Q Sepharose 강 음이온 교환막을 이용한 유청에 포함된 단백질 분리

최두영; 노경호; Du Young Choi; Kyung Ho Row

HWAHAK KONGHAK, Vol.40, No.4, 461-466, August, 2002

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Q Sepharose 강 음이온 교환막을 이용한 유청에 포함된 단백질 분리

Separation of Proteins in Whey by Strong Anion-Exchange Membrane of Q Sepharose

최두영, 노경호 ^{1, †}

인하대학교 초정밀분리기술센터, 인천 402-751
¹인하대학교 공과대학 화학공학과, 인천 402-751

Du Young Choi, and Kyung Ho Row^{1, †}

Center for Advanced Bioseparation Technology, Inha University, Inchon 402-751, Korea
¹Dept. of Chem. Eng., Inha University, Inchon 402-751, Korea

E-mail:

초록

선형 구배 용매 조성법을 이용하여 강 음이온 교환막으로 유청 단백질 중에 있는 α-lactalbumin, BSA, β-lactoglobulin A와 B를 분리하였다. 전처리 방법으로 사용한 원심분리형 UF 필터의 분자량 한계가 30,000인 막이 10,000에 비해 효과적이었다. NaCl의 농도는 유청 단백질 분리에 적합한 이동상의 조성에 맞게 변화되었다. 이동상은 buffer A(20 mM piperazine-HCl pH 6.4), buffer B(buffer A+1 M NaCl)를 사용하였다. 최적 이동상의 조성은 buffer A/buffer B(vol%)=95/5-90/10(0-5 min), 90/10(5-10 min), 90/10-70/30(10-45 min), 70/30(45-55 min), 70/30-10/90(55-60 min)이었다. 위의 조건으로 실험을 수행한 결과 4개의 주요 유청 단백질을 만족스럽게 분리하였다.

Strong anion-exchange membrane in the linear gradient elution method was experimented for the separation of the major whey proteins of α-lactalbumin, BSA, β-lactoglobulin(A, B). This centrifugal UF filters were used for the pretreatment of proteins. The membrane with nominal molecular weight limit 30,000 was more effective than that with 10,000. The salt concentrations were changed to determine the mobile phase composition for separating the whey proteins. The mobile phase was composed of buffer A(20 mM piperazine-HCl pH 6.4) and buffer B(buffer A+1 M NaCl). The optimum mobile phase composition was determined as buffer A/buffer B(vol%)=95/5-90/10(0-5 min), 90/10(5-10 min), 90/10-70/30(10-45 min), 70/30(45-55 min), 70/30-10/90(55-60 min). With this experimental condition of the linear-gradient elution mode, four major whey proteins were satisfactorily separated.

Keywords:Whey Proteins;Anion Exchange Membrane;Buffer;Mobile Phase Composition

[References]

Meisel H, Dtsch. Milchwirtsch, 31, 967 (1991)
Mclntosh GH, Regester GO, Le Leu RK, Royle PJ, Smithers GW, J. Nutr., 125, 809 (1995)
http://100.empas.com/entry.html/?i=150267&Ad=epson
Ye X, Yoshida S, Ng TB, Int. J. Biochem. Cell. Biology, 32, 1143 (2000)
Korhonen H, Pihlanto-Leppala A, Rantamaki P, Tupasela T, Trends Food Sci. Technol., 9, 307 (1998)
Bury D, Jelen P, Kimura K, Dairy J., 8, 149 (1998)
Lieske B, Konrad G, Dairy J., 6, 13 (1996)
Gerberding SJ, Byers CH, J. Chromatogr. A, 808, 141 (1998)
Splitt H, Mackenstedt I, Freitag R, J. Chromatogr. A, 729, 87 (1996)
Zydney AL, Dairy J., 8, 243 (1998)
Hahn R, Schulz PM, Schaupp C, Jungbauer A, J. Chromatogr. A, 795, 277 (1998)
Hutchens TW, Magnuson JS, Yip T, J. Immunol. Methods, 128, 98 (1990)
Strange ED, Malin EL, Van Hekken DL, Basch JJ, J. Chromatogr., 624, 81 (1992)
Alves JGLF, Chumpitaz LDA, da Silva LHM Franco TT, Meirelles AJA, J. Chromatogr. B, 743, 235 (2000)
Rito-Palomares M, Hernandez M, J. Chromatogr. B, 711, 81 (1998)
da Silva LHM, Meirelles AJA, Carbohydrate Polym., 42, 279 (2000)
Konrad G, Lieske B, Faber W, Int. Dairy J., 10, 713 (2000)
Lucas D, Rabiller-Baudry M, Millesime L, Chaufer B, Daufin G, J. Membr. Sci., 148(1), 1 (1998)
Punidadas P, Rizvi SSH, Food Res. Int., 31, 265 (1998)
Josic D, Buchacher A, Junhbauer A, J. Chromatogr. B, 752, 191 (2001)
Splitt H, Mackenstedt I, Freitag R, J. Chromatogr. A, 729, 87 (1996)
Manji B, Hill A, Kakuda Y, Irvine DM, J. Dairy Sci., 68, 3176 (1985)
Shan L, Anderson DJ, J. Chromatogr. A, 909, 191 (2001)
Keith Roper D, Lightfoot EN, J. Chromatogr. A, 702, 3 (1995)
Koo YM, Row KH, J. Korean Ind. Eng. Chem., 11(8), 807 (2000)
http://100.empas.com/entry.html/?i=52227&Ad=kfc

[Referenced By]

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[1] Meisel H, Dtsch. Milchwirtsch, 31, 967 (1991)

[2] Mclntosh GH, Regester GO, Le Leu RK, Royle PJ, Smithers GW, J. Nutr., 125, 809 (1995)

[3] http://100.empas.com/entry.html/?i=150267&Ad=epson

[4] Ye X, Yoshida S, Ng TB, Int. J. Biochem. Cell. Biology, 32, 1143 (2000)

[5] Korhonen H, Pihlanto-Leppala A, Rantamaki P, Tupasela T, Trends Food Sci. Technol., 9, 307 (1998)

[6] Bury D, Jelen P, Kimura K, Dairy J., 8, 149 (1998)

[7] Lieske B, Konrad G, Dairy J., 6, 13 (1996)

[8] Gerberding SJ, Byers CH, J. Chromatogr. A, 808, 141 (1998)

[9] Splitt H, Mackenstedt I, Freitag R, J. Chromatogr. A, 729, 87 (1996)

[10] Zydney AL, Dairy J., 8, 243 (1998)

[11] Hahn R, Schulz PM, Schaupp C, Jungbauer A, J. Chromatogr. A, 795, 277 (1998)

[12] Hutchens TW, Magnuson JS, Yip T, J. Immunol. Methods, 128, 98 (1990)

[13] Strange ED, Malin EL, Van Hekken DL, Basch JJ, J. Chromatogr., 624, 81 (1992)

[14] Alves JGLF, Chumpitaz LDA, da Silva LHM Franco TT, Meirelles AJA, J. Chromatogr. B, 743, 235 (2000)

[15] Rito-Palomares M, Hernandez M, J. Chromatogr. B, 711, 81 (1998)

[16] da Silva LHM, Meirelles AJA, Carbohydrate Polym., 42, 279 (2000)

[17] Konrad G, Lieske B, Faber W, Int. Dairy J., 10, 713 (2000)

[18] Lucas D, Rabiller-Baudry M, Millesime L, Chaufer B, Daufin G, J. Membr. Sci., 148(1), 1 (1998)

[19] Punidadas P, Rizvi SSH, Food Res. Int., 31, 265 (1998)

[20] Josic D, Buchacher A, Junhbauer A, J. Chromatogr. B, 752, 191 (2001)

[21] Splitt H, Mackenstedt I, Freitag R, J. Chromatogr. A, 729, 87 (1996)

[22] Manji B, Hill A, Kakuda Y, Irvine DM, J. Dairy Sci., 68, 3176 (1985)

[23] Shan L, Anderson DJ, J. Chromatogr. A, 909, 191 (2001)

[24] Keith Roper D, Lightfoot EN, J. Chromatogr. A, 702, 3 (1995)

[25] Koo YM, Row KH, J. Korean Ind. Eng. Chem., 11(8), 807 (2000)

[26] http://100.empas.com/entry.html/?i=52227&Ad=kfc