Selective oxidation and reduction reactions with cofactor regeneration mediated by galactitol-, lactate-, and formate dehydrogenases immobilized on magnetic nanoparticles


DEMİR A. G. , Talpur F. N. , Sopaci S. B. , Kohring G., ÇELİK A.

JOURNAL OF BIOTECHNOLOGY, cilt.152, ss.176-183, 2011 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 152 Konu: 4
  • Basım Tarihi: 2011
  • Doi Numarası: 10.1016/j.jbiotec.2011.03.002
  • Dergi Adı: JOURNAL OF BIOTECHNOLOGY
  • Sayfa Sayıları: ss.176-183

Özet

Rapid immobilization with the one-pot purification of galactitol dehydrogenase (GatDH) and formate dehydrogenase (FDH) is achieved by using iminodiacetic acid (IDA) with chelated Co2+ modified magnetic nanoparticles as a carrier. Lactate dehydrogenase (LDH) from recombinant Escherichia coli and FDH commencing Candida methylica were used as an auxiliary enzyme for the regeneration of NADH/NAD(+) with a representative synthesis of (S)-1,2-propanediol and L-tagatose starting from hydroxyacetone and galactitol. The affinity magnetic nanoparticles were characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR), while the purity of GatDH and FDH was assayed by SDS-PAGE analysis. The immobilized two-enzyme system, reflecting the pH dependence of its constituent enzymes, showed optimal activity at pH 7 and 8 for (S)-1,2-propanediol and L-tagatose production, respectively. The immobilized enzyme system retained up to 70% of its activity after one week of repeated use. The use of affinity magnetic nanoparticles offers the advantage of a one-pot purification of His(6)-tagged GatDH and FDH followed by the production of rare sugar and chiral diol. (C) 2011 Elsevier B.V. All rights reserved.