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Cell-associated β-xylosidase from Aureobasidium pullulans ATCC 20524: Purification, properties, and characterization of the encoding gene

Paper ID Volume ID Publish Year Pages File Format Full-Text
21499 43225 2010 6 PDF Available
Title
Cell-associated β-xylosidase from Aureobasidium pullulans ATCC 20524: Purification, properties, and characterization of the encoding gene
Abstract

A cell-associated enzyme exhibiting β-xylosidase activity was purified from the cell extract of the dimorphic fungus, Aureobasidium pullulans strain ATCC 20524, grown on oat spelt xylan. The purified enzyme was homogeneous as judged by SDS–PAGE, which showed an apparent Mr of 88.5 kDa. β-Xylosidase activity was optimal at pH 3.5 and 70 °C. The enzyme exhibited apparent Km and Vmax values of 3.5 mM and 263 μmol/mg/min, respectively, for p-nitrophenyl-β-d-xylopyranoside. The enzyme also showed some α-l-arabinofuranosidase activity. Southern blot analysis indicated that the β-xylosidase gene (xylI) was present as a single copy in the genome. The genomic DNA and cDNA encoding this protein were cloned and sequenced. The open reading frame of 2415 bp was interrupted by two introns of 54 and 52 bp, and it encoded a presumed signal peptide of 20 residues and a mature protein of 785 residues with a calculated Mr of 85,045 Da and a deduced isoelectric point of 4.57. The protein was N-glycosylated and possessed 16 potential N-glycosylation sites. Two distinct transcription start points of the xylI gene were present at nt − 32 (A) and − 26 (A) from the start codon. The xylI cDNA was functionally expressed in the yeast Pichia pastoris. The deduced amino acid sequence of the xylI gene product was 49% identical to the Talaromyces emersonii β-xylosidase Bxl1, which belongs to the glycoside hydrolase family 3.

Keywords
DIG, digoxigenin; GH, glycoside hydrolase; nt, nucleotide(s); ORF, open reading frame; pNP, p-nitrophenyl; RACE, rapid amplification of 5′ and 3′ cDNA ends.Aureobasidium pullulans; Cell-associated enzyme; Glycoside hydrolase family 3; Xylan; β-Xylosidase
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Cell-associated β-xylosidase from Aureobasidium pullulans ATCC 20524: Purification, properties, and characterization of the encoding gene
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 110, Issue 2, August 2010, Pages 152–157
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us