Production of 21-hydroxy-20-methyl-pregna-1,4-dien-3-one by modifying multiple genes in Mycolicibacterium

doi:10.1007/s00253-023-12399-2

	Production of 21-hydroxy-20-methyl-pregna-1,4-dien-3-one by modifying multiple genes in Mycolicibacterium
	Yuan, Chenyang1,2,3 ; Ma, Zhiguo 1,3; Li, Yixin 4; Zhang, Jingxian 1,3; Liu, Xiangcen 1,3; Han, Suwan 1,3; Du, Guilin1,2,3 ; Shi, Jiping1,2,3 ; Sun, Junsong1,2,3 ; Zhang, Baoguo 1,3
	2023
发表期刊	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
ISSN	0175-7598
EISSN	1432-0614
卷号	107 期号:5-6 页码:1563-1574
发表状态	已发表
DOI	10.1007/s00253-023-12399-2
摘要	Abstract: C22 steroid drug intermediates are suitable for corticosteroids synthesis, and the production of C22 steroids is unsatisfactory due to the intricate steroid metabolism. Among the C22 steroids, 21-hydroxy-20-methyl-pregna-1,4-dien-3-one (1,4-HP) could be used for Δ1-steroid drug synthesis, such as prednisolone. Nevertheless, the production of 1,4-HP remains unsatisfactory. In this study, an ideal 1,4-HP producing strain was constructed. By the knockout of 3-ketosteroid-9-hydroxylase (KshA) genes and 17β-hydroxysteroid dehydrogenase (Hsd4A) gene, the steroid nucleus degradation and the accumulation of C19 steroids in Mycolicibacterium neoaurum were blocked. The mutant strain could transform phytosterols into 1,4-HP as the main product and 21-hydroxy-20-methyl-pregna-4-ene-3-one as a by-product. Subsequently, the purity of 1,4-HP improved to 95.2% by the enhancement of 3-ketosteroid-Δ1-dehydrogenase (KSTD) activity, and the production of 1,4-HP was improved by overexpressing NADH oxidase (NOX) and catalase (KATE) genes. Consequently, the yield of 1,4-HP achieved 10.5 g/L. The molar yield and the purity of 1,4-HP were optimal so far, and the production of 1,4-HP provides a new intermediate for the pharmaceutical steroid industry. Key points: • A third 3-ketosteroid-9-hydroxylase was identified in Mycolicibacterium neoaurum. • An 1,4-HP producer was constructed by KshA and Hsd4A deficiency. • The production of 1,4-HP was improved by KSTD, NOX, and KATE overexpression. © 2023, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
关键词	Alcohols Lipids 21-hydroxy-20-methyl-pregnum-1,4-dien-3-one C22 steroid Drug synthesis Hydroxylases Multiple genes Mycolicibacteria NADH oxidase Phytosterol Prednisolone Steroids drugs
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收录类别	EI ; SCI ; SCOPUS
语种	英语
资助项目	National Key R&D Program of China[2017YFE0112700]
WOS研究方向	Biotechnology & Applied Microbiology
WOS类目	Biotechnology & Applied Microbiology
WOS记录号	WOS:000925681900001
出版者	Springer Science and Business Media Deutschland GmbH
EI入藏号	20230613546881
EI主题词	Genes
EI分类号	461.2 Biological Materials and Tissue Engineering ; 804.1 Organic Compounds
原始文献类型	Article in Press
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/281971
专题	生命科学与技术学院_博士生生命科学与技术学院_特聘教授组_孙俊松组生命科学与技术学院_特聘教授组_史吉平组
通讯作者	Zhang, Baoguo
作者单位	1.Chinese Acad Sci, Shanghai Adv Res Inst, Lab Biorefinery, 99 Haike Rd, Shanghai 201210, Peoples R China 2.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Dept Biol, Waterville, ME 04901 USA
第一作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Yuan, Chenyang,Ma, Zhiguo,Li, Yixin,et al. Production of 21-hydroxy-20-methyl-pregna-1,4-dien-3-one by modifying multiple genes in Mycolicibacterium[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2023,107(5-6):1563-1574.
APA	Yuan, Chenyang.,Ma, Zhiguo.,Li, Yixin.,Zhang, Jingxian.,Liu, Xiangcen.,...&Zhang, Baoguo.(2023).Production of 21-hydroxy-20-methyl-pregna-1,4-dien-3-one by modifying multiple genes in Mycolicibacterium.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,107(5-6),1563-1574.
MLA	Yuan, Chenyang,et al."Production of 21-hydroxy-20-methyl-pregna-1,4-dien-3-one by modifying multiple genes in Mycolicibacterium".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 107.5-6(2023):1563-1574.