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Investigation of availability of a high throughput screening method for predicting butanol solvent -producing ability of Clostridium beijerinckii
BACKGROUND: Currently, efficient screening methods for selection of desired bacterial phenotypes from large populations are not easy feasible or readily available due to the complicated physiological and metabolic networks of solventogenic clostridia. In this study, to contribute to the improvement...
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/PMC4957875/ https://www.ncbi.nlm.nih.gov/pubmed/27448996 http://dx.doi.org/10.1186/s12866-016-0776-6 |
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author | Su, HaiFeng Zhu, Jun Liu, Gang Tan, Furong |
author_facet | Su, HaiFeng Zhu, Jun Liu, Gang Tan, Furong |
author_sort | Su, HaiFeng |
collection | PubMed |
description | BACKGROUND: Currently, efficient screening methods for selection of desired bacterial phenotypes from large populations are not easy feasible or readily available due to the complicated physiological and metabolic networks of solventogenic clostridia. In this study, to contribute to the improvement of methods for predicting the butanol-producing ability of Clostridium beijerinckii based on starch substrate, we further investigate a simple, visualization screening method for selecting target strains from mutant library of Clostridium beijerinckii NCIMB 8052 by using trypan blue dye as an indicator in solid starch via statistical survey and validation of fermentation experiment with controlling pH. RESULTS: To verify an effective, efficient phenotypic screening method for isolating high butanol-producing mutants, the revalidation process was conducted based on Trypan Blue was used for visualization, and starch was used as the bacterial metabolic substrate. The availability of the screening system was further evaluated based on the relationship between characteristics of mutant strains and their α-amylase activities. Mutant clones were analyzed in detail based on their distinctive growth patterns and rate of fermentation of soluble starch to form butanol and were compared by statistical method. Significant correlations were identified between colony morphology and changes in butanol concentrations. The screening method was validated via statistical analysis for characterizing phenotypic parameters. The fermentation experiment of mutant strains with controlling pH value also demonstrated a positive correlation between increased α-amylase activity and increased solvent production by Clostridium beijerinckii was observed, and therefore indicated that the trypan blue dyeing method can be used as a fast method to screen target mutant strain for better solvent producers from, for instance, a mutant library. CONCLUSIONS: The suitability of the novel screening procedure was validated, opening up a new indicator of approach to select mutant solventogenic clostridia with improved fermentation of starch to increase butanol concentrations. The applicability can easily be broadened to a wide range of interesting microbes such as cellulolytic or acetogenic microorganisms, which produce biofuels from feedstock rich in starch. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-016-0776-6) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4957875 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-49578752016-07-23 Investigation of availability of a high throughput screening method for predicting butanol solvent -producing ability of Clostridium beijerinckii Su, HaiFeng Zhu, Jun Liu, Gang Tan, Furong BMC Microbiol Methodology Article BACKGROUND: Currently, efficient screening methods for selection of desired bacterial phenotypes from large populations are not easy feasible or readily available due to the complicated physiological and metabolic networks of solventogenic clostridia. In this study, to contribute to the improvement of methods for predicting the butanol-producing ability of Clostridium beijerinckii based on starch substrate, we further investigate a simple, visualization screening method for selecting target strains from mutant library of Clostridium beijerinckii NCIMB 8052 by using trypan blue dye as an indicator in solid starch via statistical survey and validation of fermentation experiment with controlling pH. RESULTS: To verify an effective, efficient phenotypic screening method for isolating high butanol-producing mutants, the revalidation process was conducted based on Trypan Blue was used for visualization, and starch was used as the bacterial metabolic substrate. The availability of the screening system was further evaluated based on the relationship between characteristics of mutant strains and their α-amylase activities. Mutant clones were analyzed in detail based on their distinctive growth patterns and rate of fermentation of soluble starch to form butanol and were compared by statistical method. Significant correlations were identified between colony morphology and changes in butanol concentrations. The screening method was validated via statistical analysis for characterizing phenotypic parameters. The fermentation experiment of mutant strains with controlling pH value also demonstrated a positive correlation between increased α-amylase activity and increased solvent production by Clostridium beijerinckii was observed, and therefore indicated that the trypan blue dyeing method can be used as a fast method to screen target mutant strain for better solvent producers from, for instance, a mutant library. CONCLUSIONS: The suitability of the novel screening procedure was validated, opening up a new indicator of approach to select mutant solventogenic clostridia with improved fermentation of starch to increase butanol concentrations. The applicability can easily be broadened to a wide range of interesting microbes such as cellulolytic or acetogenic microorganisms, which produce biofuels from feedstock rich in starch. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-016-0776-6) contains supplementary material, which is available to authorized users. BioMed Central 2016-07-22 /pmc/articles/PMC4957875/ /pubmed/27448996 http://dx.doi.org/10.1186/s12866-016-0776-6 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 | Methodology Article Su, HaiFeng Zhu, Jun Liu, Gang Tan, Furong Investigation of availability of a high throughput screening method for predicting butanol solvent -producing ability of Clostridium beijerinckii |
title | Investigation of availability of a high throughput screening method for predicting butanol solvent -producing ability of Clostridium beijerinckii |
title_full | Investigation of availability of a high throughput screening method for predicting butanol solvent -producing ability of Clostridium beijerinckii |
title_fullStr | Investigation of availability of a high throughput screening method for predicting butanol solvent -producing ability of Clostridium beijerinckii |
title_full_unstemmed | Investigation of availability of a high throughput screening method for predicting butanol solvent -producing ability of Clostridium beijerinckii |
title_short | Investigation of availability of a high throughput screening method for predicting butanol solvent -producing ability of Clostridium beijerinckii |
title_sort | investigation of availability of a high throughput screening method for predicting butanol solvent -producing ability of clostridium beijerinckii |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4957875/ https://www.ncbi.nlm.nih.gov/pubmed/27448996 http://dx.doi.org/10.1186/s12866-016-0776-6 |
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