Insights into the dual cleavage activity of the GH16 laminarinase enzyme class on beta-1,3 and beta-1,4 glycosidic bonds

Autor(es):Mostrar menos -

Liberato, Marcelo Vizona ^{[1, 2]} ; Prates, Erica Teixeira ^{[3, 4, 5]} ; Goncalves, Thiago Augusto ^{[6, 2]} ; Bernardes, Amanda ^[7] ; Vilela, Nathalia ^{[6, 2]} ; Fattori, Juliana ^[8] ; Ematsu, Gabriela Cristina ^[1] ; Chinaglia, Mariana ^[1] ; Machi Gomes, Emerson Rodrigo ^[1] ; Migliorini Figueira, Ana Carolina ^[8] ; Damasio, Andre ^[6] ; Polikarpov, Igor ^[7] ; Skaf, Munir S. ^{[3, 5]} ; Squina, Fabio Marcio ^[2]

Afiliação do(s) autor(es)

^[1] Ctr Nacl Pesquisa Energia & Mat CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, Campinas, SP - Brazil

^[2] Univ Sorocaba, Programa Proc Tecnol & Ambientais, Sorocaba, SP - Brazil

^[3] Univ Estadual Campinas, Inst Quim, Campinas, SP - Brazil

^[4] Oak Ridge Natl Lab, Biosci Div, Oak Ridge, TN - USA

^[5] Univ Estadual Campinas, Ctr Pesquisa Engn & Ciencias Computacionais, Campinas, SP - Brazil

^[6] Univ Estadual Campinas, Inst Biol, Dept Bioquim & Biol Tecidual, Campinas, SP - Brazil

^[7] Univ Sao Paulo, Inst Fis Sao Carlos, Sao Carlos, SP - Brazil

^[8] Ctr Nacl Pesquisa Energia & Mat CNPEM, Lab Nacl Biociencias LNBio, Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Biological Chemistry; v. 296, JAN-JUN 2021.
• Citações Web of Science: 0

Resumo

Glycoside hydrolases (GHs) are involved in the degradation of a wide diversity of carbohydrates and present several biotechnological applications. Many GH families are composed of enzymes with a single well-defined specificity. In contrast, enzymes from the GH16 family can act on a range of different polysaccharides, including beta-glucans and galactans. SCLam, a GH16 member derived from a soil metagenome, an endo-beta 1,3(4)-glucanase (EC 3.2.1.6), can cleave both beta-1,3 and beta-1,4 glycosidic bonds in glucans, such as laminarin, barley beta-glucan, and cello-oligosaccharides. A similar cleavage pattern was previously reported for other GH16 family members. However, the molecular mechanisms for this dual cleavage activity on (1,3)- and (1,4)-beta-D-glycosidic bonds by laminarinases have not been elucidated. In this sense, we determined the X-ray structure of a presumably inactive form of SCLam cocrystallized with different oligosaccharides. The solved structures revealed general bound products that are formed owing to residual activities of hydrolysis and transglycosylation. Biochemical and biophysical analyses and molecular dynamics simulations help to rationalize differences in activity toward different substrates. Our results depicted a bulky aromatic residue near the catalytic site critical to select the preferable configuration of glycosidic bonds in the binding cleft. Altogether, these data contribute to understanding the structural basis of recognition and hydrolysis of beta-1,3 and beta-1,4 glycosidic linkages of the laminarinase enzyme class, which is valuable for future studies on the GH16 family members and applications related to biomass conversion into feedstocks and bioproducts. (AU)

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