Enzymatic characterization of a recombinant carbonyl reductase from Acetobacter sp. CCTCC M209061
Abstract Background Acetobacter sp. CCTCC M209061 could catalyze carbonyl compounds to chiral alcohols following anti-Prelog rule with excellent enantioselectivity. Therefore, the enzymatic characterization of carbonyl reductase (CR) from Acetobacter sp. CCTCC M209061 needs to be investigated. Resul...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2017-08-01
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| Series: | Bioresources and Bioprocessing |
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| Online Access: | http://link.springer.com/article/10.1186/s40643-017-0169-1 |
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| author | Ping Wei Yu-Han Cui Min-Hua Zong Pei Xu Jian Zhou Wen-Yong Lou |
| author_facet | Ping Wei Yu-Han Cui Min-Hua Zong Pei Xu Jian Zhou Wen-Yong Lou |
| author_sort | Ping Wei |
| collection | DOAJ |
| description | Abstract Background Acetobacter sp. CCTCC M209061 could catalyze carbonyl compounds to chiral alcohols following anti-Prelog rule with excellent enantioselectivity. Therefore, the enzymatic characterization of carbonyl reductase (CR) from Acetobacter sp. CCTCC M209061 needs to be investigated. Results A CR from Acetobacter sp. CCTCC M209061 (AcCR) was cloned and expressed in E. coli. AcCR was purified and characterized, finding that AcCR as a dual coenzyme-dependent short-chain dehydrogenase/reductase (SDR) was more preferred to NADH for biocatalytic reactions. The AcCR was activated and stable when the temperature was under 35 °C and the pH range was from 6.0 to 8.0 for the reduction of 4′-chloroacetophenone with NADH as coenzyme, and the optimal temperature and pH were 45 °C and 8.5, respectively, for the oxidation reaction of isopropanol with NAD+. The enzyme showed moderate thermostability with half-lives of 25.75 h at 35 °C and 13.93 h at 45 °C, respectively. Moreover, the AcCR has broad substrate specificity to a range of ketones and ketoesters, and could catalyze to produce chiral alcohol with e.e. >99% for the majority of tested substrates following the anti-Prelog rule. Conclusions The recombinant AcCR exhibited excellent enantioselectivity, broad substrate spectrum, and highly stereoselective anti-Prelog reduction of prochiral ketones. These results suggest that AcCR is a powerful catalyst for the production of anti-Prelog alcohols. Graphical abstract The biocatalytic reactions conducted with the recombinant AcCR |
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| id | doaj.art-3f0efdc03b1c4d9e98a59eb32bd9895e |
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| language | English |
| last_indexed | 2024-04-13T06:58:46Z |
| publishDate | 2017-08-01 |
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| series | Bioresources and Bioprocessing |
| spelling | doaj.art-3f0efdc03b1c4d9e98a59eb32bd9895e2022-12-22T02:57:11ZengSpringerOpenBioresources and Bioprocessing2197-43652017-08-014111210.1186/s40643-017-0169-1Enzymatic characterization of a recombinant carbonyl reductase from Acetobacter sp. CCTCC M209061Ping Wei0Yu-Han Cui1Min-Hua Zong2Pei Xu3Jian Zhou4Wen-Yong Lou5Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of TechnologyLab of Applied Biocatalysis, School of Food Science and Engineering, South China University of TechnologyLab of Applied Biocatalysis, School of Food Science and Engineering, South China University of TechnologyLab of Applied Biocatalysis, School of Food Science and Engineering, South China University of TechnologySchool of Chemistry and Chemical Engineering, South China University of TechnologyLab of Applied Biocatalysis, School of Food Science and Engineering, South China University of TechnologyAbstract Background Acetobacter sp. CCTCC M209061 could catalyze carbonyl compounds to chiral alcohols following anti-Prelog rule with excellent enantioselectivity. Therefore, the enzymatic characterization of carbonyl reductase (CR) from Acetobacter sp. CCTCC M209061 needs to be investigated. Results A CR from Acetobacter sp. CCTCC M209061 (AcCR) was cloned and expressed in E. coli. AcCR was purified and characterized, finding that AcCR as a dual coenzyme-dependent short-chain dehydrogenase/reductase (SDR) was more preferred to NADH for biocatalytic reactions. The AcCR was activated and stable when the temperature was under 35 °C and the pH range was from 6.0 to 8.0 for the reduction of 4′-chloroacetophenone with NADH as coenzyme, and the optimal temperature and pH were 45 °C and 8.5, respectively, for the oxidation reaction of isopropanol with NAD+. The enzyme showed moderate thermostability with half-lives of 25.75 h at 35 °C and 13.93 h at 45 °C, respectively. Moreover, the AcCR has broad substrate specificity to a range of ketones and ketoesters, and could catalyze to produce chiral alcohol with e.e. >99% for the majority of tested substrates following the anti-Prelog rule. Conclusions The recombinant AcCR exhibited excellent enantioselectivity, broad substrate spectrum, and highly stereoselective anti-Prelog reduction of prochiral ketones. These results suggest that AcCR is a powerful catalyst for the production of anti-Prelog alcohols. Graphical abstract The biocatalytic reactions conducted with the recombinant AcCRhttp://link.springer.com/article/10.1186/s40643-017-0169-1Carbonyl reductaseAcetobacter sp.Chiral alcoholsEnzymatic characterization |
| spellingShingle | Ping Wei Yu-Han Cui Min-Hua Zong Pei Xu Jian Zhou Wen-Yong Lou Enzymatic characterization of a recombinant carbonyl reductase from Acetobacter sp. CCTCC M209061 Bioresources and Bioprocessing Carbonyl reductase Acetobacter sp. Chiral alcohols Enzymatic characterization |
| title | Enzymatic characterization of a recombinant carbonyl reductase from Acetobacter sp. CCTCC M209061 |
| title_full | Enzymatic characterization of a recombinant carbonyl reductase from Acetobacter sp. CCTCC M209061 |
| title_fullStr | Enzymatic characterization of a recombinant carbonyl reductase from Acetobacter sp. CCTCC M209061 |
| title_full_unstemmed | Enzymatic characterization of a recombinant carbonyl reductase from Acetobacter sp. CCTCC M209061 |
| title_short | Enzymatic characterization of a recombinant carbonyl reductase from Acetobacter sp. CCTCC M209061 |
| title_sort | enzymatic characterization of a recombinant carbonyl reductase from acetobacter sp cctcc m209061 |
| topic | Carbonyl reductase Acetobacter sp. Chiral alcohols Enzymatic characterization |
| url | http://link.springer.com/article/10.1186/s40643-017-0169-1 |
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