OBJECTIVES: (S)-Mandelate dehydrogenase (SMDH) and laccase were immobilized on chitosan. The bi-enzymatic system with immobilized SMDH and immobilized laccase was taken to catalyze the stereoselective transformation of racemic mandelic acid and (R)-mandelic acid was obtained from its racemic mixture. RESULTS: Characteristics of the immobilized enzymes were valuated. The optimum pH and temperature of the immobilized SMDH were found to be pH 3.4 and 45 °C, and these of the immobilized laccase were about pH 6.0 and 55 °C, respectively. The Km value of the immobilized SMDH for racemic mandelic acid was 0.27 mM and that of the immobilized laccase for ferrocyanide was 0.99 mM. The thermal and storage stabilities of these enzymes were improved with immobilization. The enantiomeric purity of the bi-enzymatically produced (R)-mandelic acid was determined to be over 99%. CONCLUSION: The immobilized bi-enzymatic system for the stereoselective transformation of racemic mandelic acid showed higher productivity, faster reaction velocity, and more stable catalytic ability.Graphical abstract.
OBJECTIVES: (S)-Mandelate dehydrogenase (SMDH) and laccase were immobilized on chitosan. The bi-enzymatic system with immobilized SMDH and immobilized laccase was taken to catalyze the stereoselective transformation of racemic mandelic acid and (R)-mandelic acid was obtained from its racemic mixture. RESULTS: Characteristics of the immobilized enzymes were valuated. The optimum pH and temperature of the immobilized SMDH were found to be pH 3.4 and 45 °C, and these of the immobilized laccase were about pH 6.0 and 55 °C, respectively. The Km value of the immobilized SMDH for racemic mandelic acid was 0.27 mM and that of the immobilized laccase for ferrocyanide was 0.99 mM. The thermal and storage stabilities of these enzymes were improved with immobilization. The enantiomeric purity of the bi-enzymatically produced (R)-mandelic acid was determined to be over 99%. CONCLUSION: The immobilized bi-enzymatic system for the stereoselective transformation of racemic mandelic acid showed higher productivity, faster reaction velocity, and more stable catalytic ability.Graphical abstract.