Purification and characterization of prophenoloxidase from Galleria mellonella L

Artif Cells Blood Substit Immobil Biotechnol. 2012 Dec;40(6):391-5. doi: 10.3109/10731199.2012.696060. Epub 2012 Jul 10.

Abstract

Prophenoloxidase (PPO) was purified from Galleria mellonella L. A 67-fold purification of the proenzyme with 352% yield was achieved by using a Sepharose 4B-L-tyrosine-p-amino benzoic acid affinity column. The purified enzyme was migrated as a single band on SDS-polyacrylamide gel electrophoresis. K(m) and V(max) values were 0.017 M and 1430.45 EU for catechol. Inhibition of PPO was investigated with inhibitors such as p-aminobenzoic acid, etyleneglycol, and ascorbic acid. Among them, ascorbic acid showed the strongest inhibitory activity with IC(50) value of 2.94 μM. The current paper represents new strategies for the biological control of the Galleria mellonella L. insect.

MeSH terms

  • 4-Aminobenzoic Acid / chemistry
  • Animals
  • Catechol Oxidase / chemistry*
  • Catechol Oxidase / isolation & purification
  • Chromatography, Agarose / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Inhibitors / chemistry*
  • Enzyme Precursors / chemistry*
  • Enzyme Precursors / isolation & purification
  • Hemolymph / enzymology*
  • Hydrogen-Ion Concentration
  • Insect Control / methods
  • Larva / enzymology
  • Moths / enzymology*
  • Substrate Specificity
  • Temperature
  • Tyrosine / chemistry

Substances

  • Enzyme Inhibitors
  • Enzyme Precursors
  • Tyrosine
  • pro-phenoloxidase
  • Catechol Oxidase
  • 4-Aminobenzoic Acid