Development of whole cell biocatalytic system for asymmetric synthesis of esomeprazole with enhancing coenzyme biosynthesis pathway.
Enzyme Microb Technol
; 179: 110469, 2024 Sep.
Article
in En
| MEDLINE
| ID: mdl-38878426
ABSTRACT
Esomeprazole is the most popular proton pump inhibitor for treating gastroesophageal reflux disease. Previously, a phenylacetone monooxygenase mutant LnPAMOmu15 (LM15) was obtained by protein engineering for asymmetric synthesis of esomeprazole using pyrmetazole as substrate. To scale up the whole cell asymmetric synthesis of esomeprazole and reduce the cost, in this work, an Escherichia coli whole-cell catalyst harboring LM15 and formate dehydrogenase from Burkholderia stabilis 15516 (BstFDH) were constructed through optimized gene assembly patterns. CRISPR/Cas9 mediated insertion of Ptrc promoter in genome was done to enhance the expression of key genes to increase the cellular NADP supply in the whole cell catalyst, by which the amount of externally added NADP+ for the asymmetric synthesis of esomeprazole decreased to 0.05â¯mM from 0.3â¯mM for reducing the cost. After the optimization of reaction conditions in the reactor, the scalable synthesis of esomeprazole was performed using the efficient LM15-BstFDH whole-cell as catalyst, which showed the highest reported space-time yield of 3.28â¯g/L/h with 50â¯mM of pyrmetazole loading. Isolation procedure was conducted to obtain esomeprazole sodium of 99.55â¯% purity and >â¯99.9â¯% ee with 90.1â¯% isolation yield. This work provides the basis for production of enantio-pure esomeprazole via cost-effective whole cell biocatalysis.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Burkholderia
/
Escherichia coli
/
Biocatalysis
/
Esomeprazole
Language:
En
Journal:
Enzyme Microb Technol
Year:
2024
Document type:
Article
Publication country:
EEUU
/
ESTADOS UNIDOS
/
ESTADOS UNIDOS DA AMERICA
/
EUA
/
UNITED STATES
/
UNITED STATES OF AMERICA
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US
/
USA