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The Effect of Organic Solvents on the Structural Stability and Catalytic Activity of Pepsin

Document Type : Article

Authors

Abstract

Pepsin enzyme is synthesized in the gastric mucosa of vertebrates. Pepsin is composed of 327 amino acid residues, with a molecular mass of 34KD. The structure is predominantly β-strand and random coil with limited α-helix. The two domains are connected via a six-stranded β-sheet plate. In this study used UV-Vis spectrophotometer pharmacia-4000 with electronic temperature controller system. Thermal stability of porcine pepsin has been investigated in the presence of organic solvents include butanol, ethanol, 1,4-Butanediol and glycerol in different concentrations (0-50% V/V) at pH=2. Tm of pepsin was decreased the presence of butanol, ethanol, 1,4-Butanediol respectively. and Tm of pepsin increased in the presence of glycerol. Also the effects of these organic solvents on the activity of porcine pepsin were studied Activity of pepsin decreased in aqueous butanol, ethanol and 1,4-Butanediol with increasing organic solvent concentration respectively and activity of pepsin increased in presence of glycerol with increasing organic solvent concentration. The changes caused in the catalytic activity by the water-miscible organic solvents include butanol, ethanol and 1,4-Butanediol may be related to structural changes, which were followed by means of thermal stability changes and also change in active site of pepsin. Glycerol also stabilized the structure of pepsin.

Keywords

References
Arakawa, T.; Timasheff, SN.; (1982) Stabilization of protein structure by sugars. Biochemistry, 21(25): 6536-6544.
Castillo, B.; Pacheco, Y.; AL-Azzam, W.; Griebenow, K.; Devi, M.; Ferrer, A.; Barletta, G.; (2005) On the activity loss of hydrolases in organic solvents: I. Rapid loss of activity of a variety of enzymes and formulations in a range of organic solvents. Journal of Molecular Catalysis B: Enzymatic, 35 (4-6): 147-153.
Chakraborty, T.; Chakraborty, I.; Moulik, SP.; Ghosh, S.; (2007) Physicochemical studies on pepsin-CTAB interaction: Energetics and structural changes. the Journal of Physical Chemistry B, 111(10): 2736-2746.
Cooper, JB.; Khan, G.; Taylor, G.; Tickle, IJ.; Blundell, TL.; (1990) X-ray analyses of aspartic proteinases: II. Three-dimensional structure of the hexagonal crystal form of porcine pepsin at 2. 3 A˚ resolution. Journal of Molecular Biology, 214 (1): 199-222.
Dunn, BM.; (2001) Overview of Pepsin‐like Aspartic Peptidases. Current Protocols in Protein Science, 21-3.
Dunn, BM.; (2002) Structure and mechanism of the pepsin-like family of aspartic peptidases. Chemical Reviews, 102 (12) : 4431-4458.
Fitzpatrick, PA.; Ringe, D.; Klibanov, AM.; (1994) X-ray crystal structure of cross-linked subtilisin carlsberg in water vs acetonitrile. Biochemical and Biophysical Research Communications, 198 (2) : 675-681.
Gupta, MN.; (1993) Enzyme function in organic solvents. EJB Reviews. Springer, 203: 25-32
Griebenow, K.; Klibanov, AM.; (1995) Lyophilization-induced reversible changes in the secondary structure of proteins. Proceedings of the National Academy of Sciences, 92(24): 10969-10976.
Kaushik, JK.; Bhat, R.; (1998) Thermal stability of proteins in aqueous polyol solutions: role of the surface tension of water in the stabilizing effect of polyols. the Journal of Physical Chemistry B, 102 (36): 7058-7066.
Mozhaev, VV.; Khmelnitsky, YL.; Sergeeva, MV.; Belova, AB.; Klyachko, NL.; Levashov, AV.; Martinek, K.; (1989) Catalytic activity and denaturation of enzymes in water/organic cosolvent mixtures. European Journal of Biochemistry, 184 (3): 597-602.
Okoniewska, M.; Tanaka, T.; Yada, RY.; (1999) The role of the flap residue, threonine 77, in the activation and catalytic activity of pepsin A. Protein Engineering, 12 (1): 55-61.
Pittz, EP.; Timasheff, SN.; (1978) Interaction of ribonuclease A with aqueous 2-methyl-2, 4-pentanediol at pH 5. 8. Biochemistry, 17(4): 615-623.
Prasad, B.; Suguna, K.; (2002) Role of water molecules in the structure and function of aspartic proteinases. Acta Crystallographica Section D: Biological Crystallography, 58: 250-259.
Sielecki, AR.; Fujinaga, M.; Read, RJ.; James, MNG.; (1991) Refined structure of porcine pepsinogen at 1·8 Å resolution. Journal of Molecular Biology, 219 (4): 671-692.
Simon, LM.; Kotormán, M.; Szabó, A.; Nemcsók, J.; Laczkó, I.; (2007) The effects of organic solvent/water mixtures on the structure and catalytic activity of porcine pepsin. Process Biochemistry, 42 (5): 909-912.
Simon, LM.; László, K.; Vertesi, A.; Bagi, K.; Szajáni, B.; (1998) Stability of hydrolytic enzymes in water-organic solvent systems. Journal of Molecular Catalysis B: Enzymatic, 4(1-2): 41-45.
Tang, J.; Sepulveda, P.; Marciniszyn, J.; Chen, KCS.; Huang, WY.; Tao, N.; Liu, D.; Lanier, JP.; (1973) Amino-acid sequence of porcine pepsin. Proceedings of the National Academy of Sciences, 70 (12): 3437-3439.
Timasheff, SN.; (1998) Control of protein stability and reactions by weakly interacting cosolvents: the simplicity of the complicated. Advances in Protein Chemistry, 51: 355-432.
Wikipedia. URL: www. wikipedia. org (accessed: 1392.8.8).
Experimental animal Biology
Volume 3, Issue 1 - Serial Number 1
January 2015
Pages 7-16
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