Kinetic analysis of manganese peroxidase. The reaction with manganese complexes

I. C. Kuan, K. A. Johnson, M. Tien

Research output: Contribution to journalArticle

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Abstract

Manganese peroxidase from the lignin-degrading fungus Phanerochaete chrysosporium catalyzes the H2O2-dependent oxidation of Mn2+ to Mn3+. Presteady-state methods were employed to characterize the reactions of free and chelated Mn2+ with the 2-electron and 1-electron oxidized forms of the enzyme, compounds I and II, respectively. At pH 4.5, the optimum pH for steady-state turnover, the reaction of compound I with Mn2+, either free or complexed, is too rapid to measure by stopped flow methods. The reactions of compound I with Mn2+ can only be monitored under non-optimal conditions of pH 2.5. The reaction of compound II with Mn2+ is much slower than compound I. Chelators such as oxalate, lactate, and malonate facilitated the reaction of Mn2+ with compound II. In contrast, succinate, which does not readily form a complex with Mn2+, and polyglutamate, which is polymeric, were ineffective in stimulating the reaction of Mn2+ with compound II. The 1:1 chelator-Mn2+ complex is the preferred substrate for compound II; this conclusion is based on known formation constants for the various Mn2+ complexes. Steady-state kinetics studies were performed by directly measuring the initial rate of Mn3+ formation. The k(cat) values for the formation of Mn3+-oxalate, Mn3+-lactate, and Mn3+-malonate are 308, 211, and 220 s- 1, respectively. The K(m) values for Mn2+-oxalate, Mn2+-lactate, and Mn2+-malonate are 13, 41, and 18 μM, respectively. These results collectively indicate that manganese peroxidase does not readily oxidize free (hexa-aquo) Mn2+ as previously proposed (Wariishi, H., Valli, K., and Gold, M. H. (1992) J. Biol. Chem. 267, 23688-23695), but the Mn2+ has to be chelated to support steady-state turnover.

Original languageEnglish (US)
Pages (from-to)20064-20070
Number of pages7
JournalJournal of Biological Chemistry
Volume268
Issue number27
StatePublished - Jan 1 1993

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manganese peroxidase
Oxalates
Manganese
Lactic Acid
Chelating Agents
Kinetics
Electrons
Phanerochaete
Polyglutamic Acid
Lignin
Succinic Acid
Fungi
Oxidation
Substrates
Enzymes
malonic acid

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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title = "Kinetic analysis of manganese peroxidase. The reaction with manganese complexes",
abstract = "Manganese peroxidase from the lignin-degrading fungus Phanerochaete chrysosporium catalyzes the H2O2-dependent oxidation of Mn2+ to Mn3+. Presteady-state methods were employed to characterize the reactions of free and chelated Mn2+ with the 2-electron and 1-electron oxidized forms of the enzyme, compounds I and II, respectively. At pH 4.5, the optimum pH for steady-state turnover, the reaction of compound I with Mn2+, either free or complexed, is too rapid to measure by stopped flow methods. The reactions of compound I with Mn2+ can only be monitored under non-optimal conditions of pH 2.5. The reaction of compound II with Mn2+ is much slower than compound I. Chelators such as oxalate, lactate, and malonate facilitated the reaction of Mn2+ with compound II. In contrast, succinate, which does not readily form a complex with Mn2+, and polyglutamate, which is polymeric, were ineffective in stimulating the reaction of Mn2+ with compound II. The 1:1 chelator-Mn2+ complex is the preferred substrate for compound II; this conclusion is based on known formation constants for the various Mn2+ complexes. Steady-state kinetics studies were performed by directly measuring the initial rate of Mn3+ formation. The k(cat) values for the formation of Mn3+-oxalate, Mn3+-lactate, and Mn3+-malonate are 308, 211, and 220 s- 1, respectively. The K(m) values for Mn2+-oxalate, Mn2+-lactate, and Mn2+-malonate are 13, 41, and 18 μM, respectively. These results collectively indicate that manganese peroxidase does not readily oxidize free (hexa-aquo) Mn2+ as previously proposed (Wariishi, H., Valli, K., and Gold, M. H. (1992) J. Biol. Chem. 267, 23688-23695), but the Mn2+ has to be chelated to support steady-state turnover.",
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Kinetic analysis of manganese peroxidase. The reaction with manganese complexes. / Kuan, I. C.; Johnson, K. A.; Tien, M.

In: Journal of Biological Chemistry, Vol. 268, No. 27, 01.01.1993, p. 20064-20070.

Research output: Contribution to journalArticle

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N2 - Manganese peroxidase from the lignin-degrading fungus Phanerochaete chrysosporium catalyzes the H2O2-dependent oxidation of Mn2+ to Mn3+. Presteady-state methods were employed to characterize the reactions of free and chelated Mn2+ with the 2-electron and 1-electron oxidized forms of the enzyme, compounds I and II, respectively. At pH 4.5, the optimum pH for steady-state turnover, the reaction of compound I with Mn2+, either free or complexed, is too rapid to measure by stopped flow methods. The reactions of compound I with Mn2+ can only be monitored under non-optimal conditions of pH 2.5. The reaction of compound II with Mn2+ is much slower than compound I. Chelators such as oxalate, lactate, and malonate facilitated the reaction of Mn2+ with compound II. In contrast, succinate, which does not readily form a complex with Mn2+, and polyglutamate, which is polymeric, were ineffective in stimulating the reaction of Mn2+ with compound II. The 1:1 chelator-Mn2+ complex is the preferred substrate for compound II; this conclusion is based on known formation constants for the various Mn2+ complexes. Steady-state kinetics studies were performed by directly measuring the initial rate of Mn3+ formation. The k(cat) values for the formation of Mn3+-oxalate, Mn3+-lactate, and Mn3+-malonate are 308, 211, and 220 s- 1, respectively. The K(m) values for Mn2+-oxalate, Mn2+-lactate, and Mn2+-malonate are 13, 41, and 18 μM, respectively. These results collectively indicate that manganese peroxidase does not readily oxidize free (hexa-aquo) Mn2+ as previously proposed (Wariishi, H., Valli, K., and Gold, M. H. (1992) J. Biol. Chem. 267, 23688-23695), but the Mn2+ has to be chelated to support steady-state turnover.

AB - Manganese peroxidase from the lignin-degrading fungus Phanerochaete chrysosporium catalyzes the H2O2-dependent oxidation of Mn2+ to Mn3+. Presteady-state methods were employed to characterize the reactions of free and chelated Mn2+ with the 2-electron and 1-electron oxidized forms of the enzyme, compounds I and II, respectively. At pH 4.5, the optimum pH for steady-state turnover, the reaction of compound I with Mn2+, either free or complexed, is too rapid to measure by stopped flow methods. The reactions of compound I with Mn2+ can only be monitored under non-optimal conditions of pH 2.5. The reaction of compound II with Mn2+ is much slower than compound I. Chelators such as oxalate, lactate, and malonate facilitated the reaction of Mn2+ with compound II. In contrast, succinate, which does not readily form a complex with Mn2+, and polyglutamate, which is polymeric, were ineffective in stimulating the reaction of Mn2+ with compound II. The 1:1 chelator-Mn2+ complex is the preferred substrate for compound II; this conclusion is based on known formation constants for the various Mn2+ complexes. Steady-state kinetics studies were performed by directly measuring the initial rate of Mn3+ formation. The k(cat) values for the formation of Mn3+-oxalate, Mn3+-lactate, and Mn3+-malonate are 308, 211, and 220 s- 1, respectively. The K(m) values for Mn2+-oxalate, Mn2+-lactate, and Mn2+-malonate are 13, 41, and 18 μM, respectively. These results collectively indicate that manganese peroxidase does not readily oxidize free (hexa-aquo) Mn2+ as previously proposed (Wariishi, H., Valli, K., and Gold, M. H. (1992) J. Biol. Chem. 267, 23688-23695), but the Mn2+ has to be chelated to support steady-state turnover.

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