Stereoselective biotransformation of racemic mandelic acid using immobilized laccase and (S)-mandelate dehydrogenase

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 K_m 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.

Keywords

Biocatalysis / (S)-Mandelate dehydrogenase / Laccase / Immobilization / Chitosan / Bi-enzymatic system

References

[1] Baminger U, Ludwig R, Galhaup C, Leitner C, Kulbe K, Haltrich D. Continuous enzymatic regeneration of redox mediators used in biotransformation reactions employing flavoproteins. J Mol Catal B Enzym, 2001, 11: 541-550.

[2] Blank L, Ebert B, Buehler K, Bühler B. Redox biocatalysis and metabolism: molecular mechanisms and metabolic network analysis. Antioxid Redox Signal, 2010, 13: 349-394.

[3] Çetinus S, Öztop H. Immobilization of catalase into chemically crosslinked chitosan beads. Enzyme Microb Technol, 2003, 32: 889-894.

[4] Delanoy G, Li Q, Yu J. Activity and stability of laccase in conjugation with chitosan. Int J Biol Macromol, 2005, 35: 89-95.

[5] Dewanti A, Mitra B. A transient intermediate in the reaction catalyzed by (S)-mandelate dehydrogenase from Pseudomonas putida. Biochemistry, 2003, 42: 12893-12901.

[6] Gupta KC, Kumar MNVR. Preparation, characterization and release profiles of pH-sensitive chitosan beads. Polym Int, 2000, 49: 141-146.

[7] He Y, Xu J, Pan J, Ouyang L, Xu Y. Preparation of (R)-(-)-mandelic acid and its derivatives from racemates by enantioselective degradation with a newly isolated bacterial strain Alcaligenes sp. ECU0401. Bioprocess Biosyst Eng, 2008, 31: 445-451.

[8] Huang H, Xu J. Preparation of (S)-mandelic acid from racemate using growing cells of Pseudomonas putida ECU1009 with (R)-mandelate degradation activity. Biochem Eng J, 2006, 30: 11-15.

[9] Jiang B, Zhang Y. Immobilization of catalase on crosslinked polymeric hydrogels-effect of anion on the activity of immobilized enzyme. Eur Polym J, 1993, 29: 1251-1254.

[10] Jiang D, Long S, Huang J, Xiao H, Zhou J. Immobilization of Pycnoporus sanguineus laccase on magnetic chitosan microspheres. Biochem Eng J, 2005, 25: 15-23.

[11] Kaur I, Bhati P, Bala K, Kanwar S (2014) Effective immobilization of lipase onto a porous gelatin-co-poly(vinyl alcohol) copolymer and evaluation of its hydrolytic properties. J Appl Polym Sci 131

[12] Kurokawa J, Asano M, Nomoto S, Makino Y, Itoh N. Gene cloning and characterization of dihydrolipoamide dehydrogenase from Microbacterium luteolum: a useful enzymatic regeneration system of NAD(+) from NADH. J Biosci Bioeng, 2010, 109: 218-223.

[13] Lemainque A, Braun J, Le F. Influence of polymerization of the D-amino acid oxidase on the behavior of the enzyme covalently immobilized on chitosan. Eur J Biochem, 1988, 174: 171-176.

[14] Li Y, Huang X, Qu Y. A strategy for efficient immobilization of laccase and horseradish peroxidase on single-walled carbon nanotubes. J Chem Technol Biotechnol, 2013, 88: 2227-2232.

[15] Lorenz H, Sapoundjiev D, Seidel-Morgenstern A. Enantiomeric mandelic acid system-melting point phase diagram and solubility in water. J Chem Eng Data, 2002, 47: 1280-1284.

[16] Ludwig R, Ozga M, Zamocky M, Peterbauer C, Kulbe K, Haltrich D. Continuous enzymatic regeneration of electron acceptors used by flavoenzymes: cellobiose dehydrogenase-catalyzed production of lactobionic acid as an example. Biocatal Biotransform, 2004, 22: 97-104.

[17] Mao S, Zhang Y, Rohanl S, Ray AK. Chromatographic resolution and isotherm determination of (R, S)-mandelic acid on Chiralcel-OD column. J Sep Sci, 2012, 35: 2273-2281.

[18] Mohamed A, Naby A. Immobilization of Aspergillus niger NRC 107 xylanase and β-xylosidase, and properties of the immobilized enzymes. Appl Biochem Biotechnol, 1993, 38: 69-81.

[19] Reed M, Lieb A, Nijhout H. The biological significance of substrate inhibition: a mechanism with diverse functions. BioEssays, 2010, 32: 422-429.

[20] Rodrigues R, Ortiz C, Berenguer-Murcia Á, Torres R, Fernández-Lafuente R. Modifying enzyme activity and selectivity by immobilization. Chem Soc Rev, 2013, 42: 6290-6307.

[21] Silva J, Macedo G, Rodrigues D. Immobilization of Candida antarctica lipase B by covalent attachment on chitosan-based hydrogels using different support activation strategies. Biochem Eng J, 2012, 60: 16-24.

[22] Takahashi E, Nakamichi K, Furui M. R-(-)-Mandelic acid production from racemic mandelic acids using Pseudomonas polycolor IFO 3918 and Micrococcus freudenreichii FERM-P 13221. J Ferment Bioeng, 1995, 80: 247-250.

[23] Thurston C. The structure and function of fungal laccases. Microbiology, 1994, 140: 19-26.

[24] Tulashie S, Lorenz H, Seidel-Morgenstern A. Solubility of mandelic acid enantiomers and their mixtures in three chiral solvents. J Chem Eng Data, 2010, 55: 5196-5200.

[25] Wang P, Yang J, Jiang L, Feng J, Yang C, Li D. A bi-enzymatic system for efficient enantioselective bioconversion of racemic mandelic acid. J Mol Catal B Enzym, 2013, 94: 47-50.

[26] Wang P, Li D, Yang J, Jiang L, Feng J, Yang C, Shi R. Immobilization of (S)-mandelate dehydrogenase and its catalytic performance on stereoselective transformation of mandelic acid. J Taiwan Inst Chem Eng, 2014, 45: 744-748.

[27] Zhang Z, Xu J, He Y, Ouyang L, Liu Y, Imanaka T. Efficient production of (R)-(-)-mandelic acid with highly substrate/product tolerant and enantioselective nitrilase of recombinant Alcaligenes sp. Process Biochem, 2010, 45: 887-891.

[28] Zhang Z, Pan J, Li C, Yu H, Zheng G, Ju X, Xu J. Efficient production of (R)-(-)-mandelic acid using glutaraldehyde cross-linked Escherichia coli cells expressing Alcaligenes sp. nitrilase. Bioprocess Biosyst Eng, 2014, 37: 1241-1248.