Identification of a New Endo-β-1,4-xylanase Prospected from the Microbiota of the Termite <i>Heterotermes tenuis</i>
Xylanases are hemicellulases that break down xylan to soluble pentoses. They are used for industrial purposes, such as paper whitening, beverage clarification, and biofuel production. The second-generation bioethanol production is hindered by the enzymatic hydrolysis step of the lignocellulosic biom...
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MDPI AG
2022-04-01
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| author | Olinda S. A. Alcobaça Emeline B. Campanini Iara Ciancaglini Sâmara V. Rocha Iran Malavazi Caio C. M. Freire Francis M. F. Nunes Andrea S. C. Fuentes Anderson F. Cunha |
| author_facet | Olinda S. A. Alcobaça Emeline B. Campanini Iara Ciancaglini Sâmara V. Rocha Iran Malavazi Caio C. M. Freire Francis M. F. Nunes Andrea S. C. Fuentes Anderson F. Cunha |
| author_sort | Olinda S. A. Alcobaça |
| collection | DOAJ |
| description | Xylanases are hemicellulases that break down xylan to soluble pentoses. They are used for industrial purposes, such as paper whitening, beverage clarification, and biofuel production. The second-generation bioethanol production is hindered by the enzymatic hydrolysis step of the lignocellulosic biomass, due to the complex arrangement established among its constituents. Xylanases can potentially increase the production yield by improving the action of the cellulolytic enzyme complex. We prospected endo-β-1,4-xylanases from meta-transcriptomes of the termite <i>Heterotermes tenuis</i>. In silico structural characterization and functional analysis of an endo-β-1,4-xylanase from a symbiotic protist of <i>H. tenuis</i> indicate two active sites and a substrate-binding groove needed for the catalytic activity. No N-glycosylation sites were found. This endo-β-1,4-xylanase was recombinantly expressed in <i>Pichia pastoris</i> and <i>Escherichia coli</i> cells, presenting a molecular mass of approximately 20 kDa. Enzymatic activity assay using recombinant endo-β-1,4-xylanase was also performed on 1% xylan agar stained with Congo red at 30 °C and 40 °C. The enzyme expressed in both systems was able to hydrolyze the substrate xylan, becoming a promising candidate for further analysis aiming to determine its potential for application in industrial xylan degradation processes. |
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| spelling | doaj.art-3cbf528898fb4b44bc53a97be258549b2023-11-23T12:14:48ZengMDPI AGMicroorganisms2076-26072022-04-0110590610.3390/microorganisms10050906Identification of a New Endo-β-1,4-xylanase Prospected from the Microbiota of the Termite <i>Heterotermes tenuis</i>Olinda S. A. Alcobaça0Emeline B. Campanini1Iara Ciancaglini2Sâmara V. Rocha3Iran Malavazi4Caio C. M. Freire5Francis M. F. Nunes6Andrea S. C. Fuentes7Anderson F. Cunha8Department of Genetics and Evolution, Federal University of São Carlos, Sao Carlos 13565-905, BrazilDepartment of Genetics and Evolution, Federal University of São Carlos, Sao Carlos 13565-905, BrazilBrazilian Biorenewables National Laboratory, Brazilian Center of Research in Energy and Materials, Campinas 13083-100, BrazilDepartment of Genetics and Evolution, Federal University of São Carlos, Sao Carlos 13565-905, BrazilDepartment of Genetics and Evolution, Federal University of São Carlos, Sao Carlos 13565-905, BrazilDepartment of Genetics and Evolution, Federal University of São Carlos, Sao Carlos 13565-905, BrazilDepartment of Genetics and Evolution, Federal University of São Carlos, Sao Carlos 13565-905, BrazilDepartment of Genetics and Evolution, Federal University of São Carlos, Sao Carlos 13565-905, BrazilDepartment of Genetics and Evolution, Federal University of São Carlos, Sao Carlos 13565-905, BrazilXylanases are hemicellulases that break down xylan to soluble pentoses. They are used for industrial purposes, such as paper whitening, beverage clarification, and biofuel production. The second-generation bioethanol production is hindered by the enzymatic hydrolysis step of the lignocellulosic biomass, due to the complex arrangement established among its constituents. Xylanases can potentially increase the production yield by improving the action of the cellulolytic enzyme complex. We prospected endo-β-1,4-xylanases from meta-transcriptomes of the termite <i>Heterotermes tenuis</i>. In silico structural characterization and functional analysis of an endo-β-1,4-xylanase from a symbiotic protist of <i>H. tenuis</i> indicate two active sites and a substrate-binding groove needed for the catalytic activity. No N-glycosylation sites were found. This endo-β-1,4-xylanase was recombinantly expressed in <i>Pichia pastoris</i> and <i>Escherichia coli</i> cells, presenting a molecular mass of approximately 20 kDa. Enzymatic activity assay using recombinant endo-β-1,4-xylanase was also performed on 1% xylan agar stained with Congo red at 30 °C and 40 °C. The enzyme expressed in both systems was able to hydrolyze the substrate xylan, becoming a promising candidate for further analysis aiming to determine its potential for application in industrial xylan degradation processes.https://www.mdpi.com/2076-2607/10/5/906<i>Heterotermes tenuis</i>meta-transcriptomesymbiotic protisthemicellulasexylanasesecond-generation bioethanol |
| spellingShingle | Olinda S. A. Alcobaça Emeline B. Campanini Iara Ciancaglini Sâmara V. Rocha Iran Malavazi Caio C. M. Freire Francis M. F. Nunes Andrea S. C. Fuentes Anderson F. Cunha Identification of a New Endo-β-1,4-xylanase Prospected from the Microbiota of the Termite <i>Heterotermes tenuis</i> Microorganisms <i>Heterotermes tenuis</i> meta-transcriptome symbiotic protist hemicellulase xylanase second-generation bioethanol |
| title | Identification of a New Endo-β-1,4-xylanase Prospected from the Microbiota of the Termite <i>Heterotermes tenuis</i> |
| title_full | Identification of a New Endo-β-1,4-xylanase Prospected from the Microbiota of the Termite <i>Heterotermes tenuis</i> |
| title_fullStr | Identification of a New Endo-β-1,4-xylanase Prospected from the Microbiota of the Termite <i>Heterotermes tenuis</i> |
| title_full_unstemmed | Identification of a New Endo-β-1,4-xylanase Prospected from the Microbiota of the Termite <i>Heterotermes tenuis</i> |
| title_short | Identification of a New Endo-β-1,4-xylanase Prospected from the Microbiota of the Termite <i>Heterotermes tenuis</i> |
| title_sort | identification of a new endo β 1 4 xylanase prospected from the microbiota of the termite i heterotermes tenuis i |
| topic | <i>Heterotermes tenuis</i> meta-transcriptome symbiotic protist hemicellulase xylanase second-generation bioethanol |
| url | https://www.mdpi.com/2076-2607/10/5/906 |
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