Structures of BlEst2 from Bacillus licheniformis in its propeptide and mature forms reveal autoinhibitory effects of the C-terminal domain

Autor(es)

Nakamura, Aline Minali ; Godoy, Andre Schutzer ; Kadowaki, Marco Antonio Seiki ; Trentin, Lucas N. ; Gonzalez, Sinkler E. T. ; Skaf, Munir S. ; Polikarpov, Igor

• Tipo de documento: Artigo Científico
• Fonte: FEBS Journal; v. 291, n. 22, p. 21-pg., 2024-07-27.

Resumo

Carboxylesterases comprise a major class of alpha/beta-fold hydrolases responsible for the cleavage and formation of ester bonds. Found ubiquitously in nature, these enzymes are crucial for the metabolism of both endogenous and exogenous carboxyl esters in animals, plants and microorganisms. Beyond their essential physiological roles, carboxylesterases stand out as one of the important classes of biocatalysts for biotechnology. BlEst2, an enzyme previously classified as Bacillus licheniformis esterase, remains largely uncharacterized. In the present study, we elucidate the structural biology, molecular dynamics and biochemical features of BlEst2. Our findings reveal a canonical alpha/beta-hydrolase fold similar to the ESTHER block L of lipases, further augmented by two additional accessory C-terminal domains. Notably, the catalytic domain demonstrates two insertions, which occupy conserved locations in alpha/beta-hydrolase proteins and commonly form the lid domain in lipase structures. Intriguingly, our in vitro cleavage of C-terminal domains revealed the structure of the active form of BlEst2. Upon activation, BlEst2 showed a markedly elevated hydrolytic activity. This observation implies that the intramolecular C-terminal domain serves as a regulatory intramolecular inhibitor. Interestingly, despite exhibiting esterase-like activity, BlEst2 structural characteristics align more closely with lipases. This suggests that BlEst2 could potentially represent a previously unrecognized subgroup within the realm of carboxyl ester hydrolases. (AU)

• Processo FAPESP: 19/17373-0 - Estudos computacionais de enzimas ativas em carboidratos e interações de celulose com compostos de matriz de paredes celulares de plantas
• Beneficiário: Lucas Nascimento Trentin
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 13/08293-7 - CECC - Centro de Engenharia e Ciências Computacionais
• Beneficiário: Munir Salomao Skaf
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 21/08780-1 - Degradação enzimática de exopolissacharídeos de biofilmes microbianos: abordagens de biofísica estrutural, biotecnologia molecular e bioquímica sintética em estudos de CAZymes em busca de novas ferramentas enzimáticas para tratamentos antimicrobianos
• Beneficiário: Igor Polikarpov
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 19/17371-8 - Dinâmica molecular de enzimas para sacarificação glicanos
• Beneficiário: Sinkler Eduardo Tormet Gonzalez
• Modalidade de apoio: Bolsas no Brasil - Doutorado