Novel Supports for Enzyme Immobilization Based on Surface-Grafted Copolymers of Polystyrene and Poly(ethylene glycol)

Jang-Woong Byun; Yoon-Sik Lee

Journal of Industrial and Engineering Chemistry, Vol.10, No.2, 283-289, March, 2004

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Novel Supports for Enzyme Immobilization Based on Surface-Grafted Copolymers of Polystyrene and Poly(ethylene glycol)

Jang-Woong Byun, and Yoon-Sik Lee^†

School of Chemical Engineering, Seoul National University, Seoul 151-744, Korea

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We have developed a useful method for preparing surface-grafted polystyrene (PS) beads with hydrophilic poly(ethylene glycol) (PEG) chains that uses simple ozone oxidation. To accomplish this goal, geltype PS beads (37-73 μm) were treated with moisture-saturated ozone gas to introduce hydroperoxide groups. By using these hydroperoxide groups as initiators, methacrylate-type PEG macromers were grafted onto the surface of the PS beads. The resulting PS-sg-PEG beads contained about 0.2-0.3 mmol/g of the PEG macromers and had diameters of 50-80 μm. We investigated the use of the PS-sg-PEG beads as supports for enzyme immobilization. Following the coupling of cyanuric chloride to the terminal hydroxyl groups, two kinds of protease were covalently irnmobilized. We found that a large amount of subtilisin (450-500 mg enzyme/g beads) could be loaded onto the beads without any significant loss of enzyme activity. In addition, the immobilized α-chymotrypsm could be used to synthesize dipeptide products in 45-60% yield in acetonitrile.

Keywords:ozone oxidation;surface modification;PS-sg-PEG beads;enzyme immobilization;peptide synthesis

[References]

Katzir EK, Kreamer DM, J. Mol. Catal. B-Enzym., 10, 157 (2000)
Ryu JC, Ryu TG, Yoo SJ, Hwang JY, Kim YH, Yang HS, J. Ind. Eng. Chem., 9(6), 735 (2003)
Blanco RM, Alvaro G, Guisan JM, Enzyme Microb. Technol., 13, 573 (1991)
Heras A, Acosta N, Biocatalysis, 11, 305 (1994)
Lozano P, Diego T, Iborra JL, Biotechnol. Lett., 17(6), 603 (1995)
Mitz M, Summaria L, Nature, 189, 576 (1961)
Hasine GH, Wang CC, J. Appl. Polym. Sci., 40, 255 (1990)
Gombotz WR, Guanghui W, Horbett TA, Hoffman AS, J. Biomed. Mater. Res., 25, 1547 (1991)
Jeon SL, Lee JH, Andrnde JD, J. Colloid Interface Sci., 142, 149 (1991)
Tsuchida E, Nishide H, Shimidazu N, Yamada A, Keneko M, Makromol. Chem. Rapid Commun., 2, 621 (1981)
Becker H, Lucas HW, Maul J, Pillai VNR, Anzinger H, Mutter M, Makromol. Chem. Rapid Commun., 3, 217 (1982)
Hellermann H, Lucas HW, Maul J, Pillai VNR, Mutter M, Makromol. Chem., 184, 2603 (1983)
Bayer E, Dengler M, Hemmasi B, Int. J. Pept. Protein Res., 25, 178 (1985)
Lee YS, Park BD, Lee HI, U.S. Patent, 5,466,758 (1995)
Gooding W, Baudart S, Deegan TL, Heisler K, Labadie JW, Newcomb WS, Porco JA, vanEikeren P, J. Comput. Chem., 1, 113 (1999)
Hubbell JA, Trends Polym. Sci., 2, 20 (1994)
Athawle VD, Rathi SC, J. Polym. Mater., 13, 335 (1996)
Finlay TH, Troll V, Levy M, Johnson AJ, Hodgins LT, Anal. Biochem., 87, 77 (1978)
Zaks A, Klibanov AM, J. Am. Chem. Soc., 119, 707 (1986)

[Referenced By]

[Related Articles]

[1] Katzir EK, Kreamer DM, J. Mol. Catal. B-Enzym., 10, 157 (2000)

[2] Ryu JC, Ryu TG, Yoo SJ, Hwang JY, Kim YH, Yang HS, J. Ind. Eng. Chem., 9(6), 735 (2003)

[3] Blanco RM, Alvaro G, Guisan JM, Enzyme Microb. Technol., 13, 573 (1991)

[4] Heras A, Acosta N, Biocatalysis, 11, 305 (1994)

[5] Lozano P, Diego T, Iborra JL, Biotechnol. Lett., 17(6), 603 (1995)

[6] Mitz M, Summaria L, Nature, 189, 576 (1961)

[7] Hasine GH, Wang CC, J. Appl. Polym. Sci., 40, 255 (1990)

[8] Gombotz WR, Guanghui W, Horbett TA, Hoffman AS, J. Biomed. Mater. Res., 25, 1547 (1991)

[9] Jeon SL, Lee JH, Andrnde JD, J. Colloid Interface Sci., 142, 149 (1991)

[10] Tsuchida E, Nishide H, Shimidazu N, Yamada A, Keneko M, Makromol. Chem. Rapid Commun., 2, 621 (1981)

[11] Becker H, Lucas HW, Maul J, Pillai VNR, Anzinger H, Mutter M, Makromol. Chem. Rapid Commun., 3, 217 (1982)

[12] Hellermann H, Lucas HW, Maul J, Pillai VNR, Mutter M, Makromol. Chem., 184, 2603 (1983)

[13] Bayer E, Dengler M, Hemmasi B, Int. J. Pept. Protein Res., 25, 178 (1985)

[14] Lee YS, Park BD, Lee HI, U.S. Patent, 5,466,758 (1995)

[15] Gooding W, Baudart S, Deegan TL, Heisler K, Labadie JW, Newcomb WS, Porco JA, vanEikeren P, J. Comput. Chem., 1, 113 (1999)

[16] Hubbell JA, Trends Polym. Sci., 2, 20 (1994)

[17] Athawle VD, Rathi SC, J. Polym. Mater., 13, 335 (1996)

[18] Finlay TH, Troll V, Levy M, Johnson AJ, Hodgins LT, Anal. Biochem., 87, 77 (1978)

[19] Zaks A, Klibanov AM, J. Am. Chem. Soc., 119, 707 (1986)