Cargando…

A highly efficient β-glucosidase from the buffalo rumen fungus Neocallimastix patriciarum W5

BACKGROUND: Cellulose, which is the most abundant renewable biomass on earth, is a potential bio-resource of alternative energy. The hydrolysis of plant polysaccharides is catalyzed by microbial cellulases, including endo-β-1,4-glucanases, cellobiohydrolases, cellodextrinases, and β-glucosidases. Co...

Descripción completa

Detalles Bibliográficos
Autores principales: Chen, Hsin-Liang, Chen, Yo-Chia, Lu, Mei-Yeh Jade, Chang, Jui-Jen, Wang, Hiaow-Ting Christine, Ke, Huei-Mien, Wang, Tzi-Yuan, Ruan, Sz-Kai, Wang, Tao-Yuan, Hung, Kuo-Yen, Cho, Hsing-Yi, Lin, Wan-Ting, Shih, Ming-Che, Li, Wen-Hsiung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3403894/
https://www.ncbi.nlm.nih.gov/pubmed/22515264
http://dx.doi.org/10.1186/1754-6834-5-24
_version_ 1782238939490287616
author Chen, Hsin-Liang
Chen, Yo-Chia
Lu, Mei-Yeh Jade
Chang, Jui-Jen
Wang, Hiaow-Ting Christine
Ke, Huei-Mien
Wang, Tzi-Yuan
Ruan, Sz-Kai
Wang, Tao-Yuan
Hung, Kuo-Yen
Cho, Hsing-Yi
Lin, Wan-Ting
Shih, Ming-Che
Li, Wen-Hsiung
author_facet Chen, Hsin-Liang
Chen, Yo-Chia
Lu, Mei-Yeh Jade
Chang, Jui-Jen
Wang, Hiaow-Ting Christine
Ke, Huei-Mien
Wang, Tzi-Yuan
Ruan, Sz-Kai
Wang, Tao-Yuan
Hung, Kuo-Yen
Cho, Hsing-Yi
Lin, Wan-Ting
Shih, Ming-Che
Li, Wen-Hsiung
author_sort Chen, Hsin-Liang
collection PubMed
description BACKGROUND: Cellulose, which is the most abundant renewable biomass on earth, is a potential bio-resource of alternative energy. The hydrolysis of plant polysaccharides is catalyzed by microbial cellulases, including endo-β-1,4-glucanases, cellobiohydrolases, cellodextrinases, and β-glucosidases. Converting cellobiose by β-glucosidases is the key factor for reducing cellobiose inhibition and enhancing the efficiency of cellulolytic enzymes for cellulosic ethanol production. RESULTS: In this study, a cDNA encoding β-glucosidase was isolated from the buffalo rumen fungus Neocallimastix patriciarum W5 and is named NpaBGS. It has a length of 2,331 bp with an open reading frame coding for a protein of 776 amino acid residues, corresponding to a theoretical molecular mass of 85.1 kDa and isoelectric point of 4.4. Two GH3 catalytic domains were found at the N and C terminals of NpaBGS by sequence analysis. The cDNA was expressed in Pichia pastoris and after protein purification, the enzyme displayed a specific activity of 34.5 U/mg against cellobiose as the substrate. Enzymatic assays showed that NpaBGS was active on short cello-oligosaccharides from various substrates. A weak activity in carboxymethyl cellulose (CMC) digestion indicated that the enzyme might also have the function of an endoglucanase. The optimal activity was detected at 40°C and pH 5 ~ 6, showing that the enzyme prefers a weak acid condition. Moreover, its activity could be enhanced at 50°C by adding Mg(2+) or Mn(2+) ions. Interestingly, in simultaneous saccharification and fermentation (SSF) experiments using Saccharomyces cerevisiae BY4741 or Kluyveromyces marxianus KY3 as the fermentation yeast, NpaBGS showed advantages in cell growth, glucose production, and ethanol production over the commercial enzyme Novo 188. Moreover, we showed that the KY3 strain engineered with the NpaNGS gene can utilize 2 % dry napiergrass as the sole carbon source to produce 3.32 mg/ml ethanol when Celluclast 1.5 L was added to the SSF system. CONCLUSION: Our characterizations of the novel β-glucosidase NpaBGS revealed that it has a preference of weak acidity for optimal yeast fermentation and an optimal temperature of ~40°C. Since NpaBGS performs better than Novo 188 under the living conditions of fermentation yeasts, it has the potential to be a suitable enzyme for SSF.
format Online
Article
Text
id pubmed-3403894
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-34038942012-07-25 A highly efficient β-glucosidase from the buffalo rumen fungus Neocallimastix patriciarum W5 Chen, Hsin-Liang Chen, Yo-Chia Lu, Mei-Yeh Jade Chang, Jui-Jen Wang, Hiaow-Ting Christine Ke, Huei-Mien Wang, Tzi-Yuan Ruan, Sz-Kai Wang, Tao-Yuan Hung, Kuo-Yen Cho, Hsing-Yi Lin, Wan-Ting Shih, Ming-Che Li, Wen-Hsiung Biotechnol Biofuels Research BACKGROUND: Cellulose, which is the most abundant renewable biomass on earth, is a potential bio-resource of alternative energy. The hydrolysis of plant polysaccharides is catalyzed by microbial cellulases, including endo-β-1,4-glucanases, cellobiohydrolases, cellodextrinases, and β-glucosidases. Converting cellobiose by β-glucosidases is the key factor for reducing cellobiose inhibition and enhancing the efficiency of cellulolytic enzymes for cellulosic ethanol production. RESULTS: In this study, a cDNA encoding β-glucosidase was isolated from the buffalo rumen fungus Neocallimastix patriciarum W5 and is named NpaBGS. It has a length of 2,331 bp with an open reading frame coding for a protein of 776 amino acid residues, corresponding to a theoretical molecular mass of 85.1 kDa and isoelectric point of 4.4. Two GH3 catalytic domains were found at the N and C terminals of NpaBGS by sequence analysis. The cDNA was expressed in Pichia pastoris and after protein purification, the enzyme displayed a specific activity of 34.5 U/mg against cellobiose as the substrate. Enzymatic assays showed that NpaBGS was active on short cello-oligosaccharides from various substrates. A weak activity in carboxymethyl cellulose (CMC) digestion indicated that the enzyme might also have the function of an endoglucanase. The optimal activity was detected at 40°C and pH 5 ~ 6, showing that the enzyme prefers a weak acid condition. Moreover, its activity could be enhanced at 50°C by adding Mg(2+) or Mn(2+) ions. Interestingly, in simultaneous saccharification and fermentation (SSF) experiments using Saccharomyces cerevisiae BY4741 or Kluyveromyces marxianus KY3 as the fermentation yeast, NpaBGS showed advantages in cell growth, glucose production, and ethanol production over the commercial enzyme Novo 188. Moreover, we showed that the KY3 strain engineered with the NpaNGS gene can utilize 2 % dry napiergrass as the sole carbon source to produce 3.32 mg/ml ethanol when Celluclast 1.5 L was added to the SSF system. CONCLUSION: Our characterizations of the novel β-glucosidase NpaBGS revealed that it has a preference of weak acidity for optimal yeast fermentation and an optimal temperature of ~40°C. Since NpaBGS performs better than Novo 188 under the living conditions of fermentation yeasts, it has the potential to be a suitable enzyme for SSF. BioMed Central 2012-04-19 /pmc/articles/PMC3403894/ /pubmed/22515264 http://dx.doi.org/10.1186/1754-6834-5-24 Text en Copyright © 2012 Chen et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Chen, Hsin-Liang
Chen, Yo-Chia
Lu, Mei-Yeh Jade
Chang, Jui-Jen
Wang, Hiaow-Ting Christine
Ke, Huei-Mien
Wang, Tzi-Yuan
Ruan, Sz-Kai
Wang, Tao-Yuan
Hung, Kuo-Yen
Cho, Hsing-Yi
Lin, Wan-Ting
Shih, Ming-Che
Li, Wen-Hsiung
A highly efficient β-glucosidase from the buffalo rumen fungus Neocallimastix patriciarum W5
title A highly efficient β-glucosidase from the buffalo rumen fungus Neocallimastix patriciarum W5
title_full A highly efficient β-glucosidase from the buffalo rumen fungus Neocallimastix patriciarum W5
title_fullStr A highly efficient β-glucosidase from the buffalo rumen fungus Neocallimastix patriciarum W5
title_full_unstemmed A highly efficient β-glucosidase from the buffalo rumen fungus Neocallimastix patriciarum W5
title_short A highly efficient β-glucosidase from the buffalo rumen fungus Neocallimastix patriciarum W5
title_sort highly efficient β-glucosidase from the buffalo rumen fungus neocallimastix patriciarum w5
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3403894/
https://www.ncbi.nlm.nih.gov/pubmed/22515264
http://dx.doi.org/10.1186/1754-6834-5-24
work_keys_str_mv AT chenhsinliang ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT chenyochia ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT lumeiyehjade ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT changjuijen ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT wanghiaowtingchristine ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT kehueimien ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT wangtziyuan ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT ruanszkai ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT wangtaoyuan ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT hungkuoyen ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT chohsingyi ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT linwanting ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT shihmingche ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT liwenhsiung ahighlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT chenhsinliang highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT chenyochia highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT lumeiyehjade highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT changjuijen highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT wanghiaowtingchristine highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT kehueimien highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT wangtziyuan highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT ruanszkai highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT wangtaoyuan highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT hungkuoyen highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT chohsingyi highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT linwanting highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT shihmingche highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5
AT liwenhsiung highlyefficientbglucosidasefromthebuffalorumenfungusneocallimastixpatriciarumw5