Purification of Native and Modified Enzymes Using a Reactive Aqueous Two-Phase System

Yeounseoo Kim; Kwinam Park; Hyejin Lee; Seokheung Jang; Ho-Cheol Song; Ho-Chul Shin; Jong Jin Park; Jinwon Park; Sanjeev Maken

Journal of Industrial and Engineering Chemistry, Vol.10, No.3, 384-388, May, 2004

Yeounseoo Kim, Kwinam Park, Hyejin Lee, Seokheung Jang, Ho-Cheol Song, Ho-Chul Shin, Jong Jin Park, Jinwon Park^†, and Sanjeev Maken¹

Department of Chemical Engineering, Yonsei University, Seoul 120-749, Korea
¹Department of Applied Sciences, College of Engineering, C.R. State College, Murthal-139039, Hargana, India

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We describe the purification of native and modified enzymes using a reactive aqueous two-phase systems. The two-phase systems consist of a hydrophilic AKM copolymer and dextran. Cellulase was modified with the AKM copolymer; the degree of modification is directly proportional to the number of maleic acid anhydride groups present in the AKM copolymer. Partitioning of the native and modified cellulases was also investigated in the aqueous two-phase AKM copolymer/dextran systems; the partition coefficient increases as the degree of modification in creases. The AKM-0531 copolymer is enzyme selective; the modified enzyme always shows separation that is better than that of the native enzyme.

Keywords:aqueous two-phase system;partitioning;enzyme;copolymers

[References]

Simonet F, Garnier C, Doublier JL, Food Hydrocolloids, 14, 591 (2000)
Piculell L, Bergfeldt K, Nilsson S, Bio-polymer Mixtures, S.E. Harding, S.E. Hill, and J.R. Mitchell, Eds., pp. 13-35, Nottingham University Press, Nottingham (1995)
Albertsson PA, Partition of Cell Particles and Macromolecules, 3rd Edn., John Wiley, New York (1986)
Asenjo JA, Mistry SL, Andrews BA, Merchuk JC, Biotechnol. Bioeng., 79(2), 217 (2002)
Hart RA, Lester PM, Reifsnyder DH, Ogez JR, Builder S, Bio-Technol., 12, 1113 (1994)
Johansson HO, Persson J, Tjerneld F, Biotechnol. Bioeng., 66(4), 247 (1999)
Park JW, Park KN, Song HC, Shin HC, J. Biotechnol., 93, 203 (2002)
Moon HY, Kim SY, Park JW, Kajiuchi T, Korean J. Chem. Eng., 10(4), 235 (1993)
Park JW, Korean J. Chem. Eng., 12(5), 523 (1995)
Persson J, Kaul A, Tjerneld F, J. Chromatogr. B, 743, 115 (2000)
Park KN, Park JW, J. Appl. Polym. Sci., 77(2), 368 (2000)
Habeeb AFSF, Anal. Biochem., 14, 328 (1966)
Miller GL, Anal. Chem., 31, 426 (1959)
Diamond AD, Hsu JT, Adv. Biochem. Eng. Biotech., 47, 89 (1992)

[Referenced By]

[Related Articles]

[1] Simonet F, Garnier C, Doublier JL, Food Hydrocolloids, 14, 591 (2000)

[2] Piculell L, Bergfeldt K, Nilsson S, Bio-polymer Mixtures, S.E. Harding, S.E. Hill, and J.R. Mitchell, Eds., pp. 13-35, Nottingham University Press, Nottingham (1995)

[3] Albertsson PA, Partition of Cell Particles and Macromolecules, 3rd Edn., John Wiley, New York (1986)

[4] Asenjo JA, Mistry SL, Andrews BA, Merchuk JC, Biotechnol. Bioeng., 79(2), 217 (2002)

[5] Hart RA, Lester PM, Reifsnyder DH, Ogez JR, Builder S, Bio-Technol., 12, 1113 (1994)

[6] Johansson HO, Persson J, Tjerneld F, Biotechnol. Bioeng., 66(4), 247 (1999)

[7] Park JW, Park KN, Song HC, Shin HC, J. Biotechnol., 93, 203 (2002)

[8] Moon HY, Kim SY, Park JW, Kajiuchi T, Korean J. Chem. Eng., 10(4), 235 (1993)

[9] Park JW, Korean J. Chem. Eng., 12(5), 523 (1995)

[10] Persson J, Kaul A, Tjerneld F, J. Chromatogr. B, 743, 115 (2000)

[11] Park KN, Park JW, J. Appl. Polym. Sci., 77(2), 368 (2000)

[12] Habeeb AFSF, Anal. Biochem., 14, 328 (1966)

[13] Miller GL, Anal. Chem., 31, 426 (1959)

[14] Diamond AD, Hsu JT, Adv. Biochem. Eng. Biotech., 47, 89 (1992)