Engineering Bifunctional Laccase-Xylanase Chimeras for Improved Catalytic Performance

Ribeiro, Lucas F.; Furtado, Gilvan Pessoa; Lourenzoni, Marcos Roberto; Costa-Filho, Antonio J.; Santos, C.R.; Nogueira, Simone C. Peixoto; Betini, Jorge A.; Polizeli, Maria de Lourdes Teixeira de Moraes; Murakami, Mário Tyago; Ward, Richard J.

doi:10.1074/jbc.m111.253419

Cited by 60 publications

(39 citation statements)

References 68 publications

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“…Moilanen et al (2011) employed blend of commercial cellulases and laccases on pretreated spruce and obtained 12% increase in hydrolysis yield. Furtado et al (2013) and Ribeiro et al (2011) have also demonstrated the improvement in synergy and catalytic performance of fused laccases-(hemi)cellulase complex for biomass hydrolysis.…”

Section: Biological Additivesmentioning

confidence: 99%

“…A critical factor for success in the creation of enzyme chimeras is the compatibility and cooperativity among the involved CBMs and CDs, with respect to their physicochemical requirements such as solubility, optimum pH, and temperature Ribeiro et al 2011). Howbeit, Stern et al (2015) suggested that the optimal order for the positioning of enzymes as per their investigation is processive endoglucanase, exoglucanase, and non-processive endoglucanase; and for overall higher enzymatic activity the CBM should not be placed in the middle of the scaffoldin.…”

Section: Cell-surface Display Systemsmentioning

confidence: 99%

“…They reported that the different architectures of the bifunctional enzymes displayed "somewhat inferior" cellulolytic activity to that of the wild-type free enzyme system. However, Ribeiro et al (2011) created two bifunctional enzymes with xylanase-laccase activity using rational design methods and reported catalytic properties similar to the parental enzymes. Moreover, Chang et al (2011) reported an excellent performance of a bifunctional xylanase/endoglucanase (RuCelA), which distinguishes it as an ideal candidate for industrial applications.…”

Section: Multifunctionalmentioning

confidence: 99%

“…Engineering multi-domain enzymes that are capable of catalyzing two or more reactions is a potential strategy to reduce enzyme costs in bio-industrial processes, as multiple catalytic properties on a single polypeptide conceivably simplify production and purification operations of biochemicals (Ribeiro et al 2011). Similarly, the cost-effective optimization of chimeras to prevent unproductive competition between functionally similar enzymes by testing the importance of both the positions of enzymes and CBMs for an efficient use in bioprocessing industries is necessary, though demanding a vigorous investigation.…”

Section: Cell-surface Display Systemsmentioning

confidence: 99%

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Lignocellulases: a review of emerging and developing enzymes, systems, and practices

Obeng

Adam

Budiman

et al. 2017

Bioresour. Bioprocess.

132

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The highly acclaimed prospect of renewable lignocellulosic biocommodities as obvious replacement of their fossilbased counterparts is burgeoning within the last few years. However, the use of the abundant lignocellulosic biomass provided by nature to produce value-added products, especially bioethanol, still faces significant challenges. One of the crucial challenging factors is in association with the expression levels, stability, and cost-effectiveness of the cellulose-degrading enzymes (cellulases). Interestingly, several recommendable endeavors in the bid to curb these challenges are in pursuance. However, the existing body of literature has not well provided the updated roadmap of the advancement and key players spearheading the current success. Moreover, the description of enzyme systems and emerging paradigms with high prospects, for example, the cell-surface display system has been ill-captured in the literature. This review focuses on the lignocellulosic biocommodity pathway, with emphasis on cellulase and hemicellulase systems. The paradigm shift towards cell-surface display system and its emerging recommendable developments have also been discussed. The attempts in supplementing cellulase with other enzymes, accessory proteins, and chemical additives have also been discussed. Moreover, some of the prominent and influential discoveries in the cellulase fraternity have been discussed.

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Section: Biological Additivesmentioning

confidence: 99%

Section: Cell-surface Display Systemsmentioning

confidence: 99%

Section: Multifunctionalmentioning

confidence: 99%

Section: Cell-surface Display Systemsmentioning

confidence: 99%

See 2 more Smart Citations

Lignocellulases: a review of emerging and developing enzymes, systems, and practices

Obeng

Adam

Budiman

et al. 2017

Bioresour. Bioprocess.

132

View full text Add to dashboard Cite

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“…Além disso, quando as enzimas são fundidas, a proximidade física entre os domínios catalíticos pode aumentar a velocidade das reações, e frequentemente essas enzimas têm um melhor desempenho em comparação com as atividades das enzimas parentais (FONSECA-MALDONADO et al, 2014;LOURENZONI, 2013;FURTADO et al, 2013b;RIBEIRO et al, 2011a). Um aspecto fundamental para a construção de quimeras é a preservação ou melhoria das características catalíticas em relação às proteínas parentais (ADLAKHA et al, 2011).…”

Section: Produção De Enzimas Multidomíniosunclassified

Criação de uma enzima multifuncional feruloil esterase/acetil-xilano esterase por desenho racional

Alves¹

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AGRADECIMENTOSMuitas foram as pessoas que contribuíram, de modo especial (e único), para a conclusão deste trabalho. Com ensinamentos, sugestões, críticas, puxões de orelha, ajudas na bancada... Com a convivência agradável e divertida ao longo dessa caminhada... Com a amizade, carinho e presença confortante nos momentos difíceis... A Deus, causa primária de todas as coisas, que me dá força e plenas condições.Aos meus pais, agradeço pelo amor incondicional, pela paciência, pelo carinho, pelo incentivo... Obrigada por sempre sonharem junto comigo, sem medir esforços! Amo vocês! Ló, obrigada por ser uma irmã tão amiga! Pelas conversas, pela cumplicidade, pelo carinho e pelos momentos agradáveis em casa. from Clostridium thermocellum were fused to generate a bifunctional chimera feruloyl esterase/acetylxylan esterase (FaeAxe). A molecular model was created by rational design using a 3D-structure guided strategy. The fusion was created using overlap PCR, and the resulting product was cloned into the pETSUMO vector. The chimeric protein and the parental enzymes were expressed in Escherichia coli and purified and the enzymes were expressed in soluble form. Xylanases, feruloyl esterases and acetylxylan esterases degrade arabinoxylan polymers and their activity is a key step in the saccharification of biomass. The catalytic properties of the chimera and of the parental enzymes were tested against wheat and sugarcane arabinoxylan polymers after hydrolysis by GH11 endoxylanase from Bacillus subtilis and analyzed by mass spectroscopy. The deacetylase activity of acetyl-xylan esterase parental enzyme and FaeAxe chimera were confirmed, showing that the chimera kept the deacetylase activity. After hydrolysis by GH11 endoxylanase from Bacillus subtilis the feruloyl esterase and FaeAxe chimera activities on ferulic acid removal were not observed.

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Mapping secondary substrate‐binding sites on the GH11 xylanase from Bacillus subtilis

Molina,

Mendes,

Fuzo

et al. 2024

FEBS Letters

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Xylanases are of significant interest for biomass conversion technologies. Here, we investigated the allosteric regulation of xylan hydrolysis by the Bacillus subtilis GH11 endoxylanase. Molecular dynamics simulations (MDS) in the presence of xylobiose identified binding to the active site and two potential secondary binding sites (SBS) around surface residues Asn54 and Asn151. Arabinoxylan titration experiments with single cysteine mutants N54C and N151C labeled with the thiol‐reactive fluorophore acrylodan or the ESR spin‐label MTSSL validated the MDS results. Ligand binding at the SBS around Asn54 confirms previous reports, and analysis of the second SBS around N151C discovered in the present study includes residues Val98/Ala192/Ser155/His156. Understanding the regulation of xylanases contributes to efforts for industrial decarbonization and to establishing a sustainable energy matrix.

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Engineering Bifunctional Laccase-Xylanase Chimeras for Improved Catalytic Performance

Cited by 60 publications

References 68 publications

Lignocellulases: a review of emerging and developing enzymes, systems, and practices

Lignocellulases: a review of emerging and developing enzymes, systems, and practices

Criação de uma enzima multifuncional feruloil esterase/acetil-xilano esterase por desenho racional

Mapping secondary substrate‐binding sites on the GH11 xylanase from Bacillus subtilis

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