Promoter engineering-mediated Tuning of esterase and transaminase expression for the chemoenzymatic synthesis of sitagliptin phosphate at the kilogram-scale
| Autor: | Jeong Ho Kim, Amol D. Pagar, Hyungdon Yun, Seongseon Yu, Hyunwoo Jeon, Taresh P. Khobragade, Hyunsang Jung, Sharad Sarak, Sung Ryoung Lee, Heung Mo Kang, Geon-Hee Kim, Somin Lee, Pritam Giri, Lee Myeong Seok, Sung‐Su Cho, Mahesh D. Patil, Hye‐Jung Park, Seong‐Hwa Park, In Suk Choi |
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| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
0301 basic medicine Bioengineering 01 natural sciences Applied Microbiology and Biotechnology Esterase Sitagliptin Phosphate Transaminase 03 medical and health sciences Biotransformation 010608 biotechnology medicine Escherichia coli Promoter Regions Genetic Transaminases chemistry.chemical_classification Chromatography Esterases Substrate (chemistry) 030104 developmental biology Enzyme chemistry Sitagliptin Yield (chemistry) Microorganisms Genetically-Modified Genetic Engineering Biotechnology medicine.drug |
| Zdroj: | Biotechnology and bioengineeringREFERENCES. 118(8) |
| ISSN: | 1097-0290 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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