Archives of Microbiology

, 184:215

Degradation of rice bran hemicellulose by Paenibacillus sp. strain HC1: gene cloning, characterization and function of β-D-glucosidase as an enzyme involved in degradation

  • Karen Mine Harada
  • Keiko Tanaka
  • Yasuki Fukuda
  • Wataru Hashimoto
  • Kousaku Murata
Original Paper

DOI: 10.1007/s00203-005-0038-8

Cite this article as:
Harada, K.M., Tanaka, K., Fukuda, Y. et al. Arch Microbiol (2005) 184: 215. doi:10.1007/s00203-005-0038-8
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Abstract

A bacterium (strain HC1) capable of assimilating rice bran hemicellulose was isolated from a soil and identified as belonging to the genus Paenibacillus through taxonomical and 16S rDNA sequence analysis. Strain HC1 cells grown on rice bran hemicellulose as a sole carbon source inducibly produced extracellular xylanase and intracellular glycosidases such as β-d-glucosidase and β-d-arabinosidase. One of them, β-d-glucosidase was further analyzed. A genomic DNA library of the bacterium was constructed in Escherichia coli and gene coding for β-d-glucosidase was cloned by screening for β-d-glucoside-degrading phenotype in E. coli cells. Nucleotide sequence determination indicated that the gene for the enzyme contained an open reading frame consisting of 1,347 bp coding for a polypeptide with a molecular mass of 51.4 kDa. The polypeptide exhibits significant homology with other bacterial β-d-glucosidases and belongs to glycoside hydrolase family 1. β-d-Glucosidase purified from E. coli cells was a monomeric enzyme with a molecular mass of 50 kDa most active at around pH 7.0 and 37°C. Strain HC1 glycosidases responsible for degradation of rice bran hemicellulose are expected to be useful for structurally determining and molecularly modifying rice bran hemicellulose and its derivatives.

Keywords

Rice bran Hemicellulose Degradation Paenibacillus β-d-glucosidase 

Abbreviations

RB

Rice bran

TLC

Thin-layer chromatography

X-Glc

5-Bromo-4-chrolo-3-indoryl-β-d-glucopyranoside

p-NP

p-nitrophenyl

HPLC

High-performance liquid chromatography

KPB

Potassium phosphate buffer

SDS-PAGE

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

aa

Amino acids

acc no

Accession number

bglA

Strain HC1 β-d-glucosidase gene

BglA

Strain HC1 β-d-glucosidase

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Karen Mine Harada
    • 1
  • Keiko Tanaka
    • 2
  • Yasuki Fukuda
    • 2
  • Wataru Hashimoto
    • 1
  • Kousaku Murata
    • 1
  1. 1.Laboratory of Basic and Applied Molecular Biotechnology, Division of Food Science and Biotechnology, Graduate School of AgricultureKyoto UniversityUji, KyotoJapan
  2. 2.Chukyo Community CollegeMizunami, GifuJapan

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