Substrate and product hydrolysis specificity in family 11 glycoside hydrolases: an analysis of Penicillium funiculosum and Penicillium griseofulvum xylanases

Berrin, Jean‐Guy; Ajandouz, El Hassan; Georis, Jacques; Arnaut, Filip; Juge, Nathalie

doi:10.1007/s00253-006-0764-0

Cited by 54 publications

(45 citation statements)

References 17 publications

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“…This family module, formerly designated cellulosebinding domain family 1 (CBD1), is a well-characterized module specific to cellulose and exclusively in fungi (12). PaXyn11A xylanase exhibited an "endo" mode of action on polymeric substrates, releasing mainly xylobiose as major end product with a high catalytic efficiency, a common characteristic of fungal GH11 xylanases (4,11).…”

Section: Discussionmentioning

confidence: 99%

Podospora anserina Hemicellulases Potentiate the Trichoderma reesei Secretome for Saccharification of Lignocellulosic Biomass

Couturier

Haon²,

Coutinho

et al. 2011

Appl Environ Microbiol

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To improve the enzymatic hydrolysis (saccharification) of lignocellulosic biomass by Trichoderma reesei, a set of genes encoding putative polysaccharide-degrading enzymes were selected from the coprophilic fungus Podospora anserina using comparative genomics. Five hemicellulase-encoding genes were successfully cloned and expressed as secreted functional proteins in the yeast Pichia pastoris. These novel fungal CAZymes belonging to different glycoside hydrolase families (PaMan5A and PaMan26A mannanases, PaXyn11A xylanase, and PaAbf51A and PaAbf62A arabinofuranosidases) were able to break down their predicted cognate substrates. Although PaMan5A and PaMan26A displayed similar specificities toward a range of mannan substrates, they differed in their end products, suggesting differences in substrate binding. The N-terminal CBM35 module of PaMan26A displayed dual binding specificity toward xylan and mannan. PaXyn11A harboring a C-terminal CBM1 module efficiently degraded wheat arabinoxylan, releasing mainly xylobiose as end product. PaAbf51A and PaAbf62A arabinose-debranching enzymes exhibited differences in activity toward arabinose-containing substrates. Further investigation of the contribution made by each P. anserina auxiliary enzyme to the saccharification of wheat straw and spruce demonstrated that the endo-acting hemicellulases (PaXyn11A, PaMan5A, and PaMan26A) individually supplemented the secretome of the industrial T. reesei CL847 strain. The most striking effect was obtained with PaMan5A that improved the release of total sugars by 28% and of glucose by 18%, using spruce as lignocellulosic substrate.Lignocellulosic biomass is a desirable feedstock for the supply of second-generation bioethanol, being the largest renewable source of carbohydrates, but the recalcitrance of raw materials makes biotechnological conversion complex and costly (34).The main components of plant cell walls are cellulose, lignin, and hemicellulose, which form a tight complex with varying proportions between plants (for a review, see reference 45). Cellulose microfibrils are formed by crystalline-like combination of linear and highly ordered polymer chains of ␤-1,4-linked D-glucose residues, and lignin consists of an amorphous aromatic heteropolymer with hydrophobic properties. Hemicellulose is a combined designation of a diverse set of noncrystalline carbohydrate polymers from plant tissues that form closely associated networks with cellulose microfibrils and lignin (6, 24). Typically, the most abundant component of hemicellulose in the cell walls of monocots (e.g., cereals) is ␤-1,4-xylan, which consists of ␤-1,4-linked D-xylose residues substituted with L-arabinosyl, 4-O-methyl-glucuronosyl and acetyl side chains (46,53). Arabinofuranosyl residues of arabinoxylan may also be esterified with hydroxycinnamic acid residues, e.g., ferulic and p-coumaric acids (53). In softwoods (gymnosperms), the main hemicellulose component is ␤-1,4-mannan, which consists of a backbone formed by ␤-1,4-linked D-mannose and D-glucose residues substitu...

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Section: Discussionmentioning

confidence: 99%

Podospora anserina Hemicellulases Potentiate the Trichoderma reesei Secretome for Saccharification of Lignocellulosic Biomass

Couturier

Haon²,

Coutinho

et al. 2011

Appl Environ Microbiol

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“…Results are expressed in mol of sugar released per min per mg of enzyme. The specificity constants were calculated using the Matsui equation for oligosaccharides (2,16). For determination of Michaelis-Menten constants on laminarin and pustulan, the initial velocities were measured for substrate concentrations ranging from 1.25 to 20 mg ml Ϫ1 .…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Broad-Specificity β-Glucanase Acting on β-(1,3)-, β-(1,4)-, and β-(1,6)-Glucans That Defines a New Glycoside Hydrolase Family

Lafond

Navarro

Haon

et al. 2012

Appl Environ Microbiol

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Here we report the cloning of the Pa_3_10940 gene from the coprophilic fungus Podospora anserina, which encodes a C-terminal family 1 carbohydrate binding module (CBM1) linked to a domain of unknown function. The function of the gene was investigated by expression of the full-length protein and a truncated derivative without the CBM1 domain in the yeast Pichia pastoris. Using a library of polysaccharides of different origins, we demonstrated that the full-length enzyme displays activity toward a broad range of ␤-glucan polysaccharides, including laminarin, curdlan, pachyman, lichenan, pustulan, and cellulosic derivatives. Analysis of the products released from polysaccharides revealed that this ␤-glucanase is an exo-acting enzyme on ␤-(1,3)-and ␤-(1,6)-linked glucan substrates and an endo-acting enzyme on ␤-(1,4)-linked glucan substrates. Hydrolysis of short ␤-(1,3), ␤-(1,4), and ␤-(1,3)/␤-(1,4) gluco-oligosaccharides confirmed this striking feature and revealed that the enzyme performs in an exo-type mode on the nonreducing end of gluco-oligosaccharides. Excision of the CBM1 domain resulted in an inactive enzyme on all substrates tested. To our knowledge, this is the first report of an enzyme that displays bifunctional exo-␤-(1,3)/(1,6) and endo-␤-(1,4) activities toward beta-glucans and therefore cannot readily be assigned to existing Enzyme Commission groups. The amino acid sequence has high sequence identity to hypothetical proteins within the fungal taxa and thus defines a new family of glycoside hydrolases, the GH131 family.

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“…All the assays were carried out in duplicate. The data were fitted to the equation of Matsui (29,30) O and 4% from the enzyme and substrate stock solutions) containing 1 mM sodium acetate buffer, pH 5, 0.8 mM substrate, and 0.1 M enzyme. Samples (0.5 l) were withdrawn at different time points (0 -60 min) and spotted directly on a stainless steel plate for matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) analysis.…”

Section: Hydrolysis Product Formation From Oligosaccharides and Determentioning

confidence: 99%

Structural and Biochemical Analyses of Glycoside Hydrolase Families 5 and 26 β-(1,4)-Mannanases from Podospora anserina Reveal Differences upon Manno-oligosaccharide Catalysis

Couturier

Roussel

Rosengren

et al. 2013

Journal of Biological Chemistry

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121

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Background: Fungal mannanases contribute to enzymatic degradation of lignocellulose. Results: New fungal mannanases reveal striking differences in substrate specificities. A rigid linker tightly connects the family 26 glycoside hydrolase to its binding module. Conclusion: Podospora anserina mannanases display differences in substrate binding modes, transglycosylation activity, and modular organization. Significance: Information on the structure-function relationships of fungal mannanases is essential to improve the comprehension of biomass deconstruction.

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Substrate and product hydrolysis specificity in family 11 glycoside hydrolases: an analysis of Penicillium funiculosum and Penicillium griseofulvum xylanases

Cited by 54 publications

References 17 publications

Podospora anserina Hemicellulases Potentiate the Trichoderma reesei Secretome for Saccharification of Lignocellulosic Biomass

Podospora anserina Hemicellulases Potentiate the Trichoderma reesei Secretome for Saccharification of Lignocellulosic Biomass

Characterization of a Broad-Specificity β-Glucanase Acting on β-(1,3)-, β-(1,4)-, and β-(1,6)-Glucans That Defines a New Glycoside Hydrolase Family

Structural and Biochemical Analyses of Glycoside Hydrolase Families 5 and 26 β-(1,4)-Mannanases from Podospora anserina Reveal Differences upon Manno-oligosaccharide Catalysis

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