained more than 70% maximal activity after incubation at 57 degrees C for 9.5 h in the absence of substrate.
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