Improvement of laccase production and its properties by low-energy ion implantation

Bioprocess Biosyst Eng. 2010 Jun;33(5):639-46. doi: 10.1007/s00449-009-0389-7. Epub 2009 Oct 31.

Abstract

Low-energy ion implantation was employed to breed laccase producing strain Paecilomyces sp. WSH-L07 and a mutant S152 that exhibited an activity of more than three times over the wild strain was obtained. The optimum substrate of both the wild and mutant laccases was 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate), and followed by guaiacol with optimal pH at 3.4 and 5.0, respectively, while the mutant laccase exhibited a broader active pH range. The mutant laccase had a higher optimal catalytic temperature (60-65 degrees C) than the wild one (55 degrees C), and the wild laccase deactivated rapidly when temperature increased above 55 degrees C. Furthermore, the mutant laccase was more stable under neutral and alkaline conditions. A thermostability experiment revealed that the mutant laccase was superior to the wild laccase. Both laccases were stable in the presence of metal ions, mildly inhibited by SDS (0.5 mM), EDTA (1 mM) and 1,4-dithiothreitol (0.5 mM), and almost completely inhibited by 0.1 mM NaN(3).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzothiazoles / chemistry
  • Fungal Proteins / biosynthesis*
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics
  • Hot Temperature
  • Hydrogen-Ion Concentration
  • Laccase / biosynthesis*
  • Laccase / chemistry
  • Laccase / genetics
  • Mutation, Missense*
  • Sordariales / enzymology*
  • Sordariales / genetics
  • Substrate Specificity
  • Sulfonic Acids / chemistry

Substances

  • Benzothiazoles
  • Fungal Proteins
  • Sulfonic Acids
  • 2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid
  • Laccase