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Engineering Yarrowia lipolytica for Campesterol Overproduction
Campesterol is an important precursor for many sterol drugs, e.g. progesterone and hydrocortisone. In order to produce campesterol in Yarrowia lipolytica, C-22 desaturase encoding gene ERG5 was disrupted and the heterologous 7-dehydrocholesterol reductase (DHCR7) encoding gene was constitutively exp...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4709189/ https://www.ncbi.nlm.nih.gov/pubmed/26751680 http://dx.doi.org/10.1371/journal.pone.0146773 |
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author | Du, Hao-Xing Xiao, Wen-Hai Wang, Ying Zhou, Xiao Zhang, Yu Liu, Duo Yuan, Ying-Jin |
author_facet | Du, Hao-Xing Xiao, Wen-Hai Wang, Ying Zhou, Xiao Zhang, Yu Liu, Duo Yuan, Ying-Jin |
author_sort | Du, Hao-Xing |
collection | PubMed |
description | Campesterol is an important precursor for many sterol drugs, e.g. progesterone and hydrocortisone. In order to produce campesterol in Yarrowia lipolytica, C-22 desaturase encoding gene ERG5 was disrupted and the heterologous 7-dehydrocholesterol reductase (DHCR7) encoding gene was constitutively expressed. The codon-optimized DHCR7 from Rallus norvegicus, Oryza saliva and Xenapus laevis were explored and the strain with the gene DHCR7 from X. laevis achieved the highest titer of campesterol due to D409 in substrate binding sites. In presence of glucose as the carbon source, higher biomass conversion yield and product yield were achieved in shake flask compared to that using glycerol and sunflower seed oil. Nevertheless, better cell growth rate was observed in medium with sunflower seed oil as the sole carbon source. Through high cell density fed-batch fermentation under carbon source restriction strategy, a titer of 453±24.7 mg/L campesterol was achieved with sunflower seed oil as the carbon source, which is the highest reported microbial titer known. Our study has greatly enhanced campesterol accumulation in Y. lipolytica, providing new insight into producing complex and desired molecules in microbes. |
format | Online Article Text |
id | pubmed-4709189 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-47091892016-01-15 Engineering Yarrowia lipolytica for Campesterol Overproduction Du, Hao-Xing Xiao, Wen-Hai Wang, Ying Zhou, Xiao Zhang, Yu Liu, Duo Yuan, Ying-Jin PLoS One Research Article Campesterol is an important precursor for many sterol drugs, e.g. progesterone and hydrocortisone. In order to produce campesterol in Yarrowia lipolytica, C-22 desaturase encoding gene ERG5 was disrupted and the heterologous 7-dehydrocholesterol reductase (DHCR7) encoding gene was constitutively expressed. The codon-optimized DHCR7 from Rallus norvegicus, Oryza saliva and Xenapus laevis were explored and the strain with the gene DHCR7 from X. laevis achieved the highest titer of campesterol due to D409 in substrate binding sites. In presence of glucose as the carbon source, higher biomass conversion yield and product yield were achieved in shake flask compared to that using glycerol and sunflower seed oil. Nevertheless, better cell growth rate was observed in medium with sunflower seed oil as the sole carbon source. Through high cell density fed-batch fermentation under carbon source restriction strategy, a titer of 453±24.7 mg/L campesterol was achieved with sunflower seed oil as the carbon source, which is the highest reported microbial titer known. Our study has greatly enhanced campesterol accumulation in Y. lipolytica, providing new insight into producing complex and desired molecules in microbes. Public Library of Science 2016-01-11 /pmc/articles/PMC4709189/ /pubmed/26751680 http://dx.doi.org/10.1371/journal.pone.0146773 Text en © 2016 Du et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Du, Hao-Xing Xiao, Wen-Hai Wang, Ying Zhou, Xiao Zhang, Yu Liu, Duo Yuan, Ying-Jin Engineering Yarrowia lipolytica for Campesterol Overproduction |
title | Engineering Yarrowia lipolytica for Campesterol Overproduction |
title_full | Engineering Yarrowia lipolytica for Campesterol Overproduction |
title_fullStr | Engineering Yarrowia lipolytica for Campesterol Overproduction |
title_full_unstemmed | Engineering Yarrowia lipolytica for Campesterol Overproduction |
title_short | Engineering Yarrowia lipolytica for Campesterol Overproduction |
title_sort | engineering yarrowia lipolytica for campesterol overproduction |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4709189/ https://www.ncbi.nlm.nih.gov/pubmed/26751680 http://dx.doi.org/10.1371/journal.pone.0146773 |
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