Sources and control of impurity during one-pot enzymatic production of dehydroepiandrosterone.
Appl Microbiol Biotechnol
; 108(1): 399, 2024 Jun 29.
Article
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| MEDLINE
| ID: mdl-38951177
ABSTRACT
Dehydroepiandrosterone (DHEA) has a promising market due to its capacity to regulate human hormone levels as well as preventing and treating various diseases. We have established a chemical esterification coupled biocatalytic-based scheme by lipase-catalyzed 4-androstene-3,17-dione (4-AD) hydrolysis to obtain the intermediate product 5-androstene-3,17-dione (5-AD), which was then asymmetrically reduced by a ketoreductase from Sphingomonas wittichii (SwiKR). Co-enzyme required for KR is regenerated by a glucose dehydrogenase (GDH) from Bacillus subtilis. This scheme is more environmentally friendly and more efficient than the current DHEA synthesis pathway. However, a significant amount of 4-AD as by-product was detected during the catalytic process. Focused on the control of by-products, we investigated the source of 4-AD and identified that it is mainly derived from the isomerization activity of SwiKR and GDH. Increasing the proportion of glucose in the catalytic system as well as optimizing the catalytic conditions drastically reduced 4-AD from 24.7 to 6.5% of total substrate amount, and the final yield of DHEA achieved 40.1 g/L. Furthermore, this is the first time that both SwiKR and GDH have been proved to be promiscuous enzymes with dehydrogenase and ketosteroid isomerase (KSI) activities, expanding knowledge of the substrate diversity of the short-chain dehydrogenase family enzymes. KEY POINTS ⢠A strategy of coupling lipase, ketoreductase, and glucose dehydrogenase in producing DHEA from 4-AD ⢠Both SwiKR and GDH are identified with ketosteroid isomerase activity. ⢠Development of catalytic strategy to control by-product and achieve highly selective DHEA production.
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Bases de datos:
MEDLINE
Asunto principal:
Deshidroepiandrosterona
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Sphingomonas
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Lipasa
Idioma:
En
Revista:
Appl Microbiol Biotechnol
Año:
2024
Tipo del documento:
Article
País de afiliación:
China