고정화 효소에 의한 섬유소 가수분해

오경근; 김승욱; 홍석인

HWAHAK KONGHAK, Vol.30, No.4, 467-472, August, 1992

Export Citation

고정화 효소에 의한 섬유소 가수분해

Cellulose Hydrolysis by Immobilized Enzyme

오경근, 김승욱, 홍석인

초록

Trichoderma reesei QM 9414로부터 생산된 셀룰라이제를 Ca-alginate로 고정화하여 안정도를 높임으로써 셀룰로오스를 가수분해하는데 장기간 사용할 수 있도록 하였다. 고정화 효소의 역가는 Na-alginate 와 CaCl₂ 용액의 농도가 각각 3%(w/v), 0.4M일 때 가장 좋은 결과를 보여준다. 고정화 겔 비드의 강도는 Na-alginate의 농도가 증가함에 따라 증가하였고, CaCl₂ 용액의 농도에는 무관하였다. 고정화 효소는 효소용액을 사용했을 때보다 약 3배정도의 높은 역가를 나타내었다. 또한 β-Glucosidase를 고정화하여 첨가했을 때 원래의 가수분해 반응보다 환원당 및 포도당의 생산성을 3배 이상 증가시켰다.

The immobilization of cellulase was studied to stabilize the enzyme and thus increase the efficien-cy of the hydrolysis of cellulose. The cellulase induced from the fungus Trichoderma reesei QM 9414, was immobilized on Ca-alginate bead. The experiments were performed in order to find the optimum conditions for immobilization and understand the properties of imobilized enzyme systems. Experimenatal results show-ed that optimal concentrations of Na-alginate and CaCl₂ providing the highest activity of immobilized enzyme systems, were 3%(w/v) and 0.4m, respectively. The gel strength of the immobilized bead was increased in proportion to the concentration of Na-alginate but independent of the concentration of CaCl₂. The activity of the immobilized enzyme system was enhanced three folds compared to free enzyme system. Also, the productivities of reducing sugar and glucose were increased three folds by the addition of immobilized β-glucosidase in comparison with free β-glucosidase system.

[References]

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[1] Sternberg D, Dorval S, Biotechnol. Bioeng., 21, 181 (1979)

[2] Mandels M, Medeiros JE, Andreotti RE, Bissett FH, Biotechnol. Bioeng., 23, 2009 (1981)

[3] Ladisch MR, Lin KW, Volch M, Tsao GT, Enzyme Microb. Technol., 5, 82 (1983)

[4] Ladisch MR, Tsao GT, Enzyme Microb. Technol., 8, 66 (1986)

[5] Parisi F, Adv. Biochem. Eng., 38, 53 (1989)

[6] Mandels M, Biotechnol. Bioeng., 5, 81 (1975)

[7] Montecourt BS, Eveleigh D, Proc. 2nd Bioconv. Symp., Troy, N.Y., p. 613 (1978)

[8] Mandels M, Reese ET, J. Bacteriol., 79, 816 (1959)

[9] Messing RA, "Comprehensive Biotechnology," Vol. 2, Ch. 12, p. 203, M. Moo-Young (Ed.), Pergamon Press Ltd. (1985)

[10] Pifferi PG, Tramontini M, Malacarne A, Biotechnol. Bioeng., 33, 1258 (1989)

[11] Fujikawa S, Yokota T, Koga K, Appl. Microbiol. Biotechnol., 28, 440 (1988)

[12] Roy SK, Raha SK, Dey SK, Chakrabarty SL, Enzyme Microb. Technol., 11, 431 (1989)

[13] Jeong EJ, Lee SH, Lee YH, Korean J. Biotechnol. Bioeng., 5, 141 (1990)

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

[15] Bergmeyer HU, "Method of Enzymatic Analysis," Vol. 1, p. 457, 2nd Ed., Academic Press, NY (1974)

[16] Lowry PH, Rosebrough NJ, Farr AL, Randall RJ, J. Biol. Chem., 193, 265 (1951)

[17] Hicks GP, Updike SJ, Anal. Chem., 38, 726 (1966)

[18] Chrambach A, Rodbard D, Science, 172, 440 (1971)

[19] Miron T, Degani Y, Biochim. Biophys. Acta, 212, 362 (1970)

[20] Tosa T, Sato T, Mori T, Chibata I, Biotechnol. Bioeng., 21, 1697 (1979)