2,4-Dichlorophenol Enzymatic Removal and Its Kinetic Study Using Horseradish Peroxidase Crosslinked to Nano Spray-Dried Poly(Lactic-Co-Glycolic Acid) Fine Particles

J Microbiol Biotechnol. 2017 Apr 28;27(4):768-774. doi: 10.4014/jmb.1606.06002.


Horseradish peroxidase (HRP) catalyzes the oxidation of aromatic compounds by hydrogen peroxide via insoluble polymer formation, which can be precipitated from the wastewater. For HRP immobilization, poly(lactic-co-glycolic acid) (PLGA) fine carrier supports were produced by using the Nano Spray Dryer B-90. Immobilized HRP was used to remove the persistent 2,4-dichlorophenol from model wastewater. Both extracted (9-16 U/g) and purified HRP (11-25 U/g) retained their activity to a high extent after crosslinking to the PLGA particles. The immobilized enzyme activity was substantially higher in both the acidic and the alkaline pH regions compared with the free enzyme. Optimally, 98% of the 2,4-dichlorophenol could be eliminated using immobilized HRP due to catalytic removal and partly to adsorption on the carrier supports. Immobilized enzyme kinetics for 2,4-dichlorophenol elimination was studied for the first time, and it could be concluded that competitive product inhibition took place.

Keywords: 2,4-dichlorophenol substrate; Horseradish peroxidase; enzyme kinetics; immobilization.

MeSH terms

  • Adsorption
  • Armoracia / enzymology
  • Catalysis
  • Chlorophenols / metabolism*
  • Enzyme Assays
  • Enzyme Stability
  • Enzymes, Immobilized / metabolism*
  • Horseradish Peroxidase / chemistry
  • Horseradish Peroxidase / metabolism*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Lactic Acid / chemistry*
  • Microscopy, Electron, Scanning
  • Nanoparticles / chemistry*
  • Nanoparticles / ultrastructure
  • Plant Extracts / chemistry
  • Polyglycolic Acid / chemistry*
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Temperature
  • Wastewater
  • Water Purification


  • Chlorophenols
  • Enzymes, Immobilized
  • Plant Extracts
  • Waste Water
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Polyglycolic Acid
  • Lactic Acid
  • Horseradish Peroxidase
  • 2,4-dichlorophenol