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Molecular cloning and transcriptional expression analysis of an intracellular beta-glucosidase, a family 3 glycosyl hydrolase, from the edible straw mushroom, Volvariella volvacea

文献类型： 外文期刊

作者： Ding, Shaojun 1 ; Ge, Wei 2 ; Buswell, John A. 3 ;

作者机构： 1.Nanjing Forestry Univ, Dept Biol Engn, Nanjing 210037, Peoples R China

2.Shanghai Acad Agr Sci, Inst Edible Fungi, Shanghai, Peoples R China

3.Chinese Univ Hong Kong, Dept Biol, Shatin, Hong Kong, Peoples R China

关键词： beta-glucosidase;cellulase gene expression;intracellular;Volvariella volvacea;TRICHODERMA-REESEI QM-9414;CELLULOLYTIC SYSTEM;PHANEROCHAETE-CHRYSOSPORIUM;CELLULASE COMPLEX;ENDOGLUCANASE-I;CELL-WALLS;BGL1 GENE;PURIFICATION;INDUCTION;DEGRADATION

期刊名称：FEMS MICROBIOLOGY LETTERS （ 影响因子：2.742； 五年影响因子：2.856 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： A beta-glucosidase, with a molecular mass of 95 kDa, was isolated from extracts of Volvariella volvacea mycelium grown on crystalline cellulose. Degenerate primers based on the N-terminal sequences of purified beta-glucosidase and two protease-generated peptides were used to generate cDNA fragments encoding a portion of the beta-glucosidase gene (bgl), and rapid amplification of cDNA ends was used to obtain full-length cDNA clones. The cDNA of bgl contained an ORF of 2586 bp coding for 862 amino acids. Alignment of the deduced amino-acid sequence of beta-glucosidase with deduced amino acid sequences of other microbial beta-glucosidases showed the highest overall homology with glycoside hydrolase family 3 beta-glucosidases from fungi. Transcripts of bgl were detected in total RNA extracted from mycelium grown on cellulose and cellobiose, and from mycelium pre-grown for 72 h in basal medium containing 1% (w/v) sorbitol following addition of alpha-lactose, beta-lactose, cellobiose, D- xylose, L-sorbose, beta-gentiobiose, sophorose or D-galactose. Addition of L-sorbose and D-glucosamine to mycelium grown on 1% (w/v) crystalline cellulose greatly increased the level of bgl expression. bgl Was expressed at various stages of the mushroom developmental cycle (substrate colonization to mature fruit body), although the number of bgl transcripts in pinhead and button stages was slightly smaller.

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