Expression of soluble Saccharomyces cerevisiae alcohol dehydrogenase in Escherichia coli applicable to oxido-reduction bioconversions

Utekal, Pavol; Tóth, Csaba; Illésová, Anikó; Kois, Pavol; Bocanova, Lucia; Turňa, Ján; Drahovská, Hana; Stuchlı́k, Stanislav

doi:10.2478/s11756-014-0376-6

Cited by 6 publications

(2 citation statements)

References 26 publications

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“…The LB medium differed from the other two in that it lacked glycerol and had a unique salt content. Ideal results were obtained with the BL21(DE3) and LEMO21(DE3) strains grown in TB medium at 28°C as previously described (Utekal et al, 2014 ; Levarski et al, 2018 ). Maximum RrADH expression with respect to soluble fraction content was detected 18 h after induction in LEMO21(DE3) ( Figure 2 ).…”

Section: Discussionsupporting

confidence: 62%

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Recombinant Enzymatic Redox Systems for Preparation of Aroma Compounds by Biotransformation

et al. 2021

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The aim of this study was to develop immobilized enzyme systems that reduce carbonyl compounds to their corresponding alcohols. The demand for natural aromas and food additives has been constantly growing in recent years. However, it can no longer be met by extraction and isolation from natural materials. One way to increase the availability of natural aromas is to prepare them by the enzymatic transformation of suitable precursors. Recombinant enzymes are currently being used for this purpose. We investigated trans-2-hexenal bioreduction by recombinant Saccharomyces cerevisiae alcohol dehydrogenase (ScADH1) with simultaneous NADH regeneration by recombinant Candida boidinii formate dehydrogenase (FDH). In a laboratory bioreactor with two immobilized enzymes, 88% of the trans-2-hexenal was transformed to trans-2-hexenol. The initial substrate concentration was 3.7 mM. The aldehyde destabilized ScADH1 by eluting Zn2+ ions from the enzyme. A fed-batch operation was used and the trans-2-hexenal concentration was maintained at a low level to limit the negative effect of Zn2+ ion elution from the immobilized ScADH1. Another immobilized two-enzyme system was used to reduce acetophenone to (S)-1-phenylethanol. To this end, the recombinant alcohol dehydrogenase (RrADH) from Rhodococcus ruber was used. This biocatalytic system converted 61% of the acetophenone to (S)-1-phenylethanol. The initial substrate concentration was 8.3 mM. All enzymes were immobilized by poly-His tag to Ni2+, which formed strong but reversible bonds that enabled carrier reuse after the loss of enzyme activity.

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Section: Discussionsupporting

confidence: 62%

“…All bacterial strains and plasmids were used in ADH1 production from Saccharomyces cerevisiae (Utekal et al, 2014 ; Levarski et al, 2018 ) and in FHD production from Candida boidinii . Gene expressions and enzyme preparations were described in a previous report (Levarski et al, 2018 ).…”

Section: Methodsmentioning

confidence: 99%

Recombinant Enzymatic Redox Systems for Preparation of Aroma Compounds by Biotransformation

et al. 2021

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Production of N-glycosylated alcohol dehydrogenase in Escherichia coli

Levarski,

Bírová,

Hriňová

et al. 2024

Biologia

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N-glycosylation of recombinant proteins using bacterial glycosylation system has proven to be a valuable although developing tool ultimately applicable to various industries. When used for enzyme engineering, it offers the possibility of increased stability or immobilization route and thus increasing effectiveness of e.g. biotransformation or other biocatalysis procedures. One such promising enzyme is alcohol dehydrogenase (ADH) for use in redox biotransformation reactions. Given the current possibilities of recombinant enzyme production, including major advances in glycoengineering and glycoprotein production in bacterial organisms, the aim of this work was the production of thermotolerant ADH from Rhodococcus ruber (RrADH) in glycosylated form in Escherichia coli. We have successfully developed a dual plasmid expression system enabling glycosylation of target proteins utilizing a glyco-tag approach. We were able to produce RrADH in soluble form and at the same time we detected a bacterial glycan conjugated to RrADH as well as the activity of the enzyme. The glycan bound to recombinant enzyme can be used for oriented covalent immobilization of the enzyme, which would increase the potential for its practical application in biotransformation of various compounds.

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Engineering of Saccharomyces cerevisiae for enhanced production of L-lactic acid by co-expression of acid-stable glycolytic enzymes from Picrophilus torridus

Ryu

Kim

Yang

et al. 2018

Korean J. Chem. Eng.

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Expression of soluble Saccharomyces cerevisiae alcohol dehydrogenase in Escherichia coli applicable to oxido-reduction bioconversions

Cited by 6 publications

References 26 publications

Recombinant Enzymatic Redox Systems for Preparation of Aroma Compounds by Biotransformation

Recombinant Enzymatic Redox Systems for Preparation of Aroma Compounds by Biotransformation

Production of N-glycosylated alcohol dehydrogenase in Escherichia coli

Engineering of Saccharomyces cerevisiae for enhanced production of L-lactic acid by co-expression of acid-stable glycolytic enzymes from Picrophilus torridus

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