• Title/Summary/Keyword: benzoylformate reductase

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Production of (R)-(-)-mandelic acid by electrochemically driven enzyme bioreactor

  • Kim, Mi-Hae;Yun, Se-Eok
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.642-645
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    • 2001
  • Enterococcus faecalis was cultivated under oxidative conditions established by adding some oxidants. FAD and lipoic acid either stimulate the biosynthesis of benzoylformate reductase or stabilize the enzyme, while $MV^{2+}$ enhance the biosynthesis of the oxidoreductase but destabilize it. Since $MV^{2+}$ destabilize the benzoylformate reductase, substituting FAD for $MV^{2+}$ as a redox mediator would be desirable. Production of (R)-(-)-mandelic acid by a coupled reaction between the enzymatic reaction using benzoylformate reductase and the electrocatalytic reduction under the conditions of 1.5 U LiDH $ml^{-1}$, 0.2 mM FAD, and 0.3 mM $NAD^+$ is now performing.

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Enhanced Production of Benzoylformate Reductase in Enterococcus faecalis under Oxidative Stress Established by Natural Electron Carriers

  • Baik, Sang-Ho;Cho, Pan-Ki;Kim, Mee-Hae;Yun, Sei-Eok
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.104-109
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    • 2003
  • Enhancement of the production of benzoylformate reductase (BFR) was attempted under oxidative stress established by natural electron carriers. -lipoic acid (LA), flavin adenine dinucleotide (FAD), and ubiquinone (UQ) did not inhibit growth of E. faecalis when their concentrations were as high as $10{\mu}M$, while $H_2O_2$ and methyl viologen ($MV^2+$) inhibited the bacterial growth. BFR activity in the bacterial extract had increased rapidly after 1 h of cultivation after the addition of $4{\mu}M$ of natural electron carriers, and the activity was maintained during further cultivation. BFR activity of the cells treated with the natural electron carriers was $40\%$ higher than that of the control. In the presence of $4{\mu}M\;H_2O_2\;and\;MV^2+$, BFR activity increased, reaching the highest activity at about 5 h cultivation, and then decreased with further cultivation. It seems that natural electron carriers not only stimulate the induction of BFR, but also stabilize the enzyme. BFR was hardly affected by LA, FAD, and UQ, while $H_2O_2\;and\;MV^2+$ inactivated the crude enzyme. The decrease of BFR activity in the presence of $H_2O_2\;and\;MV^2+$ might be ascribed to inactivation of the enzyme by the oxidants.