Crystal structure determination of Pseudomonas stutzeri A1501 endoglucanase Cel5A: the search for a molecular basis for glycosynthesis in GH5_5 enzymes. Issue 6 (18th June 2019)

• Record Type: Journal Article
• Authors: Dutoit, Raphaël; Delsaute, Maud; Collet, Laetitia; Vander Wauven, Corinne; Van Elder, Dany; Berlemont, Renaud; Richel, Aurore; Galleni, Moreno; Bauvois, Cédric
• Is Part Of: Acta crystallographica. Volume 75:Issue 6(2019)
• Page Start: 605
• Page End: 615
• Publication Date: 2019-06-18
• Subjects: glycosyl hydrolase family 5 -- cellulases -- β‐1, 4‐endoglucanases -- TIM barrel -- Ps_Cel5A -- Pst_2494 -- Pseudomonas stutzeri -- transglycosylation; X-ray crystallography -- Periodicals; Crystallography -- Periodicals; Molecular biology -- Periodicals; Molecular structure -- Periodicals; Biomolecules -- Structure -- Periodicals; Cytology -- Periodicals; Biomolecules -- Structure; Crystallography; Cytology; Molecular biology; Molecular structure; X-ray crystallography; Periodicals; 548
• DOI: 10.1107/S2059798319007113
• Languages: English
• ISSNs: 2059-7983

Abstract

Abstract : The crystal structure of the endoglucanase Ps_Cel5A from Pseudomonas stutzeri A1501 revealed aglycone‐binding‐site adaptations that are possibly linked to its transglycosylation activity. Abstract : The discovery of new glycoside hydrolases that can be utilized in the chemoenzymatic synthesis of carbohydrates has emerged as a promising approach for various biotechnological processes. In this study, recombinant Ps_Cel5A from Pseudomonas stutzeri A1501, a novel member of the GH5_5 subfamily, was expressed, purified and crystallized. Preliminary experiments confirmed the ability of Ps_Cel5A to catalyze transglycosylation with cellotriose as a substrate. The crystal structure revealed several structural determinants in and around the positive subsites, providing a molecular basis for a better understanding of the mechanisms that promote and favour synthesis rather than hydrolysis. In the positive subsites, two nonconserved positively charged residues (Arg178 and Lys216) were found to interact with cellobiose. This adaptation has also been reported for transglycosylating β‐mannanases of the GH5_7 subfamily.