http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
- 中文 을 입력하시려면 zhongwen을 입력하시고 space를누르시면됩니다.
- 北京 을 입력하시려면 beijing을 입력하시고 space를 누르시면 됩니다.
RISS 인기검색어
- 검색키워드
- 저자 : SARAKSHARADCHANGDEO (검색결과 2 건)
- 무료
- 기관 내 무료
- 유료
-
이소민,전현우,Pritam Giri,이욱재,정현상,임성아,SARAKSHARADCHANGDEO,Taresh P. Khobragade,김병기,윤형돈 한국생물공학회 2021 Biotechnology and Bioprocess Engineering Vol.26 No.3
Amine dehydrogenases (AmDHs) are one of the key emerging enzymes used in the synthesis of various amines with the expense of only one ammonium ion as an amino donor, thereby, generates only water molecule as a by-product. Currently, most available AmDHs have been created through protein engineering using the existing natural L-amino acid dehydrogenase, and native AmDHs are rarely reported. In this study, a novel native AmDH from Laribacter hongkongensis (LhAmDH) was identified based on the GenBank database using a sequence-driven approach. LhAmDH showed a good activity towards various carbonyl compounds such as cyclohexanone (170 mU/mg) and isovaleraldehyde (214 mU/mg). The reductive amination of model substrate, cyclohexanone (up to 100 mM) into cyclohexylamine was successfully performed in LhAmDH and FDH system with > 99% conversion using Escherichia coli whole-cell as well as purified enzymes. Furthermore, three enzymes cascade (ω-transaminase, LhAmDH, and FDH) was designed to produce chiral amine from the corresponding ketone using inexpensive ammonium formate as sole sacrificial agent. The active site of LhAmDH residues were predicted using the protein structure homology model building program SWISS-MODEL server. In the docking analysis, cyclohexanone is well-orientated with -5.4 kcal/mol of binding energy and 3.16 Å distance from side chain of E104, which is a key residue for interacting ammonia and substrate. This LhAmDH model can be used as a promising template to produce chiral amines through semi-rational design.
-
Biocatalytic Cascade for Synthesis of Sitagliptin Intermediate Employing Coupled Transaminase
Taresh P. Khobragade,Pagar Amol D.,GIRI PRITAM DEVIDAS,SARAKSHARADCHANGDEO,전현우,주상우,고영환,박부수,윤형돈 한국생물공학회 2023 Biotechnology and Bioprocess Engineering Vol.28 No.2
Transaminases (TAs) are employed in synthesizing various enantiopure β‐amino acids, key precursors for various pharmaceuticals. Sitagliptin, an oral hyperglycemic drug, is a well-known example. Herein, we developed the coupled enzyme cascade to synthesize the sitagliptin intermediate by fusing two different TAs to regenerate the amino donor. In a cascade system, ethyl 3-oxo-4-(2,4,5- trifluorophenyl) butanoate (1) was converted by esterase from Pseudomonas stutzeri (EstPS) to respective β-keto acid (2) which was subsequently converted by first TA to sitagliptin intermediate (3) using (S)-α-MBA as an amino donor and the acetophenone formed in the reaction was recycled by the second TA to (S)-α-MBA. A single wholecell system was established by the co-expression of esterase and TA fusion protein. The whole-cell biotransformation reaction was performed with varying substrate concentrations from 50-200 mM. The excellent conversion of the product was achieved, ranging from 62-to 100% at the expense of only 25 mM (S)-α-MBA. Notably, our designed system with fusion protein can produce ~5-fold higher product at the expense of 0.5 equivalent (S)-α-MBA. Finally, a preparative scale reaction was performed with 98% conversion.
연관 검색어 추천
이 검색어로 많이 본 자료
활용도 높은 자료
