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Insights into the xylan degradation system of Cellulomonas sp. B6: biochemical characterization of rCsXyn10A and rCsAbf62A.
Garrido, Mercedes María; Piccinni, Florencia Elizabeth; Landoni, Malena; Peña, María Jesús; Topalian, Juliana; Couto, Alicia; Wirth, Sonia Alejandra; Urbanowicz, Breeanna Rae; Campos, Eleonora.
Afiliação
  • Garrido MM; Instituto de Agrobiotecnología y Biología Molecular (IABIMO), CICVyA, Instituto Nacional de Tecnología Agropecuaria (INTA)- CONICET, Los Reseros y Nicolás Repetto S/N (1686), Hurlingham, Buenos Aires, Argentina.
  • Piccinni FE; Laboratorio de Agrobiotecnología, DFBMC- FCEN and Instituto de Biodiversidad y Biología Experimental y Aplicada (IBBEA) CONICET- Universidad de Buenos Aires (UBA), Pab. II, Ciudad Universitaria, C1428EG, Buenos Aires, Argentina.
  • Landoni M; Instituto de Agrobiotecnología y Biología Molecular (IABIMO), CICVyA, Instituto Nacional de Tecnología Agropecuaria (INTA)- CONICET, Los Reseros y Nicolás Repetto S/N (1686), Hurlingham, Buenos Aires, Argentina.
  • Peña MJ; Laboratorio de Agrobiotecnología, DFBMC- FCEN and Instituto de Biodiversidad y Biología Experimental y Aplicada (IBBEA) CONICET- Universidad de Buenos Aires (UBA), Pab. II, Ciudad Universitaria, C1428EG, Buenos Aires, Argentina.
  • Topalian J; Centro de Investigación en Hidratos de Carbono (CIHIDECAR)- CONICET, Departamento de Química Orgánica, FCEN- Universidad de Buenos Aires (UBA), Pab. II, Ciudad Universitaria, C1428EG, Buenos Aires, Argentina.
  • Couto A; Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Road, Athens, GA, USA.
  • Wirth SA; Instituto de Agrobiotecnología y Biología Molecular (IABIMO), CICVyA, Instituto Nacional de Tecnología Agropecuaria (INTA)- CONICET, Los Reseros y Nicolás Repetto S/N (1686), Hurlingham, Buenos Aires, Argentina.
  • Urbanowicz BR; Centro de Investigación en Hidratos de Carbono (CIHIDECAR)- CONICET, Departamento de Química Orgánica, FCEN- Universidad de Buenos Aires (UBA), Pab. II, Ciudad Universitaria, C1428EG, Buenos Aires, Argentina.
  • Campos E; Laboratorio de Agrobiotecnología, DFBMC- FCEN and Instituto de Biodiversidad y Biología Experimental y Aplicada (IBBEA) CONICET- Universidad de Buenos Aires (UBA), Pab. II, Ciudad Universitaria, C1428EG, Buenos Aires, Argentina.
Appl Microbiol Biotechnol ; 106(13-16): 5035-5049, 2022 Aug.
Article em En | MEDLINE | ID: mdl-35799069
Valorization of the hemicellulose fraction of plant biomass is crucial for the sustainability of lignocellulosic biorefineries. The Cellulomonas genus comprises Gram-positive Actinobacteria that degrade cellulose and other polysaccharides by secreting a complex array of enzymes. In this work, we studied the specificity and synergy of two enzymes, CsXyn10A and CsAbf62A, which were identified as highly abundant in the extracellular proteome of Cellulomonas sp. B6 when grown on wheat bran. To explore their potential for bioprocessing, the recombinant enzymes were expressed and their activities were thoroughly characterized. rCsXyn10A is a GH10 endo-xylanase (EC 3.2.1.8), active across a broad pH range (5 to 9), at temperatures up to 55 °C. rCsAbf62A is an α-L-arabinofuranosidase (ABF) (EC 3.2.1.55) that specifically removes α-1,2 and α-1,3-L-arabinosyl substituents from arabino-xylo-oligosaccharides (AXOS), xylan, and arabinan backbones, but it cannot act on double-substituted residues. It also has activity on pNPA. No differences were observed regarding activity when CsAbf62A was expressed with its appended CBM13 module or only the catalytic domain. The amount of xylobiose released from either wheat arabinoxylan or arabino-xylo-oligosaccharides increased significantly when rCsXyn10A was supplemented with rCsAbf62A, indicating that the removal of arabinosyl residues by rCsAbf62A improved rCsXyn10A accessibility to ß-1,4-xylose linkages, but no synergism was observed in the deconstruction of wheat bran. These results contribute to designing tailor-made, substrate-specific, enzymatic cocktails for xylan valorization. KEY POINTS: • rCsAbf62A removes α-1,2 and α-1,3-L-arabinosyl substituents from arabino-xylo-oligosaccharides, xylan, and arabinan backbones. • The appended CBM13 of rCsAbf62A did not affect the specific activity of the enzyme. • Supplementation of rCsXyn10A with rCsAbf62A improves the degradation of AXOS and xylan.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Xilanos / Cellulomonas Idioma: En Revista: Appl Microbiol Biotechnol Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Argentina País de publicação: Alemanha

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Xilanos / Cellulomonas Idioma: En Revista: Appl Microbiol Biotechnol Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Argentina País de publicação: Alemanha