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Appl Environ Microbiol. 1999 Dec;65(12):5546-53.

The bglA gene of Aspergillus kawachii encodes both extracellular and cell wall-bound beta-glucosidases.

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  • 1National Research Institute of Brewing, 7-3-1, Kagamiyama, Higashihiroshima, Hiroshima 739-0046, Japan. iwasita@nrib.go.jp

Abstract

We cloned the genomic DNA and cDNA of bglA, which encodes beta-glucosidase in Aspergillus kawachii, based on a partial amino acid sequence of purified cell wall-bound beta-glucosidase CB-1. The nucleotide sequence of the cloned bglA gene revealed a 2,933-bp open reading frame with six introns that encodes an 860-amino-acid protein. Based on the deduced amino acid sequence, we concluded that the bglA gene encodes cell wall-bound beta-glucosidase CB-1. The amino acid sequence exhibited high levels of homology with the amino acid sequences of fungal beta-glucosidases classified in subfamily B. We expressed the bglA cDNA in Saccharomyces cerevisiae and detected the recombinant beta-glucosidase in the periplasm fraction of the recombinant yeast. A. kawachii can produce two extracellular beta-glucosidases (EX-1 and EX-2) in addition to the cell wall-bound beta-glucosidase. A. kawachii in which the bglA gene was disrupted produced none of the three beta-glucosidases, as determined by enzyme assays and a Western blot analysis. Thus, we concluded that the bglA gene encodes both extracellular and cell wall-bound beta-glucosidases in A. kawachii.

PMID:
10584016
[PubMed - indexed for MEDLINE]
PMCID:
PMC91756
Free PMC Article
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