Cargando…

Engineering a natural Saccharomyces cerevisiae strain for ethanol production from inulin by consolidated bioprocessing

BACKGROUND: The yeast Saccharomyces cerevisiae is an important eukaryotic workhorse in traditional and modern biotechnology. At present, only a few S. cerevisiae strains have been extensively used as engineering hosts. Recently, an astonishing genotypic and phenotypic diversity of S. cerevisiae was...

Descripción completa

Detalles Bibliográficos
Autores principales: Wang, Da, Li, Fu-Li, Wang, Shi-An
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4851821/
https://www.ncbi.nlm.nih.gov/pubmed/27134653
http://dx.doi.org/10.1186/s13068-016-0511-4
_version_ 1782429869353730048
author Wang, Da
Li, Fu-Li
Wang, Shi-An
author_facet Wang, Da
Li, Fu-Li
Wang, Shi-An
author_sort Wang, Da
collection PubMed
description BACKGROUND: The yeast Saccharomyces cerevisiae is an important eukaryotic workhorse in traditional and modern biotechnology. At present, only a few S. cerevisiae strains have been extensively used as engineering hosts. Recently, an astonishing genotypic and phenotypic diversity of S. cerevisiae was disclosed in natural populations. We suppose that some natural strains can be recruited as superior host candidates in bioengineering. This study engineered a natural S. cerevisiae strain with advantages in inulin utilization to produce ethanol from inulin resources by consolidated bioprocess. Rational engineering strategies were employed, including secretive co-expression of heterologous exo- and endo-inulinases, repression of a protease, and switch between haploid and diploid strains. RESULTS: Results from co-expressing endo- and exo-inulinase genes showed that the extracellular inulinase activity increased 20 to 30-fold in engineered S. cerevisiae strains. Repression of the protease PEP4 influenced cell physiology in late stationary phase. Comparison between haploid and diploid engineered strains indicated that diploid strains were superior to haploid strains in ethanol production albeit not in production and secretion of inulinases. Ethanol fermentation from both inulin and Jerusalem artichoke tuber powder was dramatically improved in most engineered strains. Ethanol yield achieved in the ultimate diploid strain JZD-InuMKCP was close to the theoretical maximum. Productivity achieved in the strain JZD-InuMKCP reached to 2.44 and 3.13 g/L/h in fermentation from 200 g/L inulin and 250 g/L raw Jerusalem artichoke tuber powder, respectively. To our knowledge, these are the highest productivities reported up to now in ethanol fermentation from inulin resources. CONCLUSIONS: Although model S. cerevisiae strains are preferentially used as hosts in bioengineering, some natural strains do have specific excellent properties. This study successfully engineered a natural S. cerevisiae strain for efficient ethanol production from inulin resources by consolidated bioprocess, which indicated the feasibility of natural strains used as bioengineering hosts. This study also presented different properties in enzyme secretion and ethanol fermentation between haploid and diploid engineering strains. These findings provided guidelines for host selection in bioengineering. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0511-4) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4851821
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-48518212016-05-01 Engineering a natural Saccharomyces cerevisiae strain for ethanol production from inulin by consolidated bioprocessing Wang, Da Li, Fu-Li Wang, Shi-An Biotechnol Biofuels Research BACKGROUND: The yeast Saccharomyces cerevisiae is an important eukaryotic workhorse in traditional and modern biotechnology. At present, only a few S. cerevisiae strains have been extensively used as engineering hosts. Recently, an astonishing genotypic and phenotypic diversity of S. cerevisiae was disclosed in natural populations. We suppose that some natural strains can be recruited as superior host candidates in bioengineering. This study engineered a natural S. cerevisiae strain with advantages in inulin utilization to produce ethanol from inulin resources by consolidated bioprocess. Rational engineering strategies were employed, including secretive co-expression of heterologous exo- and endo-inulinases, repression of a protease, and switch between haploid and diploid strains. RESULTS: Results from co-expressing endo- and exo-inulinase genes showed that the extracellular inulinase activity increased 20 to 30-fold in engineered S. cerevisiae strains. Repression of the protease PEP4 influenced cell physiology in late stationary phase. Comparison between haploid and diploid engineered strains indicated that diploid strains were superior to haploid strains in ethanol production albeit not in production and secretion of inulinases. Ethanol fermentation from both inulin and Jerusalem artichoke tuber powder was dramatically improved in most engineered strains. Ethanol yield achieved in the ultimate diploid strain JZD-InuMKCP was close to the theoretical maximum. Productivity achieved in the strain JZD-InuMKCP reached to 2.44 and 3.13 g/L/h in fermentation from 200 g/L inulin and 250 g/L raw Jerusalem artichoke tuber powder, respectively. To our knowledge, these are the highest productivities reported up to now in ethanol fermentation from inulin resources. CONCLUSIONS: Although model S. cerevisiae strains are preferentially used as hosts in bioengineering, some natural strains do have specific excellent properties. This study successfully engineered a natural S. cerevisiae strain for efficient ethanol production from inulin resources by consolidated bioprocess, which indicated the feasibility of natural strains used as bioengineering hosts. This study also presented different properties in enzyme secretion and ethanol fermentation between haploid and diploid engineering strains. These findings provided guidelines for host selection in bioengineering. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0511-4) contains supplementary material, which is available to authorized users. BioMed Central 2016-04-30 /pmc/articles/PMC4851821/ /pubmed/27134653 http://dx.doi.org/10.1186/s13068-016-0511-4 Text en © Wang et al. 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
Wang, Da
Li, Fu-Li
Wang, Shi-An
Engineering a natural Saccharomyces cerevisiae strain for ethanol production from inulin by consolidated bioprocessing
title Engineering a natural Saccharomyces cerevisiae strain for ethanol production from inulin by consolidated bioprocessing
title_full Engineering a natural Saccharomyces cerevisiae strain for ethanol production from inulin by consolidated bioprocessing
title_fullStr Engineering a natural Saccharomyces cerevisiae strain for ethanol production from inulin by consolidated bioprocessing
title_full_unstemmed Engineering a natural Saccharomyces cerevisiae strain for ethanol production from inulin by consolidated bioprocessing
title_short Engineering a natural Saccharomyces cerevisiae strain for ethanol production from inulin by consolidated bioprocessing
title_sort engineering a natural saccharomyces cerevisiae strain for ethanol production from inulin by consolidated bioprocessing
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4851821/
https://www.ncbi.nlm.nih.gov/pubmed/27134653
http://dx.doi.org/10.1186/s13068-016-0511-4
work_keys_str_mv AT wangda engineeringanaturalsaccharomycescerevisiaestrainforethanolproductionfrominulinbyconsolidatedbioprocessing
AT lifuli engineeringanaturalsaccharomycescerevisiaestrainforethanolproductionfrominulinbyconsolidatedbioprocessing
AT wangshian engineeringanaturalsaccharomycescerevisiaestrainforethanolproductionfrominulinbyconsolidatedbioprocessing