Cargando…

Condition-specific promoter activities in Saccharomyces cerevisiae

BACKGROUND: Saccharomyces cerevisiae is widely studied for production of biofuels and biochemicals. To improve production efficiency under industrially relevant conditions, coordinated expression of multiple genes by manipulating promoter strengths is an efficient approach. It is known that gene exp...

Descripción completa

Detalles Bibliográficos
Autores principales: Xiong, Liang, Zeng, Yu, Tang, Rui-Qi, Alper, Hal S., Bai, Feng-Wu, Zhao, Xin-Qing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5891911/
https://www.ncbi.nlm.nih.gov/pubmed/29631591
http://dx.doi.org/10.1186/s12934-018-0899-6
_version_ 1783313073849761792
author Xiong, Liang
Zeng, Yu
Tang, Rui-Qi
Alper, Hal S.
Bai, Feng-Wu
Zhao, Xin-Qing
author_facet Xiong, Liang
Zeng, Yu
Tang, Rui-Qi
Alper, Hal S.
Bai, Feng-Wu
Zhao, Xin-Qing
author_sort Xiong, Liang
collection PubMed
description BACKGROUND: Saccharomyces cerevisiae is widely studied for production of biofuels and biochemicals. To improve production efficiency under industrially relevant conditions, coordinated expression of multiple genes by manipulating promoter strengths is an efficient approach. It is known that gene expression is highly dependent on the practically used environmental conditions and is subject to dynamic changes. Therefore, investigating promoter activities of S. cerevisiae under different culture conditions in different time points, especially under stressful conditions is of great importance. RESULTS: In this study, the activities of various promoters in S. cerevisiae under stressful conditions and in the presence of xylose were characterized using yeast enhanced green fluorescent protein (yEGFP) as a reporter. The stresses include toxic levels of acetic acid and furfural, and high temperature, which are related to fermentation of lignocellulosic hydrolysates. In addition to investigating eight native promoters, the synthetic hybrid promoter P(3xC-TEF1) was also evaluated. The results revealed that P(TDH3) and the synthetic promoter P(3xC-TEF1) showed the highest strengths under almost all the conditions. Importantly, these two promoters also exhibited high stabilities throughout the cultivation. However, the strengths of P(ADH1) and P(PGK1), which are generally regarded as ‘constitutive’ promoters, decreased significantly under certain conditions, suggesting that cautions should be taken to use such constitutive promoters to drive gene expression under stressful conditions. Interestingly, P(HSP12) and P(HSP26) were able to response to both high temperature and acetic acid stress. Moreover, P(HSP12) also led to moderate yEGFP expression when xylose was used as the sole carbon source, indicating that this promoter could be used for inducing proper gene expression for xylose utilization. CONCLUSION: The results here revealed dynamic changes of promoter activities in S. cerevisiae throughout batch fermentation in the presence of inhibitors as well as using xylose. These results provide insights in selection of promoters to construct S. cerevisiae strains for efficient bioproduction under practical conditions. Our results also encouraged applications of synthetic promoters with high stability for yeast strain development. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0899-6) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-5891911
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-58919112018-04-11 Condition-specific promoter activities in Saccharomyces cerevisiae Xiong, Liang Zeng, Yu Tang, Rui-Qi Alper, Hal S. Bai, Feng-Wu Zhao, Xin-Qing Microb Cell Fact Research BACKGROUND: Saccharomyces cerevisiae is widely studied for production of biofuels and biochemicals. To improve production efficiency under industrially relevant conditions, coordinated expression of multiple genes by manipulating promoter strengths is an efficient approach. It is known that gene expression is highly dependent on the practically used environmental conditions and is subject to dynamic changes. Therefore, investigating promoter activities of S. cerevisiae under different culture conditions in different time points, especially under stressful conditions is of great importance. RESULTS: In this study, the activities of various promoters in S. cerevisiae under stressful conditions and in the presence of xylose were characterized using yeast enhanced green fluorescent protein (yEGFP) as a reporter. The stresses include toxic levels of acetic acid and furfural, and high temperature, which are related to fermentation of lignocellulosic hydrolysates. In addition to investigating eight native promoters, the synthetic hybrid promoter P(3xC-TEF1) was also evaluated. The results revealed that P(TDH3) and the synthetic promoter P(3xC-TEF1) showed the highest strengths under almost all the conditions. Importantly, these two promoters also exhibited high stabilities throughout the cultivation. However, the strengths of P(ADH1) and P(PGK1), which are generally regarded as ‘constitutive’ promoters, decreased significantly under certain conditions, suggesting that cautions should be taken to use such constitutive promoters to drive gene expression under stressful conditions. Interestingly, P(HSP12) and P(HSP26) were able to response to both high temperature and acetic acid stress. Moreover, P(HSP12) also led to moderate yEGFP expression when xylose was used as the sole carbon source, indicating that this promoter could be used for inducing proper gene expression for xylose utilization. CONCLUSION: The results here revealed dynamic changes of promoter activities in S. cerevisiae throughout batch fermentation in the presence of inhibitors as well as using xylose. These results provide insights in selection of promoters to construct S. cerevisiae strains for efficient bioproduction under practical conditions. Our results also encouraged applications of synthetic promoters with high stability for yeast strain development. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0899-6) contains supplementary material, which is available to authorized users. BioMed Central 2018-04-10 /pmc/articles/PMC5891911/ /pubmed/29631591 http://dx.doi.org/10.1186/s12934-018-0899-6 Text en © The Author(s) 2018 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
Xiong, Liang
Zeng, Yu
Tang, Rui-Qi
Alper, Hal S.
Bai, Feng-Wu
Zhao, Xin-Qing
Condition-specific promoter activities in Saccharomyces cerevisiae
title Condition-specific promoter activities in Saccharomyces cerevisiae
title_full Condition-specific promoter activities in Saccharomyces cerevisiae
title_fullStr Condition-specific promoter activities in Saccharomyces cerevisiae
title_full_unstemmed Condition-specific promoter activities in Saccharomyces cerevisiae
title_short Condition-specific promoter activities in Saccharomyces cerevisiae
title_sort condition-specific promoter activities in saccharomyces cerevisiae
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5891911/
https://www.ncbi.nlm.nih.gov/pubmed/29631591
http://dx.doi.org/10.1186/s12934-018-0899-6
work_keys_str_mv AT xiongliang conditionspecificpromoteractivitiesinsaccharomycescerevisiae
AT zengyu conditionspecificpromoteractivitiesinsaccharomycescerevisiae
AT tangruiqi conditionspecificpromoteractivitiesinsaccharomycescerevisiae
AT alperhals conditionspecificpromoteractivitiesinsaccharomycescerevisiae
AT baifengwu conditionspecificpromoteractivitiesinsaccharomycescerevisiae
AT zhaoxinqing conditionspecificpromoteractivitiesinsaccharomycescerevisiae