Production and identification of extracellular oxidases of Phanerochaete chrysosporium

Leisola, Matti; Meußdoerffer, Franz; Waldner, Roland; Fiechter, Armin

doi:10.1016/0168-1656(85)90024-0

Cited by 68 publications

(60 citation statements)

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“…In Phanerochaete chrysosporium, at least six isoenzymes have been identified (Leisola e t al., 1987). Amino-terminal sequence analyses show that they are encoded by at least four genes (Leisola et al, 1987;Pease & Tien, 1992), two of which have been cloned and characterized (Pease e t al., 1989;Pribnow e t al., 1989). These isoenzymes are differentially regulated in respoc se to nutrient limitation and culture age (Pease & Tien, 1992).…”

Section: Lobos a N D O T H E R Smentioning

confidence: 99%

Isoenzymes of manganese-dependent peroxidase and laccase produced by the lignin-degrading basidiomycete Ceriporiopsis subvermispora

Lobos¹,

Larraı́n²,

Salas³

et al. 1994

Microbiology

141

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The w hi te-ro t basidi om ycete Ceriporiopsis sub vermispora produces two families of ligninolytic enzymes, namely manganese-dependent peroxidases (MnPs) and laccases, when growing in liquid cultures of defined composition. In medium containing 11 p.p.m. of Mn(lI), up to seven isoenzymes of MnP and four isoenzymes of laccase were resolved by isoelectrofocusing (IEF), with pl values in the range 4 4 0 4 6 0 and 3.45-3.65, respectively. Occasionally, a fifth laccase isoform of p l 4 7 0 was also detected. In cultures with 25 and 40 p.p.m. of Mn(ll), mainly the MnPs with higher pl values are produced. The isoenzyme pattern of MnP is not altered throughout the growth period of the fungus. MnP and laccase are also produced by C. subvermispora when growing on wood chips of Pinus radiata. Highest levels of both enzymes were obtained during the first week of incubation. A second peak of MnP activity was observed during the fourth week, whereas very low levels of laccase were extracted from the chips after the second week of growth, IEF analysis showed that the pl values of these laccases are similar to those of laccases produced in liquid cultures, being in the range 3-45-3065. In contrast, four isoforms of MnP were resolved during the first week of incubation on wood chips, with pl values of 440,4*17,4*04 and 3.53. This profile underwent a transition during the second week of growth, a t the end of which isoforms of MnP with pl values of 3*53,340,3-30 and 3.20 were resolved b y IEF. lmmunoblotting studies showed that the molecular mass of MnP isoenzymes from liquid cultures was about 52.5 kDa, whereas the molecular masses of MnPs extracted from wood varied from 52-5 kDa to 62.5 kDa upon ageing of the cultures. The amino terminal sequences of seven MnP isoenzymes were determined. The consensus sequences of MnPs from liquid and solid cultures were clearly distinct, although both showed homology to MnPs from related white-rot fungi.

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Section: Lobos a N D O T H E R Smentioning

confidence: 99%

Isoenzymes of manganese-dependent peroxidase and laccase produced by the lignin-degrading basidiomycete Ceriporiopsis subvermispora

Lobos¹,

Larraı́n²,

Salas³

et al. 1994

Microbiology

141

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“…Other factors, such as 02 partial pressure, agitation, metal ion balance, and pH, are also important (2,19,25,30). Under ligninolytic conditions, P. chrysosporium produces extracellular H202 (10) and a number of H202-requiring peroxidases (ligninases) that oxidize and partially depolymerize lignin or lignin model compounds (14,31,39,41,42).Several schemes have been proposed for the production of extracellular H202 in ligninolytic cultures. These involve either intracellular enzymes, including glucose-l-oxidase (20), glucose-2-oxidase (9), and fatty acyl-coenzyme A oxidase (16), or extracellular Mn2+-dependent peroxidases that reduce 02 to H202 by using NADH, NADPH, or glutathione as an electron source (1,13,34,35).…”

mentioning

confidence: 99%

Involvement of a new enzyme, glyoxal oxidase, in extracellular H2O2 production by Phanerochaete chrysosporium

Kersten¹,

Kirk²

1987

J Bacteriol

337

213

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The importance of extracellular H202 in lignin degradation has become increasingly apparent with the recent discovery of H202-requiring ligninases produced by white-rot fungi. Here we describe a new H202-producing activity of Phanerochaete chrysosporium that involves extraceliular oxidases able to use simple aldehyde, a-hydroxycarbonyl, or a-dicarbonyl compounds as substrates. The activity is expressed during secondary metabolism, when the ligninases are also expressed. Analytical isoelectric focusing of the extracellular proteins, followed by activity staining, indicated that minor proteins with broad substrate specificities are responsible for the oxidase activity. Two of the oxidase substrates, glyoxal and methylglyoxal, were also identified, as their quinoxaline derivatives, in the culture fluid as secondary metabolites. The significance of these findings is discussed with respect to lignin degradation and other proposed systems for H202 production in P. chrysosporium.The white-rot wood-metabolizing basidiomycetes appear to be major degraders of lignin, and, of these, Phanerochaete chrysosporium is the most widely studied species. The ligninolytic system of this organism is expressed in defined media in response to nutrient carbon, nitrogen, or sulfur limitation (19,23,25,38). Other factors, such as 02 partial pressure, agitation, metal ion balance, and pH, are also important (2,19,25,30). Under ligninolytic conditions, P. chrysosporium produces extracellular H202 (10) and a number of H202-requiring peroxidases (ligninases) that oxidize and partially depolymerize lignin or lignin model compounds (14,31,39,41,42).Several schemes have been proposed for the production of extracellular H202 in ligninolytic cultures. These involve either intracellular enzymes, including glucose-l-oxidase (20), glucose-2-oxidase (9), and fatty acyl-coenzyme A oxidase (16), or extracellular Mn2+-dependent peroxidases that reduce 02 to H202 by using NADH, NADPH, or glutathione as an electron source (1,13,34,35).Here we describe a new H202-producing activity, which we term glyoxal oxidase, from P. chrysosporium. Importantly, this activity, as well as two of its many substrates, glyoxal and methylglyoxal, are found in the extracellular fluid of ligninolytic cultures. Extracellular production of H202 seems particularly relevant to the lignin biodegradation process because the ligninases are extracellular and the organism has strong intracellular catalase activity (15). MATERIALS AND METHODSOrganism and culture conditions. P. chrysosporium strain BKM-F-1767 (= ATCC 24725) was grown at 39°C in shallow stationary cultures as previously described (24) Enzyme assays. H202-producing activity was determined by using a modified peroxidase-coupled assay with phenol red as the peroxidase substrate (36). The reaction mixture contained 50 mM Na+ 2,2-dimethylsuccinate (pH 6.0), 10 mM oxidase substrate, 0.01% phenol red, 10 ,ug of horseradish peroxidase (type II; Sigma Chemical Co., St. Louis, Mo.), and up to 300 RI of culture fluid in a total react...

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“…Isozymes differ in physical characteristics, substrate specificity, and stability (8,12), but how these differences relate to lignin degradation is unknown. Production of LiP is derepressed under carbon and nitrogen limitation (17), and the pattern of isozymes formed is influenced by growth conditions (21).…”

mentioning

confidence: 99%

Lignin peroxidase gene family of Phanerochaete chrysosporium: complex regulation by carbon and nitrogen limitation and identification of a second dimorphic chromosome

et al. 1992

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Lignin peroxidases (LiP) ofPhanerochaete chrysosporium are encoded by a family of six closely related genes.Five LiP genes have been localized to the same dimorphic chromosome. In this investigation, relative transcript levels of the LiP genes were determined. Transcripts of the LiPA, LiPB, and 0282 genes were at similar levels in both carbon-and nitrogen-limited cultures. In contrast, transcription of the GLG5, V4, and GLG4 genes was dramatically altered by culture conditions. Under carbon-limited conditions, GLG4 transcripts were, by far, the most abundant. Southern blot analyses of clamped homogeneous field gels were used to map the GLG4 gene to a dimorphic chromosome separate from the other LiP genes.

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Production and identification of extracellular oxidases of Phanerochaete chrysosporium

Cited by 68 publications

References 10 publications

Isoenzymes of manganese-dependent peroxidase and laccase produced by the lignin-degrading basidiomycete Ceriporiopsis subvermispora

Isoenzymes of manganese-dependent peroxidase and laccase produced by the lignin-degrading basidiomycete Ceriporiopsis subvermispora

Involvement of a new enzyme, glyoxal oxidase, in extracellular H2O2 production by Phanerochaete chrysosporium

Lignin peroxidase gene family of Phanerochaete chrysosporium: complex regulation by carbon and nitrogen limitation and identification of a second dimorphic chromosome

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