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Applied and Environmental Microbiology, December 1999, p. 5546-5553, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The bglA Gene of Aspergillus kawachii Encodes Both Extracellular and Cell Wall-Bound beta -Glucosidases

Kazuhiro Iwashita,* Tatsuya Nagahara, Hitoshi Kimura, Makoto Takano, Hitoshi Shimoi, and Kiyoshi Ito

National Research Institute of Brewing, 7-3-1, Kagamiyama, Higashihiroshima, Hiroshima 739-0046, Japan

Received 26 April 1999/Accepted 27 August 1999

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.

* Corresponding author. Mailing address: National Research Institute of Brewing, 7-3-1, Kagamiyama, Higashihiroshima, Hiroshima 739-0046, Japan. Phone: 81-824-20-0800. Fax: 81-824-20-0809. E-mail: iwasita{at}nrib.go.jp.

Applied and Environmental Microbiology, December 1999, p. 5546-5553, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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