High-level production of a thermoacidophilic beta-glucosidase from Penicillium citrinum YS40-5 by solid-state fermentation with rice bran

Bioresour Technol. 2010 Feb;101(4):1310-7. doi: 10.1016/j.biortech.2009.08.049. Epub 2009 Oct 17.

Abstract

A high yield of beta-glucosidase (EC 3.2.1.21) of 159.1 U/g-solid activity on 4-nitrophenyl beta-d-glucopyranoside (pNPG) was achieved by rice bran-based solid-state fermentation (SSF) of the recently characterized fungus Penicillium citrinum YS40-5. The enzyme was both thermophilic and acidophilic at the optimized temperature and pH of 70 degrees C and 5.0, respectively. Over 95% of the original beta-glucosidase activity was maintained after a prolonged storage at ambient temperature for 4 weeks. The kinetic parameters V(max), K(m) and K(I) were 85.93 U/mg, 1.2 mM and 17.59 mM with pNPG, and 72.49 U/mg, 32.17 mM and 8.29 mM with cellobiose, respectively. The protein band with beta-glucosidase activity was characterized by native PAGE followed by MUG-zymogram analysis, and its identity confirmed by nanoLC-MS/MS. A 3.43-fold synergistic effect by combining this beta-glucosidase with Trichoderma reesei cellulases was observed, indicating this enzyme could potentially be used for improving the efficiency of cellulosic bioconversion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Biotechnology / methods*
  • Carbon / pharmacology
  • Cellulose / metabolism
  • DNA, Ribosomal / genetics
  • Enzyme Stability / drug effects
  • Fermentation / drug effects
  • Fermentation / physiology*
  • Glucosides / metabolism
  • Hydrogen-Ion Concentration / drug effects
  • Hymecromone / analogs & derivatives
  • Hymecromone / metabolism
  • Ions
  • Kinetics
  • Mass Spectrometry
  • Metals / pharmacology
  • Molecular Sequence Data
  • Nitrogen / pharmacology
  • Oryza / metabolism*
  • Penicillium / drug effects
  • Penicillium / enzymology*
  • Penicillium / genetics
  • Phylogeny
  • Temperature*
  • Trichoderma / drug effects
  • Trichoderma / enzymology
  • beta-Glucosidase / biosynthesis*
  • beta-Glucosidase / chemistry

Substances

  • DNA, Ribosomal
  • Glucosides
  • Ions
  • Metals
  • 4-methylumbelliferyl glucoside
  • Hymecromone
  • Carbon
  • Cellulose
  • beta-Glucosidase
  • Nitrogen

Associated data

  • GENBANK/EU918770