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Selective Enzymatic Transformation to Aldehydes in vivo by Fungal Carboxylate Reductase from Neurospora crassa

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논문

Selective Enzymatic Transformation to Aldehydes in vivo by Fungal Carboxylate Reductase from Neurospora crassa

학술지

Advanced synthesis & catalysis

저자명

Schwendenwein, Daniel; Fiume, Giuseppe; Weber, Hansjö rg; Rudroff, Florian; Winkler, Margit

초록

<P><B>Abstract</B></P><P>The enzymatic reduction of carboxylic acids is in its infancy with only a handful of biocatalysts available to this end. We have increased the spectrum of carboxylate&#x2010;reducing enzymes (CARs) with the sequence of a fungal CAR from <I>Neurospora crassa</I> OR74A (<I>Nc</I>CAR). <I>Nc</I>CAR was efficiently expressed in <I>E. coli</I> using an autoinduction protocol at low temperature. It was purified and characterized <I>in vitro</I>, revealing a broad substrate acceptance, a pH optimum at pH 5.5&#x2013;6.0, a <I>T</I><SUB>m</SUB> of 45 °C and inhibition by the co&#x2010;product pyrophosphate which can be alleviated by the addition of pyrophosphatase. The synthetic utility of <I>Nc</I>CAR was demonstrated in a whole&#x2010;cell biotransformation using the <I>Escherichia coli</I> K&#x2010;12 MG1655 RARE strain in order to suppress overreduction to undesired alcohol. The fragrance compound piperonal was prepared from piperonylic acid (30 mM) on gram scale in 92 % isolated yield in >98% purity. This corresponds to a productivity of 1.5 g/L/h.</P><P/>

발행연도

2016

발행기관

John Wiley and Sons Inc.

ISSN

1615-4150

ISSN

1615-4169

358

21

페이지

pp.3414-3421

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논문; 2016-12-31

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