Etienne Odier

Effect of Tween 80 and oleic acid on ligninase production by Phanerochaete chrysosporium INA-12

Ligninase production by Phanerochaete chrysosporium INA-12 was increased when culture medium was supplemented with sorbitan polyoxyethylene monooleate (Tween 80), oleic acid alone or emulsified with Tween 80. Results showed that fatty acids contained in Tween 80 were dissimilated by P. chrysosporium INA-12 in glycerol-containing cultures. Lipase activity (EC 3.1.1.3) was detected in the culture filtrate after one day of incubation. Ligninase production was markedly enhanced and fermentation time for maximum activity was reduced in presence of exogenous oleic acid emulsified with Tween 80 (0.04%, w/v). Ligninase activity in stationary and agitated conditions were 22.4 and 19.3 nkat ml−1respectively. Agitated cultures containing Tween 80 or emulsified oleic acid converted [ring-U-14C]lignin to14CO2at a rate in proportion to ligninase activity. The effect of additives as well as the development of suitable culture conditions for ligninase production by P. chrysosporium INA-12 using submerged agitated cultures is discussed.

Phospholipid and fatty acid enrichment of Phanerochaete chrysosporium INA-12 in relation to ligninase production

SummaryLigninase activity of Phanerochaete chrysosporium INA-12 was increased when vegetable oils emulsified with sorbitan polyoxyethylene monooleate (Tween 80) were added to growth medium. Maximal enzyme yield was 22.0 nkat·ml-1 in olive oil cultures after 4 days incubation. P. chrysosporium INA-12 was also able to utilize tall oil fatty acids for ligninase synthesis. An extracellular lipase activity was detected during the primary phase of growth in culture containing vegetable oils. On the other hand, ligninase production was 1.5-fold enhanced when olive oil cultures were supplemented with soybean asolectin as a phospholipid source. In cultures supplied with olive oil plus asolectin, P. chrysosporium INA-12 mycelium exhibited a preferential enrichment of oleic acid (C18:1), phosphatidylcholine (PC) and lysophosphatidylcholine (LPC) as compared to lipid-free medium. PC and LPC enrichment was associated with an increased ratio of saturated versus unsaturated fatty acids of phospholipids.

Ligninase-mediated phenoxy radical formation and polymerization unaffected by cellobiose: quinone oxidoreductase☆

Phanerochete chrysosporium ligninase (+ H2O2) oxidized the lignin substructure-related compound acetosyringone to a phenoxy radical which was identified by ESR spectroscopy. Cellobiose:quinone oxidoreductase (CBQase) + cellobiose, previously suggested to be a phenoxy radical reducing system, was without effect on the radical. Ligninase polymerized guaiacol and it increased the molecular size of a synthetic lignin. These polymerizations, reflecting phenoxy radical coupling reactions, were also unaffected by the CBQase system. We conclude that ligninase catalyzes phenol polymerization via phenoxy radicals, which CBQase does not affect. The CBQase system also did not produce H2O2, and its physiological role remains obscure. Glucose oxidase + glucose did produce H2O2 as expected, but, like CBQase, it did not reduce the phenoxy radical of acetosyringone. Because intact cultures of P. chrysosporium depolymerize lignins, it is likely that phenol polymerization by ligninase is prevented or reversed in vivo by an as yet undescribed system.

Influence of the physical state of lignin on its degradability by the lignin peroxidase of Phanerochaete chrysosporium

Abstract The rate of oxidation of spruce milled wood lignin by Phanerochaete chrysosporium was investigated with precipitated lignin as well as lignin finely dispersed in water (colloidal lignin). H2O2 consumption rates in the course of lignin oxidation in the presence of purified lignin peroxidase were much higher with colloidal lignin compared with precipitated lignin. Characterization of reaction products and molecular-size distribution confirms that reaction rate of lignin peroxidase with 14C-lignin is strongly dependent on the physical state of lignin. The lignin peroxidase catalyse mainly repolymerization of colloidal lignin. However, in the presence of veratryl alcohol, both polymerization and depolymerization were observed.

Lignin peroxidase production by strains of Phanerochaete chrysosporium grown on glycerol

SummaryLignin peroxidase production by several strains of Phanerochaete chrysosporium was determined during growth on glycerol under conditions of nitrogen sufficiency. Fungal strains which grew poorest on glycerol produced the highest titres of lignin peroxidase whereas enzyme levels were much lower when marginally greater biomass values were recorded. In the case of P. chrysosporium strain INA-12, the nature of the nitrogen source had a pronounced effect on both growth and enzyme production. Highest biomass values were obtained when l-glutamate or l-glutamine served as the major nitrogen source but enzyme synthesis was normally repressed completely. Lignin peroxidase activity in this strain was maximal when the initial pH of the culture medium was adjusted to pH 5.0.

Nitrogen and carbon regulation of lignin peroxidase and enzymes of nitrogen metabolism inPhanerochaete chrysosporium

The regulation of nitrogen metabolism pathways was examined inPhanerochaete chrysosporium in relation to the repression of lignin peroxidase by nitrogen or carbon in this fungus. Under conditions of nitrogen derepression,P. chrysosporium synthesizes the amidohydrolases, formamidase (EC 3.5.1.9) and acetamidase (EC 3.5.1.4) and the enzymes of purine catabolism uricase (EC 1.7.3.3), allantoinase (EC 3.5.2.5), and allantoicase (EC 3.5.3.4). Formamidase is repressed to low levels in the presence of ammonium and there is no apparent control of this enzyme by carbon catabolite repression. Although formamide is a nitrogen source, it is not a carbon source forP. chrysosporium. Glutamate totally represses formamidase. Uricase, allantoinase, and allantoicase are also regulated by nitrogen repression but not carbon catabolite repression. Urease is synthesized at similar levels irrespective of the nitrogen or carbon conditions. The sensitivity of uricase, allantoinase, and allantoicase to nitrogen repression is less than that of formamidase. In contrast to formamidase, glutamate is not a more powerful repressor of uricase, allantoinase, and allantoicase compared with ammonium. No pathway-specific induction is required for the synthesis of formamidase, uricase, allantoinase, and allantoicase. Altogether these features indicate that nitrogen metabolism inP. chrysosporium is similar to that inAspergillus nidulans in its regulation, despite the absence of pathway-specific induction of the enzymes examined. These results are consistent with the existence of a regulatory gene mediating nitrogen catabolite repression similar to theA. nidulans areA gene inP. chrysosporium. Although glycerol acts as a nonrepressive carbon source for lignin peroxidase production (except when used at high concentrations), glutamate totally represses lignin peroxidase even in cultures with glycerol. This indicates that carbon regulation and nitrogen regulation of lignin peroxidase may not be separated inP. chrysosporium.

Catabolism of arylglycerol-β-aryl ethers lignin model compounds by Pseudomonas cepacia 122

Pseudomonas cepacia 122 can grow on several lignin model compounds including the arylglycerol-beta-aryl ethers guaiacylglycerol-beta-coniferyl ether and guaiacylglycerol-beta-guaiacyl ether. Non-phenolic lignin model compounds are not degraded by this bacterium. The enzyme system catalyzing guaiacylglycerol-beta-guaiacyl ether dissimilation in Pseudomonas cepacia 122 is inducible and repressed by glucose. Guaiacylglycerol and guaiacylglycerol-beta-guaiacyl ether were identified as intermediates in guaiacylglycerol-beta-coniferyl ether catabolism. Guaiacol, guaiacoxyethanol, vanillin and vanillic acid were identified as intermediates of guaiacylglycerol-beta-guaiacyl ether breakdown indicating that a C alpha-C beta splitting mechanism is involved in the degradation of aryl-alkyl ethers by this bacterium.

Inheritance of cellulose- and lignin-degrading ability as well as endoglucanase isozyme pattern in Dichomitus squalens

SummaryThe progeny of Dichomitus squalens CBS-432-34 is heterogeneous with respect to specific growth rate on glucose, cellulolytic ([U14C]cellulose → 14CO2) and ligninolytic ([14C]synthetic lignin → 14CO2) activities with little correlation between these metric characters. Variations do not show clear-cut phenotypes but rather a continuous range between extreme values pointing to multigenic control of these characters. Most homocaryons showed decreased cellulolytic or ligninolytic activity compared to the parent dicaryon. However a few homocaryons were comparable or even superior to the parent dicaryon for ligninolytic or cellulolytic activity with no correlation between each factor. Strains with reduced cellulolytic activity and altered isozyme patterns of endoglucanases were isolated in the progeny of D. squalens CBS-432-34. While the parent strain produced three main endoglucanase multiple enzymes designated EnI, EnII and EnIII, several strains in the progeny produced a different multiple enzyme pattern. In contrast to the quantitative ability to degrade cellulose, multiple enzyme pattern variation in the progeny did not show continuous variations. characterization of heterocaryon phenotypes derived from Ien+ and Ien 1 homocaryons and first filial generation (f1) analysis showed that genetic control of the multiple enzyme pattern (Ien 1 phenotype) in D. squalens is complex.