Similar articles

Abstract: The solubilization of elastin by Bacillus licheniformis elastase cannot be analyzed by conventional kinetic methods because the biologically relevant substrate is insoluble and the concentration of enzyme-substrate complex has no physical meaning. In this paper we report the optimization of elastolysis conditions and analysis of elastolytic kinetics. Our results indicated that the hydrolyzing temperature and time are very important factors affecting elastolysis rate. The optimized conditions using central composite design were as follows: elastolysis temperature 50 °C, elastase concentration 1×10⁴ U/ml, elastin 80 mg, elastolytic time 4 h. Investigation of the effects of substrate content, elastase concentration and pH was also revealed that low or high elastin content inhibits the elastolysis process. Increasing elastase improves elastin degradation, but high elastase may change the kinetics characterization. Alkaline environment can decrease elastin degradation rate and pH may affect elastolysis by changing elastase reaction pH. To further elucidate the elastolysis process, the logistic model was used to elastolysis kinetics study showing clearly that the logistic model can reasonably explain the elastolysis process, especially under lower elastase concentration. However, there is still need for more investigations with the aid of other methods, such as biochemical and molecular methods.

[1] Adler-Nissen, J., 1986. Enzymic Hydrolysis of Food Protein. Elsevier Applied Science Publishers, London.

[2] Baici, A., 1992. Current Topics in Rehabilitation. In: Grassi, C., Travis, J., Casali, L., Luisetti, M. (Eds.), Biochemistry of Pulmonary Emphysema. Spring-Verlag, Berlin, p.81-91.

[3] Bradford, M.M., 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72(1-2):248.

[4] Chen, Q.H., He, G.Q., Molhtar, A.M.A., 2002. Optimization of medium composition for the production of elastase by Bacillus sp. EL31410 with response surface methodology. Enzyme Microb. Technol., 30(5):667-672.

[5] Chen, Q.H., He, G.Q., Wu, Y.L., 2003. Screening of elastase-producing strains and primary studies on elastase fermentation conditions. Journal of Zhejiang University (Agriculture and Life Science), 59(1):59-62 (in Chinese).

[6] Chen, Q.H., He, G.Q., Schwarz, P., 2004. A kinetic model for elastase production by Bacillus sp. EL31410. Journal of Agricultural and Food Chemistry, 52(11):3356-3359.

[7] Du, J.Z., Shu, B.G., Wei, X.C., 1991. Enzyme Structure and Reaction Mechanism. Peking University Press, Beijing, p.104-180 (in Chinese).

[8] Hall, D.A., Czerkawaki, J.W., 1961. The reaction between elastase and elastic tissue. Biochem. J., 80:128-136.

[9] Heather, M.M., Howard, G.W., Caroline, A.O., Robert, A.S., Edward, J.C., 1999. Interaction between leukocyte elastase and elastin: quantitative and catalytic analyses. Biochimica et Biophysica Acta, 1430(2):179-190.

[10] Lahl, R., 1994. Enzymatic production of protein hydrolysates for food uses. Food Technol., 138:68-71.

[11] Lonky, S.A., Wohl, H., 1983. Regulation of elastolysis of insoluble elastin by human leukocyte elastase: stimulation by lysine-rich ligands, anionic detergents, and ionic strength. Biochemistry, 22(15):3714-3720.

[12] Ma, A.Y.M., Ooraikul, B., 1986. Optimization of enzymatic hydrolysis of canola meal with response surface methodology. J. Food Proc. Preserv., 10:99-113.

[13] Mecham, R.P., Broekelmann, T.J., Fliszar, C.J., Shapiro, S.D., Welgus, H.G., Senior, R.M., 1997. Elastin degradation by matrix metalloproteinases. J. Biol. Chem., 272(29):18071-18076.

[14] Ozaki, H., Shiio, I., 1975. Purification and properties of elastolytic enzyme from Flavobacterium immotum. J. Biochem., 77:171-180.

[15] Partridge, S.M., Davis, H.F., 1948. The chemistry of connective tissues. Biochemical J., 43:387.

[16] Sachar, L.A., 1955. Photometry method for estimation of elastase activity. Proc. Soc. Expeti. Biol. Med., 90:5323-5325.

[17] Sharon, S.R., Jennifer, L.R., Bryan, N.B., 1997. Identification of an elastolytic protease in stationary phase culture filtrates of M. tuberculosis. FEMS Microbiology, 151(1):59-64.

[18] Tsai, Y.C.H., Jung, R.Y., Lin, S.F., 1988. Production and further characterization of an alkaline elastase production by alkalophilic Bacillus strain YaB. Appl. Environm. Microbial., 54(1):3156-3161.

[19] Tsuzuki, H., Oka, T., 1965. Pseudomonas aeruginosa elastase: isolation, crystallization and preliminary characterization. J. Biol. Chem., 8:3295-3303.