Glucomannan and beta-glucan degradation by Mytilus edulis Cel45A: Crystal structure and activity comparison with GH45 subfamily A, B and C. (1st February 2022)

• Record Type: Journal Article
• Authors: Okmane, Laura; Nestor, Gustav; Jakobsson, Emma; Xu, Bingze; Igarashi, Kiyohiko; Sandgren, Mats; Kleywegt, Gerard J.; Ståhlberg, Jerry
• Is Part Of: Carbohydrate polymers. Volume 277(2022)
• Publication Date: 2022-02-01
• Subjects: Endoglucanase -- Blue mussel -- Cel45A -- GH45 -- Beta-glucan -- Glucomannan; AcCel45A Ampullaria crossean Cel45A -- BG betaglucan -- CBM carbohydrate binding module -- CMC carboxymethyl cellulose -- DPBB double psi beta barrel -- GH glycoside hydrolase -- GH45 glycoside hydrolase family 45 -- GM glucomannan -- HiCel45A Humicola insolens Cel45A -- MeCel45A Mytilus edulis Cel45A -- PASC phosphoric acid swollen cellulose -- PcCel45A Phanerochaete chrysosporium Cel45A -- PHBAH p-Hydroxybenzoic acid hydrazide -- RMSD root mean square deviation -- TrCel45A Trichoderma reesei Cel45A; Polysaccharides -- Periodicals; Polysaccharides -- Periodicals; Polysaccharides -- Périodiques; Electronic journals; 547.78
• DOI: 10.1016/j.carbpol.2021.118771
• Languages: English
• ISSNs: 0144-8617

Abstract

Abstract: The enzymatic hydrolysis of barley beta-glucan, konjac glucomannan and carboxymethyl cellulose by a β-1, 4-D-endoglucanase MeCel45A from blue mussel, Mytilus edulis, which belongs to subfamily B of glycoside hydrolase family 45 (GH45), was compared with GH45 members of subfamilies A ( Humicola insolens HiCel45A), B ( Trichoderma reesei TrCel45A) and C ( Phanerochaete chrysosporium PcCel45A). Furthermore, the crystal structure of MeCel45A is reported. Initial rates and hydrolysis yields were determined by reducing sugar assays and product formation was characterized using NMR spectroscopy. The subfamily B and C enzymes exhibited mannanase activity, whereas the subfamily A member was uniquely able to produce monomeric glucose. All enzymes were confirmed to be inverting glycoside hydrolases. MeCel45A appears to be cold adapted by evolution, as it maintained 70% activity on cellohexaose at 4 °C relative to 30 °C, compared to 35% for TrCel45A. Both enzymes produced cellobiose and cellotetraose from cellohexaose, but TrCel45A additionally produced cellotriose. Graphical abstract: Unlabelled Image Highlights: GH45 subfamily A, B and C members are inverting enzymes. GH45 subfamily B and C members exhibit mannanase activity on glucomannan. Cel45A possibly adapted to cold in northern mollusc. Differences in linkage preference on glucomannan among GH45 subfamilies A, B and C