A novel homodimer laccase from Cerrena unicolor BBP6: Purification, characterization, and potential in dye decolorization and denim bleaching

PLoS One. 2018 Aug 23;13(8):e0202440. doi: 10.1371/journal.pone.0202440. eCollection 2018.

Abstract

The white-rot fungus Cerrena unicolor BBP6 produced up to 243.4 U mL-1 laccase. A novel laccase isoform LacA was purified; LacA is a homodimer with an apparent molecular mass of 55 kDa and an isoelectric point of 4.7. Its optimal pH was 2.5, 4.0, and 5.5 when 2, 2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS), guaiacol, and 2, 6-dimethoxyphenol (2, 6-DMP) were used as the substrates, respectively. The optimal temperature was 60°C for ABTS and 80°C for both guaiacol and 2, 6-DMP. LacA retained 82-92% activity when pH was greater than 4 and 42%-92% activity at or below 50°C. LacA was completely inhibited by 0.1 mM L-cysteine, 1 mM Dithiothreitol, and 10 mM metal ions, Ca2+, Mg2+ and Co2+. LacA had good affinity for ABTS, with a Km of 49.1 μM and a kcat of 3078.9 s-1. It decolorized synthetic dyes at 32.3-87.1%. In the presence of 1-hydroxybenzotriazole (HBT), LacA decolorized recalcitrant dyes such as Safranine (97.1%), Methylene Blue (98.9%), Azure Blue (96.6%) and simulated textile effluent (84.6%). With supplemented manganese peroxidase (MnP), Mn2+ and HBT, the purified LacA and BBP6 fermentation broth showed great potential in denim bleaching, with an up to 5-fold increase in reflectance values.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Coloring Agents / chemistry*
  • Fungal Proteins* / chemistry
  • Fungal Proteins* / isolation & purification
  • Isoenzymes / chemistry
  • Isoenzymes / isolation & purification
  • Laccase* / chemistry
  • Laccase* / isolation & purification
  • Polyporales / enzymology*
  • Textiles*

Substances

  • Coloring Agents
  • Fungal Proteins
  • Isoenzymes
  • Laccase

Grants and funding

This work was fully supported by Ministry of Education, Singapore [MOE2013-TIF-1-G-062, 2014] (AG). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.