Production of Recombinant Bleaching Enzymes from Thermophilic Microorganisms in Fungal Hosts

Bergquist, Peter L.; Te’o, Junior; Gibbs, Moreland D.; Cziferszky, Angela; Faria, Fabrícia Paula de; Azevedo, M. O.; Nevalainen, Helena

doi:10.1385/abab:98-100:1-9:165

Cited by 15 publications

(8 citation statements)

References 29 publications

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“…Fungos do gênero Aspergillus têm sido escolhidos para processo de larga escala por serem capazes de produzirem grandes variedades e quantidades de enzimas em meios de baixo custo (Bergquist et al, 2002).…”

Section: Ligninaunclassified

Purificação e caracterização bioquímica e biofísica de uma xilanase de aspergillus foetidus

Cunha¹

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A minha orientadora, profa. Dra. Pérola de Oliveira Magalhães, por ter me recebido e me incentivado durante os dois anos de mestrado. Você foi uma referência profissional e pessoal para o meu crescimento. Obrigada por estar ao meu lado e acreditar em mim! Ao prof. Dr. Edivaldo Ximenes Ferreira Filho pela disponibilidade e atenção na resolução de dúvidas que surgiram durante a pesquisa. Pelo auxílio com os experimentos de cinética enzimática e de hidrólise. À profa. Dra. Sonia Maria de Freitas, pelos ensinamentos, paciência e oportunidade. Pelo auxílio nos experimentos de termodinâmica e de análise estrutural da xilanase. Ao prof. Dr. Thomas Christopher Rhys Williams, pelo auxílio no trabalho com o HPLC. A todos os professores do mestrado que de alguma forma contribuíram para a minha formação. Ao Programa de Pós-graduação em Ciências Farmacêuticas e toda sua equipe pela competência no suporte de nossas necessidades acadêmicas. A CAPES pelo apoio financeiro por meio de bolsa de estudos. A CAPES, CNPq, FAPDF, Finep, CT-Infra pelo apoio financeiro ao projeto de pesquisa. À Dra. Amanda Araújo Souza, pelos ensinamentos e vivências compartilhadas. Pelo auxílio nos experimentos de análise estrutural da xilanase e discussão dos resultados. À Dra. Leonora Rios de Souza Moreira e à Débora Lo Sciuto pelo auxílio nos experimentos de hidrólise e discussão dos resultados, pela paciência, disposição e ensinamentos. Ao Dr. Kleber Barros, pela disponibilidade e pelo auxílio na análise dos experimentos de planejamento fatorial. Ao Helder Andrey Rocha Gomes pelo ensinamento da técnica de ultrafiltração e pelas conversas que contribuíram para este trabalho. Aos colegas do Laboratório de Enzimologia-IB, que contribuíram de alguma forma para este trabalho. Às técnicas Michelle e Patrícia pelo apoio, carinho e ajuda, por tornarem os momentos no laboratório muito mais agradáveis. À amiga Raquel, pela amizade, companheirismo e pelos momentos que passamos juntas. Só nós sabemos o que passamos e o quanto conversamos nesse momento juntas. Você foi meu apoio e inspiração durante o mestrado.

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Section: Ligninaunclassified

Purificação e caracterização bioquímica e biofísica de uma xilanase de aspergillus foetidus

Cunha¹

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“…They also have a strong survivability, which make them an ideal cell factory for producing drugs, antibiotics, industrial enzymes, and other substances. Most importantly, filamentous fungi have a strong ability in protein expression and perform various posttranslational processing correctly, including glycosylation, peptide chain shearing, and disulfide bond formation, which are similar to mammal cells ( Bergquist et al, 2002 ). Besides, filamentous fungi have a powerful secretory pathway, including signal recognition particle signaling and efficient function of the endoplasmic reticulum in protein modification, as well as rapid clearance of misfolded proteins, fusion between vesicles and target membranes, and apical secretion of the proteins, which conferred them the ability to produce eukaryotic proteins correctly ( Kavanagh, 2011 ; Karagiosis and Baker, 2012 ; Fang and Qu, 2018 ).…”

Section: Introductionmentioning

confidence: 99%

Regulating Strategies for Producing Carbohydrate Active Enzymes by Filamentous Fungal Cell Factories

Zhang

Liu

et al. 2020

Front. Bioeng. Biotechnol.

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Filamentous fungi are important eukaryotic organisms crucial in substrate degradation and carbon cycle on the earth and have been harnessed as cell factories for the production of proteins and other high value-added products in recent decades. As cell factories, filamentous fungi play a crucial role in industrial protein production as both native hosts and heterologous hosts. In this review, the regulation strategies of carbohydrate active enzyme expression at both transcription level and protein level are introduced, and the transcription regulations are highlighted with induction mechanism, signaling pathway, and promoter and transcription factor regulation. Afterward, the regulation strategies in protein level including suitable posttranslational modification, protein secretion enhancement, and protease reduction are also presented. Finally, the challenges and perspectives in this field are discussed. In this way, a comprehensive knowledge regarding carbohydrate active enzyme production regulation at both transcriptional and protein levels is provided with the particular goal of aiding in the practical application of filamentous fungi for industrial protein production.

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“…These residues could be utilized by filamentous fungi as a carbon source for the production of enzymes, in particular hydrolytic ones (Moreira et al 2012). Aspergillus fungi have been chosen for large-scale processes because they can produce large quantities and varieties of enzymes in a low-cost medium (Bergquist et al 2002). Moreira et al (2012) studied the degradation of lignocellulosic residues for production of enzymes of industrial significance such as xylanases, mannanases, pectinases, β-glucosidases, avicelases, phosphatases, and carboxymethyl cellulases by different species of fungi isolated from soil, including Aspergillusterreus , Aspergillusoryzae , and Aspergillusniger (Moreira et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Optimization and partial purification of beta-galactosidase production by Aspergillus niger isolated from Brazilian soils using soybean residue

et al. 2019

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β-Galactosidases are widely used for industrial applications. These enzymes could be used in reactions of lactose hydrolysis and transgalactosylation. The objective of this study was the production, purification, and characterization of an extracellular β-galactosidase from a filamentous fungus, Aspergillus niger. The enzyme production was optimized by a factorial design. Maximal β-galactosidase activity (24.64 U/mL) was found in the system containing 2% of a soybean residue (w/v) at initial pH 7.0, 28 °C, 120 rpm in 7 days. ANOVA of the optimization study indicated that the response data on temperature and pH were significant (p < 0.05). The regression equation indicated that the R 2 is 0.973. Ultrafiltration at a 100 and 30 kDa cutoff followed by gel filtration and anion exchange chromatography were carried out to purify the fungal β-galactosidase. SDS-PAGE revealed a protein with molecular weight of approximately 76 kDa. The partially purified enzyme showed an optimum temperature of 50 °C and optimum pH of 5.0, being stable under these conditions for 15 h. The enzyme was exposed to conditions approaching gastric pH and in pepsin’s presence, 80% of activity was preserved after 2 h. These results reveal a A. niger β-galactosidase obtained from residue with favorable characteristics for food industries.

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Production of Recombinant Bleaching Enzymes from Thermophilic Microorganisms in Fungal Hosts

Cited by 15 publications

References 29 publications

Purificação e caracterização bioquímica e biofísica de uma xilanase de aspergillus foetidus

Purificação e caracterização bioquímica e biofísica de uma xilanase de aspergillus foetidus

Regulating Strategies for Producing Carbohydrate Active Enzymes by Filamentous Fungal Cell Factories

Optimization and partial purification of beta-galactosidase production by Aspergillus niger isolated from Brazilian soils using soybean residue

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