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Proc Natl Acad Sci U S A. 1990 Apr; 87(8): 2936–2940.

Glyoxal oxidase of Phanerochaete chrysosporium: its characterization and activation by lignin peroxidase.


Glyoxal oxidase (GLOX) is an extracellular H2O2-generating enzyme produced by ligninolytic cultures of Phanerochaete chrysosporium. The production, purification, and partial characterization of GLOX from agitated cultures are described here. High-oxygen levels are critical for GLOX production as for lignin peroxidase. GLOX purified by anion-exchange chromatography appears homogeneous by NaDod-SO4/PAGE (molecular mass = 68 kDa). However, analysis by isoelectric focusing indicates two major bands (pI 4.7 and 4.9) that stain as glycoproteins as well as for H2O2-producing activity in the presence of methylglyoxal. Purified GLOX shows a marked stimulation in activity when incubated with Cu2+; full activation takes more than 1 hr with 1 mM CuSO4 at pH 6. The steady-state kinetic parameters for the GLOX oxidation of methylglyoxal, glyceraldehyde, dihydroxyacetone, glycolaldehyde, acetaldehyde, glyoxal, glyoxylic acid, and formaldehyde, were determined by using a lignin peroxidase coupled-assay at pH 4.5. Of these substrates, the best is the extracellular metabolite methylglyoxal with a Km of 0.64 mM an apparent rate of catalysis, kcat, of 198 s1 under air-saturated conditions. The Km for oxygen is greater than the concentration of oxygen possible at ambient pressure--i.e., >1.3 mM at 25 degrees C. Importantly, oxygen-uptake experiments show that purified GLOX is inactive unless coupled to the peroxidase reaction. With this coupled reaction, for each mol of methylglyoxal, veratryl alcohol (a lignin peroxidase substrate), and oxygen consumed, 1 mol each of pyruvate and veratraldehyde is produced. The importance of these results is discussed in relation to the physiology of lignin biodegradation and possible extracellular regulatory mechanisms for the control of oxidase and peroxidase activities.

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  • Kirk TK, Farrell RL. Enzymatic "combustion": the microbial degradation of lignin. Annu Rev Microbiol. 1987;41:465–505. [PubMed]
  • Tien M. Properties of ligninase from Phanerochaete chrysosporium and their possible applications. Crit Rev Microbiol. 1987;15(2):141–168. [PubMed]
  • Glenn JK, Morgan MA, Mayfield MB, Kuwahara M, Gold MH. An extracellular H2O2-requiring enzyme preparation involved in lignin biodegradation by the white rot basidiomycete Phanerochaete chrysosporium. Biochem Biophys Res Commun. 1983 Aug 12;114(3):1077–1083. [PubMed]
  • Tien M, Kirk TK. Lignin-Degrading Enzyme from the Hymenomycete Phanerochaete chrysosporium Burds. Science. 1983 Aug 12;221(4611):661–663. [PubMed]
  • Glenn JK, Gold MH. Purification and characterization of an extracellular Mn(II)-dependent peroxidase from the lignin-degrading basidiomycete, Phanerochaete chrysosporium. Arch Biochem Biophys. 1985 Nov 1;242(2):329–341. [PubMed]
  • Greene RV, Gould JM. Fatty acyl-coenzyme A oxidase activity and H2O2 production in Phanerochaete chrysosporium mycelia. Biochem Biophys Res Commun. 1984 Jan 30;118(2):437–443. [PubMed]
  • Greene RV, Gould JM. Substrate-induced H2O2 production in mycelia from the lignin-degrading fungus Phanerochaete chrysosporium. Biochem Biophys Res Commun. 1983 Nov 30;117(1):275–281. [PubMed]
  • Kersten PJ, Kirk TK. Involvement of a new enzyme, glyoxal oxidase, in extracellular H2O2 production by Phanerochaete chrysosporium. J Bacteriol. 1987 May;169(5):2195–2201. [PMC free article] [PubMed]
  • Tien M, Kirk TK. Lignin-degrading enzyme from Phanerochaete chrysosporium: Purification, characterization, and catalytic properties of a unique H(2)O(2)-requiring oxygenase. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2280–2284. [PMC free article] [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • RACKER E. The mechanism of action of glyoxalase. J Biol Chem. 1951 Jun;190(2):685–696. [PubMed]
  • Jäger A, Croan S, Kirk TK. Production of Ligninases and Degradation of Lignin in Agitated Submerged Cultures of Phanerochaete chrysosporium. Appl Environ Microbiol. 1985 Nov;50(5):1274–1278. [PMC free article] [PubMed]
  • Gold MH, Kuwahara M, Chiu AA, Glenn JK. Purification and characterization of an extracellular H2O2-requiring diarylpropane oxygenase from the white rot basidiomycete, Phanerochaete chrysosporium. Arch Biochem Biophys. 1984 Nov 1;234(2):353–362. [PubMed]
  • Bar-Lev SS, Kirk TK. Effects of molecular oxygen on lignin degradation by Phanerochaete chrysosporium. Biochem Biophys Res Commun. 1981 Mar 31;99(2):373–378. [PubMed]
  • Tien M, Kirk TK, Bull C, Fee JA. Steady-state and transient-state kinetic studies on the oxidation of 3,4-dimethoxybenzyl alcohol catalyzed by the ligninase of Phanerocheate chrysosporium Burds. J Biol Chem. 1986 Feb 5;261(4):1687–1693. [PubMed]
  • Haemmerli SD, Leisola MS, Sanglard D, Fiechter A. Oxidation of benzo(a)pyrene by extracellular ligninases of Phanerochaete chrysosporium. Veratryl alcohol and stability of ligninase. J Biol Chem. 1986 May 25;261(15):6900–6903. [PubMed]
  • Whittaker MM, Whittaker JW. The active site of galactose oxidase. J Biol Chem. 1988 May 5;263(13):6074–6080. [PubMed]

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