Thermostable invertases from Paecylomyces variotii produced under submerged and solid-state fermentation using agroindustrial residues

World J Microbiol Biotechnol. 2012 Feb;28(2):463-72. doi: 10.1007/s11274-011-0837-9. Epub 2011 Jul 12.

Abstract

The filamentous fungus Paecylomices variotii was able to produce high levels of cell extract and extracellular invertases when grown under submerged fermentation (SbmF) and solid-state fermentation, using agroindustrial products or residues as substrates, mainly soy bran and wheat bran, at 40°C for 72 h and 96 h, respectively. Addition of glucose or fructose (≥1%; w/v) in SbmF inhibited enzyme production, while the addition of 1% (w/v) peptone as organic nitrogen source enhanced the production by 3.7-fold. However, 1% (w/v) (NH(4))(2)HPO(4) inhibited enzyme production around 80%. The extracellular form was purified until electrophoretic homogeneity (10.5-fold with 33% recovery) by DEAE-Fractogel and Sephacryl S-200 chromatography. The enzyme is a monomer with molecular mass of 102 kDa estimated by SDS-PAGE with carbohydrate content of 53.6%. Optima of temperature and pH for both, extracellular and cell extract invertases, were 60°C and 4.0-4.5, respectively. Both invertases were stable for 1 h at 60°C with half-lives of 10 min at 70°C. Mg(2+), Ba(2+) and Mn(2+) activated both extracellular and cell extract invertases from P. variotii. The kinetic parameters K(m) and V(max) for the purified extracellular enzyme corresponded to 2.5 mM and 481 U/mg prot(-1), respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Enzyme Stability
  • Fermentation / physiology*
  • Fungal Proteins / chemistry
  • Fungal Proteins / isolation & purification*
  • Fungal Proteins / metabolism*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Sucrose / metabolism
  • Temperature
  • beta-Fructofuranosidase / chemistry
  • beta-Fructofuranosidase / isolation & purification*
  • beta-Fructofuranosidase / metabolism*

Substances

  • Fungal Proteins
  • Sucrose
  • beta-Fructofuranosidase