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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2017
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.
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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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