Characterization of a robust glucose 1-dehydrogenase, SyGDH, and its application in NADPH regeneration for the asymmetric reduction of haloketone by a carbonyl reductase in organic solvent/buffer system

2020 ◽  
Vol 89 ◽  
pp. 55-62
Author(s):  
Die Hu ◽  
Zheng Wen ◽  
Chuang Li ◽  
Bochun Hu ◽  
Ting Zhang ◽  
...  
Keyword(s):  
Organic Solvent ◽  
Asymmetric Reduction ◽  
Carbonyl Reductase ◽  
Buffer System ◽  
Nadph Regeneration

scholarly journals Characterization of a Carbonyl Reductase from Rhodococcus erythropolis WZ010 and Its Variant Y54F for Asymmetric Synthesis of (S)-N-Boc-3-Hydroxypiperidine

Molecules ◽  
2018 ◽  
Vol 23 (12) ◽  
pp. 3117 ◽  
Author(s):  
Xiangxian Ying ◽  
Jie Zhang ◽  
Can Wang ◽  
Meijuan Huang ◽  
Yuting Ji ◽  
...  
Keyword(s):  
Asymmetric Reduction ◽  
Rhodococcus Erythropolis ◽  
Catalytic Efficiency ◽  
Carbonyl Reductase ◽  
Specific Enzyme ◽  
Practical Applications ◽  
Whole Cells ◽  
E Coli ◽  
Irreversible Reduction

The recombinant carbonyl reductase from Rhodococcus erythropolis WZ010 (ReCR) demonstrated strict (S)-stereoselectivity and catalyzed the irreversible reduction of N-Boc-3-piperidone (NBPO) to (S)-N-Boc-3-hydroxypiperidine [(S)-NBHP], a key chiral intermediate in the synthesis of ibrutinib. The NAD(H)-specific enzyme was active within broad ranges of pH and temperature and had remarkable activity in the presence of higher concentration of organic solvents. The amino acid residue at position 54 was critical for the activity and the substitution of Tyr54 to Phe significantly enhanced the catalytic efficiency of ReCR. The kcat/Km values of ReCR Y54F for NBPO, (R/S)-2-octanol, and 2-propanol were 49.17 s−1 mM−1, 56.56 s−1 mM−1, and 20.69 s−1 mM−1, respectively. In addition, the (S)-NBHP yield was as high as 95.92% when whole cells of E. coli overexpressing ReCR variant Y54F catalyzed the asymmetric reduction of 1.5 M NBPO for 12 h in the aqueous/(R/S)-2-octanol biphasic system, demonstrating the great potential of ReCR variant Y54F for practical applications.

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