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Characterization of an allylic/benzyl alcohol dehydrogenase from Yokenella sp. strain WZY002, an organism potentially useful for the synthesis of α,β-unsaturated alcohols from allylic aldehydes and ketones.
Appl Environ Microbiol. 2014 Apr; 80(8):2399-409.AE

Abstract

A novel whole-cell biocatalyst with high allylic alcohol-oxidizing activities was screened and identified as Yokenella sp. WZY002, which chemoselectively reduced the C=O bond of allylic aldehydes/ketones to the corresponding α,β-unsaturated alcohols at 30°C and pH 8.0. The strain also had the capacity of stereoselectively reducing aromatic ketones to (S)-enantioselective alcohols. The enzyme responsible for the predominant allylic/benzyl alcohol dehydrogenase activity was purified to homogeneity and designated YsADH (alcohol dehydrogenase from Yokenella sp.), which had a calculated subunit molecular mass of 36,411 Da. The gene encoding YsADH was subsequently expressed in Escherichia coli, and the purified recombinant YsADH protein was characterized. The enzyme strictly required NADP(H) as a coenzyme and was putatively zinc dependent. The optimal pH and temperature for crotonaldehyde reduction were pH 6.5 and 65°C, whereas those for crotyl alcohol oxidation were pH 8.0 and 55°C. The enzyme showed moderate thermostability, with a half-life of 6.2 h at 55°C. It was robust in the presence of organic solvents and retained 87.5% of the initial activity after 24 h of incubation with 20% (vol/vol) dimethyl sulfoxide. The enzyme preferentially catalyzed allylic/benzyl aldehydes as the substrate in the reduction of aldehydes/ketones and yielded the highest activity of 427 U mg(-1) for benzaldehyde reduction, while the alcohol oxidation reaction demonstrated the maximum activity of 79.9 U mg(-1) using crotyl alcohol as the substrate. Moreover, kinetic parameters of the enzyme showed lower Km values and higher catalytic efficiency for crotonaldehyde/benzaldehyde and NADPH than for crotyl alcohol/benzyl alcohol and NADP(+), suggesting the nature of being an aldehyde reductase.

Authors+Show Affiliations

College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24509923

Citation

Ying, Xiangxian, et al. "Characterization of an Allylic/benzyl Alcohol Dehydrogenase From Yokenella Sp. Strain WZY002, an Organism Potentially Useful for the Synthesis of Α,β-unsaturated Alcohols From Allylic Aldehydes and Ketones." Applied and Environmental Microbiology, vol. 80, no. 8, 2014, pp. 2399-409.
Ying X, Wang Y, Xiong B, et al. Characterization of an allylic/benzyl alcohol dehydrogenase from Yokenella sp. strain WZY002, an organism potentially useful for the synthesis of α,β-unsaturated alcohols from allylic aldehydes and ketones. Appl Environ Microbiol. 2014;80(8):2399-409.
Ying, X., Wang, Y., Xiong, B., Wu, T., Xie, L., Yu, M., & Wang, Z. (2014). Characterization of an allylic/benzyl alcohol dehydrogenase from Yokenella sp. strain WZY002, an organism potentially useful for the synthesis of α,β-unsaturated alcohols from allylic aldehydes and ketones. Applied and Environmental Microbiology, 80(8), 2399-409. https://doi.org/10.1128/AEM.03980-13
Ying X, et al. Characterization of an Allylic/benzyl Alcohol Dehydrogenase From Yokenella Sp. Strain WZY002, an Organism Potentially Useful for the Synthesis of Α,β-unsaturated Alcohols From Allylic Aldehydes and Ketones. Appl Environ Microbiol. 2014;80(8):2399-409. PubMed PMID: 24509923.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of an allylic/benzyl alcohol dehydrogenase from Yokenella sp. strain WZY002, an organism potentially useful for the synthesis of α,β-unsaturated alcohols from allylic aldehydes and ketones. AU - Ying,Xiangxian, AU - Wang,Yifang, AU - Xiong,Bin, AU - Wu,Tingting, AU - Xie,Liping, AU - Yu,Meilan, AU - Wang,Zhao, Y1 - 2014/02/07/ PY - 2014/2/11/entrez PY - 2014/2/11/pubmed PY - 2014/12/15/medline SP - 2399 EP - 409 JF - Applied and environmental microbiology JO - Appl. Environ. Microbiol. VL - 80 IS - 8 N2 - A novel whole-cell biocatalyst with high allylic alcohol-oxidizing activities was screened and identified as Yokenella sp. WZY002, which chemoselectively reduced the C=O bond of allylic aldehydes/ketones to the corresponding α,β-unsaturated alcohols at 30°C and pH 8.0. The strain also had the capacity of stereoselectively reducing aromatic ketones to (S)-enantioselective alcohols. The enzyme responsible for the predominant allylic/benzyl alcohol dehydrogenase activity was purified to homogeneity and designated YsADH (alcohol dehydrogenase from Yokenella sp.), which had a calculated subunit molecular mass of 36,411 Da. The gene encoding YsADH was subsequently expressed in Escherichia coli, and the purified recombinant YsADH protein was characterized. The enzyme strictly required NADP(H) as a coenzyme and was putatively zinc dependent. The optimal pH and temperature for crotonaldehyde reduction were pH 6.5 and 65°C, whereas those for crotyl alcohol oxidation were pH 8.0 and 55°C. The enzyme showed moderate thermostability, with a half-life of 6.2 h at 55°C. It was robust in the presence of organic solvents and retained 87.5% of the initial activity after 24 h of incubation with 20% (vol/vol) dimethyl sulfoxide. The enzyme preferentially catalyzed allylic/benzyl aldehydes as the substrate in the reduction of aldehydes/ketones and yielded the highest activity of 427 U mg(-1) for benzaldehyde reduction, while the alcohol oxidation reaction demonstrated the maximum activity of 79.9 U mg(-1) using crotyl alcohol as the substrate. Moreover, kinetic parameters of the enzyme showed lower Km values and higher catalytic efficiency for crotonaldehyde/benzaldehyde and NADPH than for crotyl alcohol/benzyl alcohol and NADP(+), suggesting the nature of being an aldehyde reductase. SN - 1098-5336 UR - https://www.unboundmedicine.com/medline/citation/24509923/Characterization_of_an_allylic/benzyl_alcohol_dehydrogenase_from_Yokenella_sp__strain_WZY002_an_organism_potentially_useful_for_the_synthesis_of_αβ_unsaturated_alcohols_from_allylic_aldehydes_and_ketones_ L2 - http://aem.asm.org/cgi/pmidlookup?view=long&pmid=24509923 DB - PRIME DP - Unbound Medicine ER -