Identification, purification, and expression patterns of chitinase from psychrotolerant Pedobacter sp. PR-M6 and antifungal activity in vitro

Microb Pathog. 2017 Jun:107:62-68. doi: 10.1016/j.micpath.2017.03.018. Epub 2017 Mar 19.

Abstract

In this study, a novel psychrotolerant chitinolytic bacterium Pedobacter sp. PR-M6 that displayed strong chitinolytic activity on 0.5% colloidal chitin was isolated from the soil of a decayed mushroom. Chitinase activity of PR-M6 at 25 °C (C25) after 6 days of incubation with colloidal chitin increased rapidly to a maximum level (31.3 U/mg proteins). Three chitinase isozymes (chiII, chiIII, and chiIV) from the crude enzyme at 25 °C (C25) incubation were expressed on SDS-PAGE gels at 25 °C. After purification by chitin-affinity chromatography, six chitinase isozymes (chiI, chiII, chiIII, chiIV, chiV, and chiVI) from C25-fractions were expressed on SDS-PAGE gels at 25 °C. Major bands of chitinase isozymes (chiI, chiII, and chiIII) from C4-fractions were strongly expressed on SDS-PAGE gels at 25 °C. Pedobacter sp. PR-M6 showed high inhibition rate of 60.9% and 57.5% against Rhizoctonia solani and Botrytis cinerea, respectively. These results indicated that psychrotolerant Pedobacter sp. PR-M6 could be applied widely as a microorganism agent for the biocontrol of agricultural phytopathogens at low temperatures.

Keywords: Chitinase isozyme; Chitinolytic activity; Pedobacter sp.; Psychrotolerance.

MeSH terms

  • Agriculture
  • Antifungal Agents / isolation & purification*
  • Biological Control Agents / isolation & purification
  • Botrytis / drug effects
  • Chitin / metabolism
  • Chitinases / antagonists & inhibitors
  • Chitinases / biosynthesis*
  • Chitinases / chemistry*
  • Chitinases / isolation & purification*
  • Chromatography, Affinity / methods
  • Cold Temperature
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Assays
  • Isoenzymes / chemistry
  • Isoenzymes / isolation & purification
  • Mycelium / drug effects
  • Mycelium / growth & development
  • Pedobacter / classification
  • Pedobacter / enzymology*
  • Pedobacter / growth & development
  • Pedobacter / isolation & purification
  • Phylogeny
  • Rhizoctonia / drug effects
  • Soil Microbiology

Substances

  • Antifungal Agents
  • Biological Control Agents
  • Isoenzymes
  • Chitin
  • Chitinases