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Engineering oleaginous yeast Rhodotorula toruloides for overproduction of fatty acid ethyl esters
BACKGROUND: Production of biofuels and green chemicals by microbes is currently of great interest due to the increasingly limited reserves of fossil fuels. Biodiesel, especially fatty acid ethyl esters (FAEEs), is considered as an attractive alternative because of its similarity with petrodiesel and...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8106135/ https://www.ncbi.nlm.nih.gov/pubmed/33964988 http://dx.doi.org/10.1186/s13068-021-01965-3 |
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author | Zhang, Yang Peng, Jie Zhao, Huimin Shi, Shuobo |
author_facet | Zhang, Yang Peng, Jie Zhao, Huimin Shi, Shuobo |
author_sort | Zhang, Yang |
collection | PubMed |
description | BACKGROUND: Production of biofuels and green chemicals by microbes is currently of great interest due to the increasingly limited reserves of fossil fuels. Biodiesel, especially fatty acid ethyl esters (FAEEs), is considered as an attractive alternative because of its similarity with petrodiesel and compatibility with existing infrastructures. Cost-efficient bio-production of FAEEs requires a highly lipogenic production host that is suitable for large-scale fermentation. As a non-model oleaginous yeast that can be cultured to an extremely high cell density and accumulate over 70% cell mass as lipids, Rhodotorula toruloides represents an attractive host for FAEEs production. RESULTS: We first constructed the FAEE biosynthetic pathways in R. toruloides by introducing various wax ester synthase genes from different sources, and the bifunctional wax ester synthase/acyl-CoA-diacyglycerol acyltransferase (WS/DGAT) gene from Acinetobacter baylyi was successfully expressed, leading to a production of 826 mg/L FAEEs through shake-flask cultivation. We then mutated this bifunctional enzyme to abolish the DGAT activity, and further improved the titer to 1.02 g/L. Finally, to elevate the performance of Δku70-AbWS* in a bioreactor, both batch and fed-batch cultivation strategies were performed. The FAEEs titer, productivity and yield were 4.03 g/L, 69.5 mg/L/h and 57.9 mg/g (mg FAEEs/g glucose) under batch cultivation, and 9.97 g/L, 90.6 mg/L/h, and 86.1 mg/g under fed-batch cultivation. It is worth mentioning that most of the produced FAEEs were secreted out of the cell, which should greatly reduce the cost of downstream processing. CONCLUSION: We achieved the highest FAEEs production in yeast with a final titer of 9.97 g/L and demonstrated that the engineered R. toruloides has the potential to serve as a platform strain for efficient production of fatty acid-derived molecules. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13068-021-01965-3. |
format | Online Article Text |
id | pubmed-8106135 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-81061352021-05-10 Engineering oleaginous yeast Rhodotorula toruloides for overproduction of fatty acid ethyl esters Zhang, Yang Peng, Jie Zhao, Huimin Shi, Shuobo Biotechnol Biofuels Research BACKGROUND: Production of biofuels and green chemicals by microbes is currently of great interest due to the increasingly limited reserves of fossil fuels. Biodiesel, especially fatty acid ethyl esters (FAEEs), is considered as an attractive alternative because of its similarity with petrodiesel and compatibility with existing infrastructures. Cost-efficient bio-production of FAEEs requires a highly lipogenic production host that is suitable for large-scale fermentation. As a non-model oleaginous yeast that can be cultured to an extremely high cell density and accumulate over 70% cell mass as lipids, Rhodotorula toruloides represents an attractive host for FAEEs production. RESULTS: We first constructed the FAEE biosynthetic pathways in R. toruloides by introducing various wax ester synthase genes from different sources, and the bifunctional wax ester synthase/acyl-CoA-diacyglycerol acyltransferase (WS/DGAT) gene from Acinetobacter baylyi was successfully expressed, leading to a production of 826 mg/L FAEEs through shake-flask cultivation. We then mutated this bifunctional enzyme to abolish the DGAT activity, and further improved the titer to 1.02 g/L. Finally, to elevate the performance of Δku70-AbWS* in a bioreactor, both batch and fed-batch cultivation strategies were performed. The FAEEs titer, productivity and yield were 4.03 g/L, 69.5 mg/L/h and 57.9 mg/g (mg FAEEs/g glucose) under batch cultivation, and 9.97 g/L, 90.6 mg/L/h, and 86.1 mg/g under fed-batch cultivation. It is worth mentioning that most of the produced FAEEs were secreted out of the cell, which should greatly reduce the cost of downstream processing. CONCLUSION: We achieved the highest FAEEs production in yeast with a final titer of 9.97 g/L and demonstrated that the engineered R. toruloides has the potential to serve as a platform strain for efficient production of fatty acid-derived molecules. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13068-021-01965-3. BioMed Central 2021-05-08 /pmc/articles/PMC8106135/ /pubmed/33964988 http://dx.doi.org/10.1186/s13068-021-01965-3 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Zhang, Yang Peng, Jie Zhao, Huimin Shi, Shuobo Engineering oleaginous yeast Rhodotorula toruloides for overproduction of fatty acid ethyl esters |
title | Engineering oleaginous yeast Rhodotorula toruloides for overproduction of fatty acid ethyl esters |
title_full | Engineering oleaginous yeast Rhodotorula toruloides for overproduction of fatty acid ethyl esters |
title_fullStr | Engineering oleaginous yeast Rhodotorula toruloides for overproduction of fatty acid ethyl esters |
title_full_unstemmed | Engineering oleaginous yeast Rhodotorula toruloides for overproduction of fatty acid ethyl esters |
title_short | Engineering oleaginous yeast Rhodotorula toruloides for overproduction of fatty acid ethyl esters |
title_sort | engineering oleaginous yeast rhodotorula toruloides for overproduction of fatty acid ethyl esters |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8106135/ https://www.ncbi.nlm.nih.gov/pubmed/33964988 http://dx.doi.org/10.1186/s13068-021-01965-3 |
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