화학공학소재연구정보센터
Korean Journal of Chemical Engineering, Vol.19, No.2, 261-266, March, 2002
Immobilization of GL-7-ACA Acylase for the Production of 7-ACA
E-mail:
Glutaryl-7-aminocephalosporanic acid(GL-7-ACA) acylase is an important enzyme for the production of 7-ACA (7-aminocephalosporanic acid). For an efficient immobilization of GL-7-ACA acylase, various carriers were tested. A high-porous hydrophilic carrier (FPHA) among various carriers tested was found to be the best for the immobilization of GL-7-ACA acylase. In order to develop an effective immobilization method of GL-7-ACA acylase, the parameters that affect the immobilization of GL-7-ACA acylase were also investigated under different conditions of buffer solution and different concentrations of glutaraldehyde. The highest value of GL-7-ACA acylase activity (70 Unit/g-matrix) was obtained when immobilized with 1% glutaraldehyde in a 0.1M Tris buffer (pH 8.0). Also, in order to enhance the activity of the immobilized GL-7-ACA acylase, unreacted aldehyde groups were quenched by reaction with a low molecular weight agent such as L-lysine after immobilization. The highest activity of immobilized GL-7-ACA acylase was obtained at 0.1% of L-lysine. The immobilized GL-7-ACA acylase was tested for long-term stability and it was found that the activity was retained at about 62% of the initial value after 72 times of reuse at 25 ℃.
  1. Alfani F, Cantarella M, Cifoni D, Spreti N, Germani R, Savelli G, Bioprocess Eng., 21, 13 (1999) 
  2. Alfani F, Cantarella M, Cutarella N, Gallifuoco A, Golini P, Bianchi D, Biotechnol. Lett., 19(2), 175 (1997) 
  3. Bianchi D, Golini P, Bortolo R, Battistel E, Tassinari R, Cesti P, Enz. Microb. Technol., 20, 368 (1997) 
  4. Chae HJ, In MJ, Kim EY, Appl. Biochem. Biotechnol., 73(2-3), 195 (1998)
  5. Battistel E, Bianchi D, Bortolo R, Bonoldi L, Appl. Biochem. Biotechnol., 69(1), 53 (1998)
  6. Gee KB, Choi CY, Korean J. Chem. Eng., 1(1), 13 (1984)
  7. Kwon DY, Rhee JS, Korean J. Chem. Eng., 1(2), 153 (1984)
  8. Lowry OH, Rosebrough NL, Farr AL, Randall RJ, J. Biol. Chem., 193, 265 (1951)
  9. Moly Eldin MS, Schroen CGPH, Janssen AEM, Mita DG, Tramper J, J. Mol. Catal., 10, 445 (2000) 
  10. Park SW, Kim YI, Chung KH, Kim SW, Process Biochem., 37(2), 153 (2001) 
  11. Roberto FLV, Joes MG, Enz. Microb. Technol., 23, 28 (1998) 
  12. Shibuya Y, Matsumoto K, Fujh T, Agric Biol. Chem., 45, 2225 (1981)
  13. Tsuzuki K, Komatsy K, Ichikawa S, Shibuya Y, Nippon Nogeikagaku Kaishi, 63, 1847 (1989)
  14. Tumuturk H, Aksoy S, Hasici N, Food Chem., 68, 259 (2000) 
  15. Wilhem T, Wedekind F, Top. Curr. Chem., 200, 95 (1999)