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Proof of concept for the simplified breakdown of cellulose by combining Pseudomonasputida strains with surface displayed thermophilic endocellulase, exocellulase and β-glucosidase
BACKGROUND: The production and employment of cellulases still represents an economic bottleneck in the conversion of lignocellulosic biomass to biofuels and other biocommodities. This process could be simplified by displaying the necessary enzymes on a microbial cell surface. Such an approach, howev...
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/PMC4901517/ https://www.ncbi.nlm.nih.gov/pubmed/27287198 http://dx.doi.org/10.1186/s12934-016-0505-8 |
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author | Tozakidis, Iasson E. P. Brossette, Tatjana Lenz, Florian Maas, Ruth M. Jose, Joachim |
author_facet | Tozakidis, Iasson E. P. Brossette, Tatjana Lenz, Florian Maas, Ruth M. Jose, Joachim |
author_sort | Tozakidis, Iasson E. P. |
collection | PubMed |
description | BACKGROUND: The production and employment of cellulases still represents an economic bottleneck in the conversion of lignocellulosic biomass to biofuels and other biocommodities. This process could be simplified by displaying the necessary enzymes on a microbial cell surface. Such an approach, however, requires an appropriate host organism which on the one hand can withstand the rough environment coming along with lignocellulose hydrolysis, and on the other hand does not consume the generated glucose so that it remains available for subsequent fermentation steps. RESULTS: The robust soil bacterium Pseudomonas putida showed a strongly reduced uptake of glucose above a temperature of 50 °C, while remaining structurally intact hence recyclable, which makes it suitable for cellulose hydrolysis at elevated temperatures. Consequently, three complementary, thermophilic cellulases from Ruminiclostridium thermocellum were displayed on the surface of the bacterium. All three enzymes retained their activity on the cell surface. A mixture of three strains displaying each one of these enzymes was able to synergistically hydrolyze filter paper at 55 °C, producing 20 μg glucose per mL cell suspension in 24 h. CONCLUSION: We could establish Pseudomonas putida as host for the surface display of cellulases, and provided proof-of-concept for a fast and simple cellulose breakdown process at elevated temperatures. This study opens up new perspectives for the application of P. putida in the production of biofuels and other biotechnological products. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-016-0505-8) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4901517 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-49015172016-06-11 Proof of concept for the simplified breakdown of cellulose by combining Pseudomonasputida strains with surface displayed thermophilic endocellulase, exocellulase and β-glucosidase Tozakidis, Iasson E. P. Brossette, Tatjana Lenz, Florian Maas, Ruth M. Jose, Joachim Microb Cell Fact Research BACKGROUND: The production and employment of cellulases still represents an economic bottleneck in the conversion of lignocellulosic biomass to biofuels and other biocommodities. This process could be simplified by displaying the necessary enzymes on a microbial cell surface. Such an approach, however, requires an appropriate host organism which on the one hand can withstand the rough environment coming along with lignocellulose hydrolysis, and on the other hand does not consume the generated glucose so that it remains available for subsequent fermentation steps. RESULTS: The robust soil bacterium Pseudomonas putida showed a strongly reduced uptake of glucose above a temperature of 50 °C, while remaining structurally intact hence recyclable, which makes it suitable for cellulose hydrolysis at elevated temperatures. Consequently, three complementary, thermophilic cellulases from Ruminiclostridium thermocellum were displayed on the surface of the bacterium. All three enzymes retained their activity on the cell surface. A mixture of three strains displaying each one of these enzymes was able to synergistically hydrolyze filter paper at 55 °C, producing 20 μg glucose per mL cell suspension in 24 h. CONCLUSION: We could establish Pseudomonas putida as host for the surface display of cellulases, and provided proof-of-concept for a fast and simple cellulose breakdown process at elevated temperatures. This study opens up new perspectives for the application of P. putida in the production of biofuels and other biotechnological products. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-016-0505-8) contains supplementary material, which is available to authorized users. BioMed Central 2016-06-10 /pmc/articles/PMC4901517/ /pubmed/27287198 http://dx.doi.org/10.1186/s12934-016-0505-8 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 Tozakidis, Iasson E. P. Brossette, Tatjana Lenz, Florian Maas, Ruth M. Jose, Joachim Proof of concept for the simplified breakdown of cellulose by combining Pseudomonasputida strains with surface displayed thermophilic endocellulase, exocellulase and β-glucosidase |
title | Proof of concept for the simplified breakdown of cellulose by combining Pseudomonasputida strains with surface displayed thermophilic endocellulase, exocellulase and β-glucosidase |
title_full | Proof of concept for the simplified breakdown of cellulose by combining Pseudomonasputida strains with surface displayed thermophilic endocellulase, exocellulase and β-glucosidase |
title_fullStr | Proof of concept for the simplified breakdown of cellulose by combining Pseudomonasputida strains with surface displayed thermophilic endocellulase, exocellulase and β-glucosidase |
title_full_unstemmed | Proof of concept for the simplified breakdown of cellulose by combining Pseudomonasputida strains with surface displayed thermophilic endocellulase, exocellulase and β-glucosidase |
title_short | Proof of concept for the simplified breakdown of cellulose by combining Pseudomonasputida strains with surface displayed thermophilic endocellulase, exocellulase and β-glucosidase |
title_sort | proof of concept for the simplified breakdown of cellulose by combining pseudomonasputida strains with surface displayed thermophilic endocellulase, exocellulase and β-glucosidase |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4901517/ https://www.ncbi.nlm.nih.gov/pubmed/27287198 http://dx.doi.org/10.1186/s12934-016-0505-8 |
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