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Expression, Docking, and Molecular Dynamics of Endo-β-1,4-xylanase I Gene of Trichoderma virens in Pichia stipitis
It is essential that major carbohydrate polymers in the lignocellulosic biomass are converted into fermentable sugars for the economical production of energy. Xylan, the major component of hemicelluloses, is the second most naturally abundant carbohydrate polymer comprising 20–40% of the total bioma...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Hindawi
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5569632/ https://www.ncbi.nlm.nih.gov/pubmed/28856159 http://dx.doi.org/10.1155/2017/4658584 |
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author | Wickramasinghe, Gammadde Hewa Ishan Maduka Rathnayake, Pilimathalawe Panditharathna Attanayake Mudiyanselage Samith Indika Chandrasekharan, Naduviladath Vishvanath Weerasinghe, Mahindagoda Siril Samantha Wijesundera, Ravindra Lakshman Chundananda Wijesundera, Wijepurage Sandhya Sulochana |
author_facet | Wickramasinghe, Gammadde Hewa Ishan Maduka Rathnayake, Pilimathalawe Panditharathna Attanayake Mudiyanselage Samith Indika Chandrasekharan, Naduviladath Vishvanath Weerasinghe, Mahindagoda Siril Samantha Wijesundera, Ravindra Lakshman Chundananda Wijesundera, Wijepurage Sandhya Sulochana |
author_sort | Wickramasinghe, Gammadde Hewa Ishan Maduka |
collection | PubMed |
description | It is essential that major carbohydrate polymers in the lignocellulosic biomass are converted into fermentable sugars for the economical production of energy. Xylan, the major component of hemicelluloses, is the second most naturally abundant carbohydrate polymer comprising 20–40% of the total biomass. Endoxylanase (EXN) hydrolyzes xylan into mixtures of xylooligosaccharides. The objective of this study was to genetically modify Pichia stipitis, a pentose sugar fermenting yeast species, to hydrolyze xylan into xylooligosaccharides via cloning and heterologous extracellular expression of EXNI gene from locally isolated Trichoderma virens species. Pichia stipitis was engineered to carry the EXNI gene of T. virens using pGAPZα expression vector. The open reading frame encodes 191 amino acids and SDS-PAGE analysis revealed a 24 kDA recombinant protein. The EXNI activity expressed by recombinant P. stipitis clone under standard conditions using 1% beechwood xylan was 31.7 U/ml. Molecular docking and molecular dynamics simulations were performed to investigate EXNI-xylan interactions. Free EXNI and xylan bound EXNI exhibited similar stabilities and structural behavior in aqueous medium. Furthermore, this in silico work opens avenues for the development of newer generation EXN proteins that can perform better and have enhanced catalytic activity. |
format | Online Article Text |
id | pubmed-5569632 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
spelling | pubmed-55696322017-08-30 Expression, Docking, and Molecular Dynamics of Endo-β-1,4-xylanase I Gene of Trichoderma virens in Pichia stipitis Wickramasinghe, Gammadde Hewa Ishan Maduka Rathnayake, Pilimathalawe Panditharathna Attanayake Mudiyanselage Samith Indika Chandrasekharan, Naduviladath Vishvanath Weerasinghe, Mahindagoda Siril Samantha Wijesundera, Ravindra Lakshman Chundananda Wijesundera, Wijepurage Sandhya Sulochana Biomed Res Int Research Article It is essential that major carbohydrate polymers in the lignocellulosic biomass are converted into fermentable sugars for the economical production of energy. Xylan, the major component of hemicelluloses, is the second most naturally abundant carbohydrate polymer comprising 20–40% of the total biomass. Endoxylanase (EXN) hydrolyzes xylan into mixtures of xylooligosaccharides. The objective of this study was to genetically modify Pichia stipitis, a pentose sugar fermenting yeast species, to hydrolyze xylan into xylooligosaccharides via cloning and heterologous extracellular expression of EXNI gene from locally isolated Trichoderma virens species. Pichia stipitis was engineered to carry the EXNI gene of T. virens using pGAPZα expression vector. The open reading frame encodes 191 amino acids and SDS-PAGE analysis revealed a 24 kDA recombinant protein. The EXNI activity expressed by recombinant P. stipitis clone under standard conditions using 1% beechwood xylan was 31.7 U/ml. Molecular docking and molecular dynamics simulations were performed to investigate EXNI-xylan interactions. Free EXNI and xylan bound EXNI exhibited similar stabilities and structural behavior in aqueous medium. Furthermore, this in silico work opens avenues for the development of newer generation EXN proteins that can perform better and have enhanced catalytic activity. Hindawi 2017 2017-08-10 /pmc/articles/PMC5569632/ /pubmed/28856159 http://dx.doi.org/10.1155/2017/4658584 Text en Copyright © 2017 Gammadde Hewa Ishan Maduka Wickramasinghe et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Wickramasinghe, Gammadde Hewa Ishan Maduka Rathnayake, Pilimathalawe Panditharathna Attanayake Mudiyanselage Samith Indika Chandrasekharan, Naduviladath Vishvanath Weerasinghe, Mahindagoda Siril Samantha Wijesundera, Ravindra Lakshman Chundananda Wijesundera, Wijepurage Sandhya Sulochana Expression, Docking, and Molecular Dynamics of Endo-β-1,4-xylanase I Gene of Trichoderma virens in Pichia stipitis |
title | Expression, Docking, and Molecular Dynamics of Endo-β-1,4-xylanase I Gene of Trichoderma virens in Pichia stipitis |
title_full | Expression, Docking, and Molecular Dynamics of Endo-β-1,4-xylanase I Gene of Trichoderma virens in Pichia stipitis |
title_fullStr | Expression, Docking, and Molecular Dynamics of Endo-β-1,4-xylanase I Gene of Trichoderma virens in Pichia stipitis |
title_full_unstemmed | Expression, Docking, and Molecular Dynamics of Endo-β-1,4-xylanase I Gene of Trichoderma virens in Pichia stipitis |
title_short | Expression, Docking, and Molecular Dynamics of Endo-β-1,4-xylanase I Gene of Trichoderma virens in Pichia stipitis |
title_sort | expression, docking, and molecular dynamics of endo-β-1,4-xylanase i gene of trichoderma virens in pichia stipitis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5569632/ https://www.ncbi.nlm.nih.gov/pubmed/28856159 http://dx.doi.org/10.1155/2017/4658584 |
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