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Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources

BACKGROUND: As a sustainable industrial process, the production of dicarboxylic acids (DCAs), used as precursors of polyamides, polyesters, perfumes, plasticizers, lubricants, and adhesives, from vegetable oil has continuously garnered interest. Although the yeast Candida tropicalis has been used as...

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Autores principales: Lee, Heeseok, Han, Changpyo, Lee, Hyeok-Won, Park, Gyuyeon, Jeon, Wooyoung, Ahn, Jungoh, Lee, Hongweon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6225622/
https://www.ncbi.nlm.nih.gov/pubmed/30455739
http://dx.doi.org/10.1186/s13068-018-1310-x
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author Lee, Heeseok
Han, Changpyo
Lee, Hyeok-Won
Park, Gyuyeon
Jeon, Wooyoung
Ahn, Jungoh
Lee, Hongweon
author_facet Lee, Heeseok
Han, Changpyo
Lee, Hyeok-Won
Park, Gyuyeon
Jeon, Wooyoung
Ahn, Jungoh
Lee, Hongweon
author_sort Lee, Heeseok
collection PubMed
description BACKGROUND: As a sustainable industrial process, the production of dicarboxylic acids (DCAs), used as precursors of polyamides, polyesters, perfumes, plasticizers, lubricants, and adhesives, from vegetable oil has continuously garnered interest. Although the yeast Candida tropicalis has been used as a host for DCA production, additional strains are continually investigated to meet productivity thresholds and industrial needs. In this regard, the yeast Wickerhamiella sorbophila, a potential candidate strain, has been screened. However, the lack of genetic and physiological information for this uncommon strain is an obstacle that merits further research. To overcome this limitation, we attempted to develop a method to facilitate genetic recombination in this strain and produce high amounts of DCAs from methyl laurate using engineered W. sorbophila. RESULTS: In the current study, we first developed efficient genetic engineering tools for the industrial application of W. sorbophila. To increase homologous recombination (HR) efficiency during transformation, the cell cycle of the yeast was synchronized to the S/G2 phase using hydroxyurea. The HR efficiency at POX1 and POX2 loci increased from 56.3% and 41.7%, respectively, to 97.9% in both cases. The original HR efficiency at URA3 and ADE2 loci was nearly 0% during the early stationary and logarithmic phases of growth, and increased to 4.8% and 25.6%, respectively. We used the developed tools to construct W. sorbophila UHP4, in which β-oxidation was completely blocked. The strain produced 92.5 g/l of dodecanedioic acid (DDDA) from methyl laurate over 126 h in 5-l fed-batch fermentation, with a productivity of 0.83 g/l/h. CONCLUSIONS: Wickerhamiella sorbophila UHP4 produced more DDDA methyl laurate than C. tropicalis. Hence, we demonstrated that W. sorbophila is a powerful microbial platform for vegetable oil-based DCA production. In addition, by using the developed genetic engineering tools, this emerging yeast could be used for the production of a variety of fatty acid derivatives, such as fatty alcohols, fatty aldehydes, and ω-hydroxy fatty acids. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-018-1310-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-62256222018-11-19 Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources Lee, Heeseok Han, Changpyo Lee, Hyeok-Won Park, Gyuyeon Jeon, Wooyoung Ahn, Jungoh Lee, Hongweon Biotechnol Biofuels Research BACKGROUND: As a sustainable industrial process, the production of dicarboxylic acids (DCAs), used as precursors of polyamides, polyesters, perfumes, plasticizers, lubricants, and adhesives, from vegetable oil has continuously garnered interest. Although the yeast Candida tropicalis has been used as a host for DCA production, additional strains are continually investigated to meet productivity thresholds and industrial needs. In this regard, the yeast Wickerhamiella sorbophila, a potential candidate strain, has been screened. However, the lack of genetic and physiological information for this uncommon strain is an obstacle that merits further research. To overcome this limitation, we attempted to develop a method to facilitate genetic recombination in this strain and produce high amounts of DCAs from methyl laurate using engineered W. sorbophila. RESULTS: In the current study, we first developed efficient genetic engineering tools for the industrial application of W. sorbophila. To increase homologous recombination (HR) efficiency during transformation, the cell cycle of the yeast was synchronized to the S/G2 phase using hydroxyurea. The HR efficiency at POX1 and POX2 loci increased from 56.3% and 41.7%, respectively, to 97.9% in both cases. The original HR efficiency at URA3 and ADE2 loci was nearly 0% during the early stationary and logarithmic phases of growth, and increased to 4.8% and 25.6%, respectively. We used the developed tools to construct W. sorbophila UHP4, in which β-oxidation was completely blocked. The strain produced 92.5 g/l of dodecanedioic acid (DDDA) from methyl laurate over 126 h in 5-l fed-batch fermentation, with a productivity of 0.83 g/l/h. CONCLUSIONS: Wickerhamiella sorbophila UHP4 produced more DDDA methyl laurate than C. tropicalis. Hence, we demonstrated that W. sorbophila is a powerful microbial platform for vegetable oil-based DCA production. In addition, by using the developed genetic engineering tools, this emerging yeast could be used for the production of a variety of fatty acid derivatives, such as fatty alcohols, fatty aldehydes, and ω-hydroxy fatty acids. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-018-1310-x) contains supplementary material, which is available to authorized users. BioMed Central 2018-11-09 /pmc/articles/PMC6225622/ /pubmed/30455739 http://dx.doi.org/10.1186/s13068-018-1310-x 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
Lee, Heeseok
Han, Changpyo
Lee, Hyeok-Won
Park, Gyuyeon
Jeon, Wooyoung
Ahn, Jungoh
Lee, Hongweon
Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources
title Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources
title_full Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources
title_fullStr Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources
title_full_unstemmed Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources
title_short Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources
title_sort development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6225622/
https://www.ncbi.nlm.nih.gov/pubmed/30455739
http://dx.doi.org/10.1186/s13068-018-1310-x
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