Heterologous expression of lytic polysaccharide monooxygenases from Thermothelomyces thermophilus (2023)
- Authors:
- USP affiliated authors: SEGATO, FERNANDO - EEL ; MERCADO, MARTHA INES VELEZ - EEL ; BERTO, GABRIELA LEILA - EEL
- Unidade: EEL
- Subjects: MICROBIOLOGIA; BIOTECNOLOGIA
- Keywords: Thermothelomyces thermophilus; Heterologous expression; Lytic polysaccharide monooxygenase; Filamentous fungi; Aspergillus nidulans
- Agências de fomento:
- Language: Inglês
- Abstract: The thermophilic fungus Thermothelomyces thermophilus (previously Myceliophthora thermophila) has in its genome a significant number of genes encoding plant cell-wall-degrading enzymes. Since T. thermophilus can be a great source of thermostable proteins involved in the depolymerization of lignocellulosic materials different enzymes have been identified and produced from this fungus. The conversion of lignocellulose into its subunits is a limited step therefore, more studies are needed to improve this process. Recently, oxidative proteins that act directly on lignocellulosic biomass have been discovered and classified as lytic polysaccharide monooxygenases (LPMOs). Filamentous fungi contain large number of genes that codify to LPMOs as T. thermophilus which contains 23 codifying genes to LPMOs from the family AA9 (Auxiliary activity enzymes) and 3 codifying genes to LPMOs from the family AA16. Since T. thermophilus shows this considerable number of LPMOs in the genome and only a small number of them are commonly expressed, heterologous expression is important tool to over express and characterize these enzymes for their biotechnological application. This study aims to amplify and purify the LPMO’s codifying genes from T. thermophilus to be further introduced in Escherichia coli and over express in Aspergillus nidulans A773. Firstly, genes were amplified using specified oligonucleotides for each gene by PCR using T. thermophilus DNA as template. The PCR product was purified using the Wizard SV Gel and PCR Clean-Up System kit. Fragments of the genes were cloned into the plasmid pEXPYR by Gibson Assembly, and the plasmids were expanded in E. coli cells. Positive colonies were confirmed by PCR and cultivated on LB medium containing ampicillin. Colonies were incubated overnight and then, centrifugated to recover the pellet (cell biomass). DNA was extract from the positive colonies using the Wizard P
- Imprenta:
- Publisher: SBM
- Publisher place: Foz do Iguaçú-PR
- Date published: 2023
- Source:
- Título: Anais do 32o Congresso Brasileiro de Microbiologia
- ISSN: 2176-414X
- Volume/Número/Paginação/Ano: n.700-1, 2023
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ABNT
VELEZ-MERCADO, Martha I. e BERTO, Gabriela Leila e SEGATO, Fernando. Heterologous expression of lytic polysaccharide monooxygenases from Thermothelomyces thermophilus. 2023, Anais.. Foz do Iguaçú-PR: SBM, 2023. Disponível em: https://sbmicrobiologia.org.br/32cbm-anais/resumos/R-0700-1.html. Acesso em: 27 dez. 2025. -
APA
Velez-Mercado, M. I., Berto, G. L., & Segato, F. (2023). Heterologous expression of lytic polysaccharide monooxygenases from Thermothelomyces thermophilus. In Anais do 32o Congresso Brasileiro de Microbiologia. Foz do Iguaçú-PR: SBM. Recuperado de https://sbmicrobiologia.org.br/32cbm-anais/resumos/R-0700-1.html -
NLM
Velez-Mercado MI, Berto GL, Segato F. Heterologous expression of lytic polysaccharide monooxygenases from Thermothelomyces thermophilus [Internet]. Anais do 32o Congresso Brasileiro de Microbiologia. 2023 ;(700-1):[citado 2025 dez. 27 ] Available from: https://sbmicrobiologia.org.br/32cbm-anais/resumos/R-0700-1.html -
Vancouver
Velez-Mercado MI, Berto GL, Segato F. Heterologous expression of lytic polysaccharide monooxygenases from Thermothelomyces thermophilus [Internet]. Anais do 32o Congresso Brasileiro de Microbiologia. 2023 ;(700-1):[citado 2025 dez. 27 ] Available from: https://sbmicrobiologia.org.br/32cbm-anais/resumos/R-0700-1.html - Computational analysis of interaction between auxiliary activity enzymes from Thermothelomyces thermophilus
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