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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...

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Autores principales: Du, Hao-Xing, Xiao, Wen-Hai, Wang, Ying, Zhou, Xiao, Zhang, Yu, Liu, Duo, Yuan, Ying-Jin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
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.
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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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