Exploiting the enzymatic machinery of Arthrobacter atrocyaneus for oxidative kinetic resolution of secondary alcohols

Silva, Camila R.; Souza, Juliana C.; Araújo, Lidiane S.; Kagohara, Edna; Garcia, Thaís P.; Pelizzari, Vivian H.; Andrade, Leandro H.

doi:10.1016/j.molcatb.2012.06.013

Cited by 9 publications

(9 citation statements)

References 31 publications

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“…[19] Oxidative kinetic resolutions have in most cases been applied to racemic alcohols. [20][21][22] Conventionally,k inetic resolutions are performed with hydrolases, [23][24][25][26][27][28] because of their simple handling without the need for cofactor regeneration systems. In addition to these, the described kinetic resolution of the hydroxy ketone rac-1 (Scheme 1) represents am eans of biocatalytic access to enantiomerically pure tertiary alcohols throught he use of dehydrogenases.…”

Section: Discussionmentioning

confidence: 99%

Screening, Molecular Cloning, and Biochemical Characterization of an Alcohol Dehydrogenase from Pichia pastoris Useful for the Kinetic Resolution of a Racemic β‐Hydroxy‐β‐trifluoromethyl Ketone

et al. 2016

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The stereoselective synthesis of chiral 1,3-diols with the aid of biocatalysts is an attractive tool in organic chemistry. Besides the reduction of diketones, an alternative approach consists of the stereoselective reduction of β-hydroxy ketones (aldols). Thus, we screened for an alcohol dehydrogenase (ADH) that would selectively reduce a β-hydroxy-β-trifluoromethyl ketone. One potential starting material for this process is readily available by aldol addition of acetone to 2,2,2-trifluoroacetophenone. Over 200 strains were screened, and only a few yeast strains showed stereoselective reduction activities. The enzyme responsible for the reduction of the β-hydroxy-β-trifluoromethyl ketone was identified after purification and subsequent MALDI-TOF mass spectrometric analysis. As a result, a new NADP(+) -dependent ADH from Pichia pastoris (PPADH) was identified and confirmed to be capable of stereospecific and diastereoselective reduction of the β-hydroxy-β-trifluoromethyl ketone to its corresponding 1,3-diol. The gene encoding PPADH was cloned and heterologously expressed in Escherichia coli BL21(DE3). To determine the influence of an N- or C-terminal His-tag fusion, three different recombinant plasmids were constructed. Interestingly, the variant with the N-terminal His-tag showed the highest activity; consequently, this variant was purified and characterized. Kinetic parameters and the dependency of activity on pH and temperature were determined. PPADH shows a substrate preference for the reduction of linear and branched aliphatic aldehydes. Surprisingly, the enzyme shows no comparable activity towards ketones other than the β-hydroxy-β-trifluoromethyl ketone.

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Section: Discussionmentioning

confidence: 99%

Screening, Molecular Cloning, and Biochemical Characterization of an Alcohol Dehydrogenase from Pichia pastoris Useful for the Kinetic Resolution of a Racemic β‐Hydroxy‐β‐trifluoromethyl Ketone

et al. 2016

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“…Finally, the ethyl acetate layer was directly subjected to GC analysis on a Chirasil-Dex CB column (25 m × 0.32 mm; 0.25 µm film; Varian). The GC settings were as follows: injector 220 °C; 1.0 bar constant pressure H 2 flow; temperature program: initial temperature 60 °C, 85 °C/rate 1.5 °C per min, hold 3 min; The absolute configuration of 2-octanol was assigned by comparison of the elution order on chiral GC with literature known data [ 28 ] and by derivatization of commercial ( R )- and ( S )-2-octanol. Retention times were 8.0 min for 2-octanone, 13.9 min for ( S )-octan-2-yl acetate and 17.2 min for ( R )-octan-2-yl acetate.…”

Section: Methodsmentioning

confidence: 99%

Enantiocomplementary Yarrowia lipolytica Oxidoreductases: Alcohol Dehydrogenase 2 and Short Chain Dehydrogenase/Reductase

et al. 2013

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Enzymes of the non-conventional yeast Yarrowia lipolytica seem to be tailor-made for the conversion of lipophilic substrates. Herein, we cloned and overexpressed the Zn-dependent alcohol dehydrogenase ADH2 from Yarrowia lipolytica in Escherichia coli. The purified enzyme was characterized in vitro. The substrate scope for YlADH2 mediated oxidation and reduction was investigated spectrophotometrically and the enzyme showed a broader substrate range than its homolog from Saccharomyces cerevisiae. A preference for secondary compared to primary alcohols in oxidation direction was observed for YlADH2. 2-Octanone was investigated in reduction mode in detail. Remarkably, YlADH2 displays perfect (S)-selectivity and together with a highly (R)-selective short chain dehydrogenase/ reductase from Yarrowia lipolytica it is possible to access both enantiomers of 2-octanol in >99% ee with Yarrowia lipolytica oxidoreductases.

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“…Álcoois secundários enantiomericamente puros são importantes blocos construtores bem como auxiliares quirais para a indústria farmacêutica, agroquímica e química fina. 12 Diversas estratégias são empregadas para o preparo desses compostos, como por exemplo a hidrogenação de cetonas pró-quirais catalisada por complexos metálicos, 13,14,15 a redução de cetonas pró-quirais por biocatalisadores, 16,17,18,19 a resolução cinética dinâmica, 20,21 a resolução cinética oxidativa, 22 desracemização de alcoóis secundários, 23,24,25,26 processos enantioconvergentes, 27 entre outros. O 1-feniletanol e seus derivados contendo substituintes no anel aromático tem sido utilizados como substratos modelo em processos biocatalícos.…”

Section: Obtenção De áLcoois Secundários Quiraisunclassified

“…O 1-feniletanol e seus derivados contendo substituintes no anel aromático tem sido utilizados como substratos modelo em processos biocatalícos. 22 Em particular, o 1-(3-hidroxifenil)-etanol (1) é um composto de interesse sintético pois na sua forma quiral, (R)-1, é matéria prima no preparo da Rivastigmina, um medicamento utilizado no tratamento do mal de Alzheimer. 28 O álcool quiral (R)-1 foi obtido a partir da redução da 3-hidroxiacetofenona por meio da utilização de um complexo metálico de irídio.…”

Section: Obtenção De áLcoois Secundários Quiraisunclassified

“…32 Bosetti, A.; Bianchi, D.; Cesti, P.; Golini, P.; Spezia, S. Journal of the Chemical Society, Perkin Transactions 1 1992, 2395. 33 22 Diferentes substituintes no anel aromático foram testados, nas posições orto, meta e para. Foram obtidos os álcoois de configuração absoluta S, com ee>99% para a maioria dos substratos testados, evidenciando as vantagens de processos biocatalíticos em relação aos processos químicos convencionais.…”

Section: Obtenção De áLcoois Secundários Quiraisunclassified

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Desracemização de álcoois secundários por estereoinversão e redução de metilenocetonas por micro/organismos

Cazetta¹

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Em primeiro lugar agradeço a Deus pela vida e pelas oportunidades que me foram dadas.Em especial ao prof. José Augusto pela orientação, incentivo e exemplo profissional nesses longos anos nos quais trabalhamos juntos.Aos meus pais, Nice e Luiz Cazetta por me incentivarem a recomeçar todas as vezes que desanimei, por vibrarem comigo nos momentos de sucesso, por me apoiarem nas dificuldades e principalmente pelo exemplo.À Ticiane Liberato, que durante esses quatro anos passou de amiga para namorada, e então para esposa: pelo apoio, incentivo, paciência e dedicação. Pelo amor que me ajuda a ser uma pessoa melhor. Às minhas irmãs Eliana, Paula e Luiza pela cumplicidade, e aos meus cunhados Josa e Hugo pela amizade e incentivo. Aos meus sobrinhos Pedro, Matheus e Jonatas e sobrinhas Ana Pietra, Isabelle e Daphne pela alegria dos momentos juntos e pelas lições ensinadas. À comunidade do IQ, professores e funcionários. Em especial a Leandro Vilela pela amizade além do apoio profissional.

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Exploiting the enzymatic machinery of Arthrobacter atrocyaneus for oxidative kinetic resolution of secondary alcohols

Cited by 9 publications

References 31 publications

Screening, Molecular Cloning, and Biochemical Characterization of an Alcohol Dehydrogenase from Pichia pastoris Useful for the Kinetic Resolution of a Racemic β‐Hydroxy‐β‐trifluoromethyl Ketone

Screening, Molecular Cloning, and Biochemical Characterization of an Alcohol Dehydrogenase from Pichia pastoris Useful for the Kinetic Resolution of a Racemic β‐Hydroxy‐β‐trifluoromethyl Ketone

Enantiocomplementary Yarrowia lipolytica Oxidoreductases: Alcohol Dehydrogenase 2 and Short Chain Dehydrogenase/Reductase

Desracemização de álcoois secundários por estereoinversão e redução de metilenocetonas por micro/organismos

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