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03236nam a2200421Ia 4500 |
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10-1186-s12934-022-01786-5 |
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|a 14752859 (ISSN)
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|a One-pot biosynthesis of 7β-hydroxyandrost-4-ene-3,17-dione from phytosterols by cofactor regeneration system in engineered mycolicibacterium neoaurum
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|b BioMed Central Ltd
|c 2022
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|z View Fulltext in Publisher
|u https://doi.org/10.1186/s12934-022-01786-5
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|a Background: 7β-hydroxylated steroids (7β-OHSt) possess significant activities in anti-inflammatory and neuroprotection, and some of them have been widely used in clinics. However, the production of 7β-OHSt is still a challenge due to the lack of cheap 7β-hydroxy precursor and the difficulty in regio- and stereo-selectively hydroxylation at the inert C7 site of steroids in industry. The conversion of phytosterols by Mycolicibacterium species to the commercial precursor, androst-4-ene-3,17-dione (AD), is one of the basic ways to produce different steroids. This study presents a way to produce a basic 7β-hydroxy precursor, 7β-hydroxyandrost-4-ene-3,17-dione (7β-OH-AD) in Mycolicibacterium, for 7β-OHSt synthesis. Results: A mutant of P450-BM3, mP450-BM3, was mutated and engineered into an AD producing strain for the efficient production of 7β-OH-AD. The enzyme activity of mP450-BM3 was then increased by 1.38 times through protein engineering and the yield of 7β-OH-AD was increased from 34.24 mg L− 1 to 66.25 mg L− 1. To further enhance the performance of 7β-OH-AD producing strain, the regeneration of nicotinamide adenine dinucleotide phosphate (NADPH) for the activity of mP450-BM3-0 was optimized by introducing an NAD kinase (NADK) and a glucose-6-phosphate dehydrogenase (G6PDH). Finally, the engineered strain could produce 164.52 mg L− 1 7β-OH-AD in the cofactor recycling and regeneration system. Conclusions: This was the first report on the one-pot biosynthesis of 7β-OH-AD from the conversion of cheap phytosterols by an engineered microorganism, and the yield was significantly increased through the mutation of mP450-BM3 combined with overexpression of NADK and G6PDH. The present strategy may be developed as a basic industrial pathway for the commercial production of high value products from cheap raw materials. © 2022, The Author(s).
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|a 7β-hydroxyandrost-4-ene-3,17-dione
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|a biotransformation
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|a Biotransformation
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|a metabolism
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|a mP450-BM3
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|a Mycobacteriaceae
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|a Mycobacteriaceae
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|a Mycolicibacterium
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|a Nicotinamide adenine dinucleotide phosphate regeneration
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|a phytosterol
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|a Phytosterols
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|a Phytosterols
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|a regeneration
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|a Regeneration
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|a steroid
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|a Steroids
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|a Gao, B.
|e author
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|a Ji, W.-T.
|e author
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|a Liu, K.
|e author
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|a Liu, Y.-J.
|e author
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|a Tao, X.-Y.
|e author
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|a Wang, F.-Q.
|e author
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|a Wei, D.-Z.
|e author
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|a Zhao, M.
|e author
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|a Zhao, Y.-Q.
|e author
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|t Microbial Cell Factories
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