Highly efficient enzymatic synthesis of tert -butyl ( S )-6-chloro-5-hydroxy-3-oxohexanoate with a mutant alcohol dehydrogenase of Lactobacillus kefir
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| 024 | 7 | 0 | |a 10.1007/s00253-015-6675-1 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6675-1 | ||
| 245 | 0 | 0 | |a Highly efficient enzymatic synthesis of tert -butyl ( S )-6-chloro-5-hydroxy-3-oxohexanoate with a mutant alcohol dehydrogenase of Lactobacillus kefir |h [Elektronische Daten] |c [Xiu-Juan He, Shao-Yun Chen, Jian-Ping Wu, Li-Rong Yang, Gang Xu] |
| 520 | 3 | |a tert-Butyl (S)-6-chloro-5-hydroxy-3-oxohexanoate ((S)-CHOH) is a valuable chiral synthon, which is used for the synthesis of the cholesterol-lowering drugs atorvastatin and rosuvastatin. To date, only the alcohol dehydrogenases from Lactobacillus brevis (LbADH) and Lactobacillus kefir (LkADH) have demonstrated catalytic activity toward the asymmetric reduction of tert-butyl 6-chloro-3,5-dioxohexanoate (CDOH) to (S)-CHOH. Herein, a tetrad mutant of LkADH (LkTADH), A94T/F147L/L199H/A202L, was screened to be more efficient in this bioreduction process, exhibiting a 3.7- and 42-fold improvement in specific activity toward CDOH (1.27U/mg) over LbADH (0.34U/mg) and wild-type LkADH (0.03U/mg), respectively. The molecular basis for the improved catalytic activity of LkTADH toward CDOH was investigated using homology modeling and docking analysis. Two major issues had a significant impact on the biocatalytic efficiency of this process, including (i) the poor aqueous stability of the substrate and (ii) partial substrate inhibition. A fed-batch strategy was successfully developed to address these issues and maintain a suitably low substrate concentration throughout the entire process. Several other parameters were also optimized, including the pH, temperature, NADP+ concentration and cell loading. A final CDOH concentration of 427mM (100g/L) gave (S)-CHOH in 94% yield and 99.5% e.e. after a reaction time of 38h with whole cells expressing LkTADH. The space-time yield and turnover number of NADP+ in this process were 10.6mmol/L/h and 16,060mol/mol, respectively, which were the highest values ever reported. This new approach therefore represents a promising alternative for the efficient synthesis of (S)-CHOH. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a Lactobacillus kefir alcohol dehydrogenase |2 nationallicence | |
| 690 | 7 | |a tert -Butyl ( S )-6-chloro-5-hydroxy-3-oxohexanoate |2 nationallicence | |
| 690 | 7 | |a Statins |2 nationallicence | |
| 690 | 7 | |a Engineered enzyme |2 nationallicence | |
| 690 | 7 | |a Fed-batch strategy |2 nationallicence | |
| 700 | 1 | |a He |D Xiu-Juan |u Institute of Bioengineering, College of Chemical and Biochemical Engineering, Zhejiang University, 38# Zheda Road, 310027, Hangzhou, People's Republic of China |4 aut | |
| 700 | 1 | |a Chen |D Shao-Yun |u College of Life Science, Zhejiang Chinese Medical University, 548# Binwen Road, 310053, Hangzhou, People's Republic of China |4 aut | |
| 700 | 1 | |a Wu |D Jian-Ping |u Institute of Bioengineering, College of Chemical and Biochemical Engineering, Zhejiang University, 38# Zheda Road, 310027, Hangzhou, People's Republic of China |4 aut | |
| 700 | 1 | |a Yang |D Li-Rong |u Institute of Bioengineering, College of Chemical and Biochemical Engineering, Zhejiang University, 38# Zheda Road, 310027, Hangzhou, People's Republic of China |4 aut | |
| 700 | 1 | |a Xu |D Gang |u Institute of Bioengineering, College of Chemical and Biochemical Engineering, Zhejiang University, 38# Zheda Road, 310027, Hangzhou, People's Republic of China |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/21(2015-11-01), 8963-8975 |x 0175-7598 |q 99:21<8963 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6675-1 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-015-6675-1 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a He |D Xiu-Juan |u Institute of Bioengineering, College of Chemical and Biochemical Engineering, Zhejiang University, 38# Zheda Road, 310027, Hangzhou, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Chen |D Shao-Yun |u College of Life Science, Zhejiang Chinese Medical University, 548# Binwen Road, 310053, Hangzhou, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Wu |D Jian-Ping |u Institute of Bioengineering, College of Chemical and Biochemical Engineering, Zhejiang University, 38# Zheda Road, 310027, Hangzhou, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Yang |D Li-Rong |u Institute of Bioengineering, College of Chemical and Biochemical Engineering, Zhejiang University, 38# Zheda Road, 310027, Hangzhou, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Xu |D Gang |u Institute of Bioengineering, College of Chemical and Biochemical Engineering, Zhejiang University, 38# Zheda Road, 310027, Hangzhou, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/21(2015-11-01), 8963-8975 |x 0175-7598 |q 99:21<8963 |1 2015 |2 99 |o 253 | ||