Background: 1,3-beta-glucan is a polysaccharide widely distributed in the cell wall of several phylogenetically distant organisms, such as bacteria, fungi, plants and microalgae. The presence of highly active 1,3-beta-glucanases in fungi evokes the biological question on how these organisms can efficiently metabolize exogenous sources of 1,3-beta-glucan without incurring in autolysis.Results: To elucidate the molecular mechanisms at the basis of 1,3-beta-glucan metabolism in fungal saprotrophs, the putative exo-1,3-beta-glucanase G9376 and a truncated form of the putative glucan endo-1,3-beta-glucosidase (Delta G7048) from Penicillium sumatraense AQ67100 were heterologously expressed in Pichia pastoris and characterized both in terms of activity and structure. G9376 efficiently converted laminarin and 1,3-beta-glucan oligomers into glucose by acting as an exo-glycosidase, whereas G7048 displayed a 1,3-beta-transglucanase/branching activity toward 1,3-beta-glucan oligomers with a degree of polymerization higher than 5, making these oligomers more recalcitrant to the hydrolysis acted by exo-1,3-beta-glucanase G9376. The X-ray crystallographic structure of the catalytic domain of G7048, solved at 1.9 angstrom of resolution, consists of a (beta/alpha)8 TIM-barrel fold characteristic of all the GH17 family members. The catalytic site is in a V-shaped cleft containing the two conserved catalytic glutamic residues. Molecular features compatible with the activity of G7048 as 1,3-beta-transglucanase are discussed.Conclusions: The antagonizing activity between AG7048 and G9376 indicates how opportunistic fungi belonging to Penicillium genus can feed on substrates similar for composition and structure to their own cell wall without incurring in a self-deleterious autohydrolysis.

Characterization of two 1,3-beta-glucan-modifying enzymes from Penicillium sumatraense reveals new insights into 1,3-beta-glucan metabolism of fungal saprotrophs

Scafati, V;Ponziani, S;Scortica, A;Di Matteo, A;
2022-01-01

Abstract

Background: 1,3-beta-glucan is a polysaccharide widely distributed in the cell wall of several phylogenetically distant organisms, such as bacteria, fungi, plants and microalgae. The presence of highly active 1,3-beta-glucanases in fungi evokes the biological question on how these organisms can efficiently metabolize exogenous sources of 1,3-beta-glucan without incurring in autolysis.Results: To elucidate the molecular mechanisms at the basis of 1,3-beta-glucan metabolism in fungal saprotrophs, the putative exo-1,3-beta-glucanase G9376 and a truncated form of the putative glucan endo-1,3-beta-glucosidase (Delta G7048) from Penicillium sumatraense AQ67100 were heterologously expressed in Pichia pastoris and characterized both in terms of activity and structure. G9376 efficiently converted laminarin and 1,3-beta-glucan oligomers into glucose by acting as an exo-glycosidase, whereas G7048 displayed a 1,3-beta-transglucanase/branching activity toward 1,3-beta-glucan oligomers with a degree of polymerization higher than 5, making these oligomers more recalcitrant to the hydrolysis acted by exo-1,3-beta-glucanase G9376. The X-ray crystallographic structure of the catalytic domain of G7048, solved at 1.9 angstrom of resolution, consists of a (beta/alpha)8 TIM-barrel fold characteristic of all the GH17 family members. The catalytic site is in a V-shaped cleft containing the two conserved catalytic glutamic residues. Molecular features compatible with the activity of G7048 as 1,3-beta-transglucanase are discussed.Conclusions: The antagonizing activity between AG7048 and G9376 indicates how opportunistic fungi belonging to Penicillium genus can feed on substrates similar for composition and structure to their own cell wall without incurring in a self-deleterious autohydrolysis.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11575/127978
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