Promoter engineering-mediated Tuning of esterase and transaminase expression for the chemoenzymatic synthesis of sitagliptin phosphate at the kilogram-scale.
esterase/lipase
promoter engineering
sitagliptin
transaminase
Journal
Biotechnology and bioengineering
ISSN: 1097-0290
Titre abrégé: Biotechnol Bioeng
Pays: United States
ID NLM: 7502021
Informations de publication
Date de publication:
08 2021
08 2021
Historique:
revised:
06
05
2021
received:
22
03
2021
accepted:
09
05
2021
pubmed:
16
5
2021
medline:
18
1
2022
entrez:
15
5
2021
Statut:
ppublish
Résumé
Here, we report a bienzymatic cascade to produce β-amino acids as an intermediate for the synthesis of the leading oral antidiabetic drug, sitagliptin. A whole-cell biotransformation using recombinant Escherichia coli coexpressing a esterase and transaminase were developed, wherein the desired expression level of each enzyme was achieved by promotor engineering. The small-scale reactions (30 ml) performed under optimized conditions at varying amounts of substrate (100-300 mM) resulted in excellent conversions of 82%-95% for the desired product. Finally, a kilogram-scale enzymatic reaction (250 mM substrate, 220 L) was carried out to produce β-amino acid (229 mM). Sitagliptin phosphate was chemically synthesized from β-amino acids with 82% yield and > 99% purity.
Substances chimiques
Transaminases
EC 2.6.1.-
Esterases
EC 3.1.-
Sitagliptin Phosphate
TS63EW8X6F
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3263-3268Informations de copyright
© 2021 Wiley Periodicals LLC.
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