Promoter engineering-mediated Tuning of esterase and transaminase expression for the chemoenzymatic synthesis of sitagliptin phosphate at the kilogram-scale.
Biotechnol Bioeng
; 118(8): 3263-3268, 2021 08.
Article
in En
| MEDLINE
| ID: mdl-33990942
ABSTRACT
Here, we report a bienzymatic cascade to produce ß-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 ß-amino acid (229 mM). Sitagliptin phosphate was chemically synthesized from ß-amino acids with 82% yield and > 99% purity.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
Genetic Engineering
/
Promoter Regions, Genetic
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Escherichia coli
/
Esterases
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Sitagliptin Phosphate
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Microorganisms, Genetically-Modified
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Transaminases
Language:
En
Journal:
Biotechnol Bioeng
Year:
2021
Type:
Article
Affiliation country:
South Korea