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From mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast Saccharomyces cerevisiae
BACKGROUND: Mannans represent the largest hemicellulosic fraction in softwoods and also serve as carbohydrate stores in various plants. However, the utilization of mannans as sustainable resources has been less advanced in sustainable biofuel development. Based on a yeast cell surface-display techno...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5009545/ https://www.ncbi.nlm.nih.gov/pubmed/27594915 http://dx.doi.org/10.1186/s13068-016-0600-4 |
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author | Ishii, Jun Okazaki, Fumiyoshi Djohan, Apridah Cameliawati Hara, Kiyotaka Y. Asai-Nakashima, Nanami Teramura, Hiroshi Andriani, Ade Tominaga, Masahiro Wakai, Satoshi Kahar, Prihardi Yopi Prasetya, Bambang Ogino, Chiaki Kondo, Akihiko |
author_facet | Ishii, Jun Okazaki, Fumiyoshi Djohan, Apridah Cameliawati Hara, Kiyotaka Y. Asai-Nakashima, Nanami Teramura, Hiroshi Andriani, Ade Tominaga, Masahiro Wakai, Satoshi Kahar, Prihardi Yopi Prasetya, Bambang Ogino, Chiaki Kondo, Akihiko |
author_sort | Ishii, Jun |
collection | PubMed |
description | BACKGROUND: Mannans represent the largest hemicellulosic fraction in softwoods and also serve as carbohydrate stores in various plants. However, the utilization of mannans as sustainable resources has been less advanced in sustainable biofuel development. Based on a yeast cell surface-display technology that enables the immobilization of multiple enzymes on the yeast cell walls, we constructed a recombinant Saccharomyces cerevisiae strain that co-displays β-mannanase and β-mannosidase; this strain is expected to facilitate ethanol fermentation using mannan as a biomass source. RESULTS: Parental yeast S. cerevisiae assimilated mannose and glucose as monomeric sugars, producing ethanol from mannose. We constructed yeast strains that express tethered β-mannanase and β-mannosidase; co-display of the two enzymes on the cell surface was confirmed by immunofluorescence staining and enzyme activity assays. The constructed yeast cells successfully hydrolyzed 1,4-β-d-mannan and produced ethanol by assimilating the resulting mannose without external addition of enzymes. Furthermore, the constructed strain produced ethanol from 1,4-β-d-mannan continually during the third batch of repeated fermentation. Additionally, the constructed strain produced ethanol from ivory nut mannan; ethanol yield was improved by NaOH pretreatment of the substrate. CONCLUSIONS: We successfully displayed β-mannanase and β-mannosidase on the yeast cell surface. Our results clearly demonstrate the utility of the strain co-displaying β-mannanase and β-mannosidase for ethanol fermentation from mannan biomass. Thus, co-tethering β-mannanase and β-mannosidase on the yeast cell surface provides a powerful platform technology for yeast fermentation toward the production of bioethanol and other biochemicals from lignocellulosic materials containing mannan components. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0600-4) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5009545 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-50095452016-09-03 From mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast Saccharomyces cerevisiae Ishii, Jun Okazaki, Fumiyoshi Djohan, Apridah Cameliawati Hara, Kiyotaka Y. Asai-Nakashima, Nanami Teramura, Hiroshi Andriani, Ade Tominaga, Masahiro Wakai, Satoshi Kahar, Prihardi Yopi Prasetya, Bambang Ogino, Chiaki Kondo, Akihiko Biotechnol Biofuels Research BACKGROUND: Mannans represent the largest hemicellulosic fraction in softwoods and also serve as carbohydrate stores in various plants. However, the utilization of mannans as sustainable resources has been less advanced in sustainable biofuel development. Based on a yeast cell surface-display technology that enables the immobilization of multiple enzymes on the yeast cell walls, we constructed a recombinant Saccharomyces cerevisiae strain that co-displays β-mannanase and β-mannosidase; this strain is expected to facilitate ethanol fermentation using mannan as a biomass source. RESULTS: Parental yeast S. cerevisiae assimilated mannose and glucose as monomeric sugars, producing ethanol from mannose. We constructed yeast strains that express tethered β-mannanase and β-mannosidase; co-display of the two enzymes on the cell surface was confirmed by immunofluorescence staining and enzyme activity assays. The constructed yeast cells successfully hydrolyzed 1,4-β-d-mannan and produced ethanol by assimilating the resulting mannose without external addition of enzymes. Furthermore, the constructed strain produced ethanol from 1,4-β-d-mannan continually during the third batch of repeated fermentation. Additionally, the constructed strain produced ethanol from ivory nut mannan; ethanol yield was improved by NaOH pretreatment of the substrate. CONCLUSIONS: We successfully displayed β-mannanase and β-mannosidase on the yeast cell surface. Our results clearly demonstrate the utility of the strain co-displaying β-mannanase and β-mannosidase for ethanol fermentation from mannan biomass. Thus, co-tethering β-mannanase and β-mannosidase on the yeast cell surface provides a powerful platform technology for yeast fermentation toward the production of bioethanol and other biochemicals from lignocellulosic materials containing mannan components. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0600-4) contains supplementary material, which is available to authorized users. BioMed Central 2016-09-02 /pmc/articles/PMC5009545/ /pubmed/27594915 http://dx.doi.org/10.1186/s13068-016-0600-4 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Ishii, Jun Okazaki, Fumiyoshi Djohan, Apridah Cameliawati Hara, Kiyotaka Y. Asai-Nakashima, Nanami Teramura, Hiroshi Andriani, Ade Tominaga, Masahiro Wakai, Satoshi Kahar, Prihardi Yopi Prasetya, Bambang Ogino, Chiaki Kondo, Akihiko From mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast Saccharomyces cerevisiae |
title | From mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast Saccharomyces cerevisiae |
title_full | From mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast Saccharomyces cerevisiae |
title_fullStr | From mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast Saccharomyces cerevisiae |
title_full_unstemmed | From mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast Saccharomyces cerevisiae |
title_short | From mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast Saccharomyces cerevisiae |
title_sort | from mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast saccharomyces cerevisiae |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5009545/ https://www.ncbi.nlm.nih.gov/pubmed/27594915 http://dx.doi.org/10.1186/s13068-016-0600-4 |
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