Engineering of <i>Saccharomyces cerevisiae</i> for 24-Methylene-Cholesterol Production

24-Methylene-cholesterol is a necessary substrate for the biosynthesis of physalin and withanolide, which show promising anticancer activities. It is difficult and costly to prepare 24-methylene-cholesterol via total chemical synthesis. In this study, we engineered the biosynthesis of 24-methylene-c...

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Autores principales:	Jiao Yang, Changfu Li, Yansheng Zhang
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	24-methylene-cholesterol campesterol 7-dehydrocholesterol reductase Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/03e752b95b7d42dfb53b99508cb43ed7
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spelling	oai:doaj.org-article:03e752b95b7d42dfb53b99508cb43ed72021-11-25T16:54:08ZEngineering of <i>Saccharomyces cerevisiae</i> for 24-Methylene-Cholesterol Production10.3390/biom111117102218-273Xhttps://doaj.org/article/03e752b95b7d42dfb53b99508cb43ed72021-11-01T00:00:00Zhttps://www.mdpi.com/2218-273X/11/11/1710https://doaj.org/toc/2218-273X24-Methylene-cholesterol is a necessary substrate for the biosynthesis of physalin and withanolide, which show promising anticancer activities. It is difficult and costly to prepare 24-methylene-cholesterol via total chemical synthesis. In this study, we engineered the biosynthesis of 24-methylene-cholesterol in <i>Saccharomyces cerevisiae</i> by disrupting the two enzymes (i.e., ERG4 and ERG5) in the yeast’s native ergosterol pathway, with ERG5 being replaced with the DHCR7 (7-dehydrocholesterol reductase) enzyme. Three versions of DHCR7 originating from different organisms—including the DHCR7 from <i>Physalis angulata</i> (PhDHCR7) newly discovered in this study, as well as the previously reported OsDHCR7 from <i>Oryza sativa</i> and XlDHCR7 from <i>Xenopus laevis</i>—were assessed for their ability to produce 24-methylene-cholesterol. XlDHCR7 showed the best performance, producing 178 mg/L of 24-methylene-cholesterol via flask-shake cultivation. The yield could be increased up to 225 mg/L, when one additional copy of the <i>XlDHCR7</i> expression cassette was integrated into the yeast genome. The 24-methylene-cholesterol-producing strain obtained in this study could serve as a platform for characterizing the downstream enzymes involved in the biosynthesis of physalin or withanolide, given that 24-methylene-cholesterol is a common precursor of these chemicals.Jiao YangChangfu LiYansheng ZhangMDPI AGarticle24-methylene-cholesterolcampesterol7-dehydrocholesterol reductaseMicrobiologyQR1-502ENBiomolecules, Vol 11, Iss 1710, p 1710 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	24-methylene-cholesterol campesterol 7-dehydrocholesterol reductase Microbiology QR1-502
spellingShingle	24-methylene-cholesterol campesterol 7-dehydrocholesterol reductase Microbiology QR1-502 Jiao Yang Changfu Li Yansheng Zhang Engineering of <i>Saccharomyces cerevisiae</i> for 24-Methylene-Cholesterol Production
description	24-Methylene-cholesterol is a necessary substrate for the biosynthesis of physalin and withanolide, which show promising anticancer activities. It is difficult and costly to prepare 24-methylene-cholesterol via total chemical synthesis. In this study, we engineered the biosynthesis of 24-methylene-cholesterol in <i>Saccharomyces cerevisiae</i> by disrupting the two enzymes (i.e., ERG4 and ERG5) in the yeast’s native ergosterol pathway, with ERG5 being replaced with the DHCR7 (7-dehydrocholesterol reductase) enzyme. Three versions of DHCR7 originating from different organisms—including the DHCR7 from <i>Physalis angulata</i> (PhDHCR7) newly discovered in this study, as well as the previously reported OsDHCR7 from <i>Oryza sativa</i> and XlDHCR7 from <i>Xenopus laevis</i>—were assessed for their ability to produce 24-methylene-cholesterol. XlDHCR7 showed the best performance, producing 178 mg/L of 24-methylene-cholesterol via flask-shake cultivation. The yield could be increased up to 225 mg/L, when one additional copy of the <i>XlDHCR7</i> expression cassette was integrated into the yeast genome. The 24-methylene-cholesterol-producing strain obtained in this study could serve as a platform for characterizing the downstream enzymes involved in the biosynthesis of physalin or withanolide, given that 24-methylene-cholesterol is a common precursor of these chemicals.
format	article
author	Jiao Yang Changfu Li Yansheng Zhang
author_facet	Jiao Yang Changfu Li Yansheng Zhang
author_sort	Jiao Yang
title	Engineering of <i>Saccharomyces cerevisiae</i> for 24-Methylene-Cholesterol Production
title_short	Engineering of <i>Saccharomyces cerevisiae</i> for 24-Methylene-Cholesterol Production
title_full	Engineering of <i>Saccharomyces cerevisiae</i> for 24-Methylene-Cholesterol Production
title_fullStr	Engineering of <i>Saccharomyces cerevisiae</i> for 24-Methylene-Cholesterol Production
title_full_unstemmed	Engineering of <i>Saccharomyces cerevisiae</i> for 24-Methylene-Cholesterol Production
title_sort	engineering of <i>saccharomyces cerevisiae</i> for 24-methylene-cholesterol production
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/03e752b95b7d42dfb53b99508cb43ed7
work_keys_str_mv	AT jiaoyang engineeringofisaccharomycescerevisiaeifor24methylenecholesterolproduction AT changfuli engineeringofisaccharomycescerevisiaeifor24methylenecholesterolproduction AT yanshengzhang engineeringofisaccharomycescerevisiaeifor24methylenecholesterolproduction
_version_	1718412816151478272

Engineering of <i>Saccharomyces cerevisiae</i> for 24-Methylene-Cholesterol Production

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