{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,18]],"date-time":"2026-01-18T12:25:00Z","timestamp":1768739100104,"version":"3.49.0"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2021,12,2]],"date-time":"2021-12-02T00:00:00Z","timestamp":1638403200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"Natural Science Foundation of China","doi-asserted-by":"crossref","award":["21808073"],"award-info":[{"award-number":["21808073"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Natural Science Foundation of Fujian Province, China","award":["2021J05058"],"award-info":[{"award-number":["2021J05058"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>The biosynthesis of chiral alcohols has important value and high attention. Aldo\u2013keto reductases (AKRs) mediated reduction of prochiral carbonyl compounds is an interesting way of synthesizing single enantiomers of chiral alcohols due to the high enantio-, chemo- and regioselectivity of the enzymes. However, relatively little research has been done on characterization and apply of AKRs to asymmetric synthesis of chiral alcohols. In this study, the AKR from <jats:italic>Candida tropicalis<\/jats:italic> MYA-3404 (<jats:italic>C<\/jats:italic>. <jats:italic>tropicalis<\/jats:italic> MYA-3404), was mined and characterized. The AKR shown wider optimum temperature and pH. The AKR exhibited varying degrees of catalytic activity for different substrates, suggesting that the AKR can catalyze a variety of substrates. It is worth mentioning that the AKR could catalytic reduction of keto compounds with benzene rings, such as cetophenone and phenoxyacetone. The AKR exhibited activity on <jats:italic>N<\/jats:italic>,<jats:italic>N<\/jats:italic>-dimethyl-3-keto-3-(2-thienyl)-1-propanamine (DKTP), a key intermediate for biosynthesis of the antidepressant drug duloxetine. Besides, the AKR still has high activity whether in a reaction system containing 10%-30% V\/V organic solvent. What\u2019s more, the AKR showed the strongest stability in six common organic solvents, DMSO, acetonitrile, ethyl acetate, isopropanol, ethanol, and methanol. And, it retains more that 70% enzyme activity after 6 hours, suggesting that the AKR has strong solvent tolerance. Furthermore, the protein sequences of the AKR and its homology were compared, and a 3D model of the AKR docking with coenzyme NADPH were constructed. And the important catalytic and binding sites were identified to explore the binding mechanism of the enzyme and its coenzyme. These properties, predominant organic solvents resistance and extensive substrate spectrum, of the AKR making it has potential applications in the pharmaceutical field.<\/jats:p>","DOI":"10.1371\/journal.pone.0260787","type":"journal-article","created":{"date-parts":[[2021,12,2]],"date-time":"2021-12-02T19:41:30Z","timestamp":1638474090000},"page":"e0260787","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["Gene mining, codon optimization and analysis of binding mechanism of an aldo-keto reductase with high activity, better substrate specificity and excellent solvent tolerance"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8194-6003","authenticated-orcid":true,"given":"Wei","family":"Jiang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoli","family":"Fu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weiliang","family":"Wu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2021,12,2]]},"reference":[{"key":"pone.0260787.ref001","first-page":"1","article-title":"Catalytic asymmetric reductive hydroalkylation of enamides and enecarbamates to chiral aliphatic amines","volume":"12","author":"J-W Wang","year":"2021","journal-title":"Nature communications"},{"key":"pone.0260787.ref002","doi-asserted-by":"crossref","first-page":"1665","DOI":"10.1002\/ange.202012445","article-title":"Chiral Bicyclic Imidazole\u2010Catalyzed Acylative Dynamic Kinetic Resolution for the Synthesis of Chiral Phthalidyl Esters","volume":"133","author":"M Zhou","year":"2021","journal-title":"Angewandte Chemie"},{"key":"pone.0260787.ref003","doi-asserted-by":"crossref","first-page":"4578","DOI":"10.1021\/acs.chemrev.0c00002","article-title":"Modern stereoselective synthesis of chiral sulfinyl compounds","volume":"120","author":"Eb Wojaczy\u0144ska","year":"2020","journal-title":"Chemical Reviews"},{"key":"pone.0260787.ref004","doi-asserted-by":"crossref","first-page":"8569","DOI":"10.1002\/ange.202016286","article-title":"Highly enantioselective CuAAc of functional tertiary alcohols featuring an ethynyl group and their kinetic resolution","volume":"133","author":"K Liao","year":"2021","journal-title":"Angewandte Chemie"},{"key":"pone.0260787.ref005","first-page":"1","article-title":"Asymmetric reduction of prochiral aromatic and hetero aromatic ketones using whole-cell of Lactobacillus senmaizukei biocatalyst","author":"NS \u00c7olak","year":"2021","journal-title":"Synthetic Communications"},{"key":"pone.0260787.ref006","doi-asserted-by":"crossref","first-page":"2838","DOI":"10.1021\/jacs.0c12242","article-title":"Diastereo-and Enantioselective Ruthenium-Catalyzed CC Coupling of 1-Arylpropynes and Alcohols: Alkynes as Chiral Allylmetal Precursors in Carbonyl anti-(\u03b1-Aryl) allylation","volume":"143","author":"M Xiang","year":"2021","journal-title":"Journal of the American Chemical Society"},{"key":"pone.0260787.ref007","doi-asserted-by":"crossref","first-page":"1727","DOI":"10.1002\/anie.201812894","article-title":"Stereoinversion of unactivated alcohols by tethered sulfonamides","volume":"58","author":"PT Marcyk","year":"2019","journal-title":"Angewandte Chemie International Edition"},{"key":"pone.0260787.ref008","doi-asserted-by":"crossref","first-page":"4452","DOI":"10.1002\/chem.201806068","article-title":"Understanding the Reactivity and Selectivity of Fluxional Chiral 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pain","volume":"122","author":"C-F Wang","year":"2016","journal-title":"Anesthesia & Analgesia"},{"key":"pone.0260787.ref012","doi-asserted-by":"crossref","first-page":"4411","DOI":"10.1021\/acs.orglett.9b00950","article-title":"Enantioselective reduction of \u03b1, \u03b2-unsaturated ketones and aryl ketones by perakine reductase","volume":"21","author":"S Cai","year":"2019","journal-title":"Organic letters"},{"key":"pone.0260787.ref013","doi-asserted-by":"crossref","first-page":"2483","DOI":"10.1021\/la950348l","article-title":"Adsorption of chiral alcohols on \u201cchiral\u201d metal surfaces","volume":"12","author":"CF McFadden","year":"1996","journal-title":"Langmuir"},{"key":"pone.0260787.ref014","doi-asserted-by":"crossref","first-page":"11505","DOI":"10.1002\/ange.202003104","article-title":"Asymmetric Guerbet Reaction to Access Chiral Alcohols","volume":"132","author":"K Wang","year":"2020","journal-title":"Angewandte 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stereoselectivity","volume":"89","author":"Y Ni","year":"2011","journal-title":"Applied microbiology and biotechnology"},{"key":"pone.0260787.ref018","doi-asserted-by":"crossref","first-page":"e0137744","DOI":"10.1371\/journal.pone.0137744","article-title":"A novel aldo-keto reductase (AKR17A1) of Anabaena sp. 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Saccharomyces cerevisiae reductase with high enantioselectivity","volume":"87","author":"YH Choi","year":"2010","journal-title":"Applied microbiology and biotechnology"},{"key":"pone.0260787.ref022","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1128\/AEM.71.2.1101-1104.2005","article-title":"Purification and cDNA cloning of NADPH-dependent aldoketoreductase, involved in asymmetric reduction of methyl 4-bromo-3-oxobutyrate, from Penicillium citrinum IFO4631","volume":"71","author":"H Asako","year":"2005","journal-title":"Applied and environmental microbiology"},{"key":"pone.0260787.ref023","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s40643-018-0220-x","article-title":"Gene mining-based identification of aldo\u2013keto reductases for highly stereoselective reduction of bulky ketones","volume":"5","author":"C Liang","year":"2018","journal-title":"Bioresources and bioprocessing"},{"key":"pone.0260787.ref024","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.enzmictec.2015.06.004","article-title":"Cloning and characterization of a NADH-dependent aldo-keto reductase from a newly isolated Kluyveromyces lactis XP1461","volume":"77","author":"X Luo","year":"2015","journal-title":"Enzyme Microb Technol"},{"key":"pone.0260787.ref025","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.tibtech.2008.03.004","article-title":"New opportunities for biocatalysis: making pharmaceutical processes greener","volume":"26","author":"JM Woodley","year":"2008","journal-title":"Trends Biotechnol"},{"key":"pone.0260787.ref026","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.bej.2013.04.005","article-title":"Purification and characterization of fluorinated ketone reductase from Geotrichum candidum NBRC 5767","volume":"76","author":"C Cao","year":"2013","journal-title":"Biochemical engineering journal"},{"key":"pone.0260787.ref027","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1007\/s00253-004-1870-5","article-title":"Biotransformations for the production of the chiral drug (S)-Duloxetine catalyzed by a novel isolate of Candida tropicalis","volume":"67","author":"P Soni","year":"2005","journal-title":"Applied Microbiology & Biotechnology"},{"key":"pone.0260787.ref028","doi-asserted-by":"crossref","first-page":"W252","DOI":"10.1093\/nar\/gku340","article-title":"SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information","volume":"42","author":"M Biasini","year":"2014","journal-title":"Nucleic acids research"},{"key":"pone.0260787.ref029","doi-asserted-by":"crossref","first-page":"281","DOI":"10.2165\/11539240-000000000-00000","article-title":"Duloxetine","volume":"50","author":"MP Knadler","year":"2011","journal-title":"Clinical pharmacokinetics"},{"key":"pone.0260787.ref030","doi-asserted-by":"crossref","first-page":"1428","DOI":"10.1128\/aem.63.4.1428-1433.1997","article-title":"Organic solvent tolerance and antibiotic resistance increased by overexpression of marA in Escherichia coli","volume":"63","author":"H Asako","year":"1997","journal-title":"Applied and Environmental Microbiology"},{"key":"pone.0260787.ref031","doi-asserted-by":"crossref","first-page":"1951","DOI":"10.1021\/acssynbio.0c00267","article-title":"Lyophilized cell-free systems display tolerance to organic solvent exposure","volume":"9","author":"MS Lee","year":"2020","journal-title":"ACS Synthetic Biology"},{"key":"pone.0260787.ref032","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1007\/s12010-019-03091-1","article-title":"Coenzyme binding site analysis of an isopropanol dehydrogenase with wide substrate spectrum and excellent organic solvent tolerance","volume":"190","author":"W Jiang","year":"2020","journal-title":"Applied Biochemistry and Biotechnology"},{"key":"pone.0260787.ref033","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.jbiotec.2019.12.014","article-title":"Improvement in organic solvent resistance and activity of metalloprotease by directed evolution","volume":"309","author":"F Zhu","year":"2020","journal-title":"Journal of Biotechnology"},{"key":"pone.0260787.ref034","doi-asserted-by":"crossref","DOI":"10.1128\/AEM.00106-20","article-title":"Organic-solvent-tolerant carboxylic ester hydrolases for organic synthesis","volume":"86","author":"A Bollinger","year":"2020","journal-title":"Applied and Environmental Microbiology"},{"key":"pone.0260787.ref035","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1016\/j.ijbiomac.2020.05.021","article-title":"Characterization and efficient production of a thermostable, halostable and organic solvent-stable cellulase from an oil reservoir","volume":"159","author":"T Zhu","year":"2020","journal-title":"International journal of biological macromolecules"},{"key":"pone.0260787.ref036","first-page":"198","article-title":"Structure, function and biocatalytic applications of microbial aldo-keto reductases","volume":"046","author":"Lizhen Shi","year":"2017","journal-title":"Bulletin of Fermentation Science and Technology"},{"key":"pone.0260787.ref037","article-title":"Novel aldo-keto reductases for the biocatalytic conversion of 3-hydroxybutanal to 1, 3-butanediol: structural and biochemical studies","volume":"83","author":"T Kim","year":"2017","journal-title":"Applied and environmental microbiology"}],"container-title":["PLOS 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