Abstract
Recently, the number of studies dealing with nonaqueous enzymology has increased. Proteases are enzymes that catalyze the hydrolysis of peptide bonds in other proteins. The 279-residue serine protease proteinase K (EC 3.4.21.14) from the fungus Tritirachium album limber belongs to the subtilisin family of enzymes. The enzyme is extensively applied for protein-related studies due to its stability at high temperature and ability to remain active in a wide range of pH (3.0–7.0). The effect of organic solvents (1,4-butanediol and butanol) on the activity of native proteinase K was studied by ultraviolet–visible (UV–Vis) spectroscopy and molecular docking. The results of spectroscopy show that 1,4-butanediol and butanol lead to increase in Vmax and decrease in Km values at low concentrations. Furthermore, they indicated that butanol is a slightly better enzyme activator in comparison with 1,4-butanediol. Molecular docking data illustrate interaction between organic solvents and PK, where hydrogen bonds and hydrophobic interactions play a major role in the PK-organic solvents complex formation. Therefore, we could conclude that these interactions lead to change in the native conformation of PK and increased enzyme activity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Babu KR, Douglas DJ (2000) Methanol-induced conformations of myoglobin at pH 4.0. Biochemistry 39:14702–14710
Betzel C, Pal GP, Saenger W (2005) Threedimensional structure of proteinase K at 0.15 nm resolution. Eur J Biochem 178(1):155–171
Bychkova VE, Dujsekina AE, Klenin SI, Tiktopulo EI, Uversky VN, Ptitsyn OB (1996) Molten globule-like state of cytochrome c under conditions simulating those near the membrane surface. Biochemistry 35:6058–6063
Ebeling W, Hennrich N, Klockow M, Metz H, Orth HD, Lang H (2005) Proteinase K from Tritirachium album limber. Eur J Biochem 47(1):91–97
FernNdez M, Fragoso A, Cao R, BaOs M, Ansorge-Schumacher M, Hartmeier W, Villalonga R (2004) Functional properties and application in peptide synthesis of trypsin modified with cyclodextrin-containing dicarboxylic acids. J Mol Catal B Enzym 31:47–52
Jany KD, Lederer G, Mayer B (1986) Amino acid sequence of proteinase K from the mold Tritirachium album Limber: proteinase K a subtilisin-related enzyme with disulfide bonds. FEBS Lett 199(2):139–144
Kamatari YO, Konno T, Kataoka M, Akasaka K (1998) The methanol-induced transition and the expanded helical conformation in hen lysozyme. Protein Sci 7:681–688
Kermasha S, Bao H, Bisakowski B (2001) Biocatalysis of tyrosinase using catechin as substrate in selected organic solvent media. J Mol Catal B Enzym 11:929–938
Klibanov AM (2001) Improving enzymes by using them in organic solvents. Nature 409(6817):241–246
Koops BC, Verheij HM, Slotboom AJ, Egmond MR (1999) Effect of chemical modification on the activity of lipases in organic solvents. Enzym Microb Technol 25:622–631
Kumar A, Attri P, Venkatesu P (2012) Effect of polyols on the native structure of α-chymotrypsin: a comparable study. Thermochim Acta 536:55–62
Laxmi D, Priyadarshy S (2002) Biotech software and internet report: the computer software. J Sci 3:5–9
Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell DS, Olson AJ (2009) J Comput Chem 30:2785–2791
Müller A, Hinrichs W, Wolf WM, Saenger W (1994) Crystal structure of calcium-free proteinase K at 1.5-A resolution. J Biol Chem 269:23108–23111
Okazaki S, Goto M, Furusaki S (2000) Surfactant–protease complex as a novel biocatalyst for peptide synthesis in hydrophilic organic solvents. Enzym Microbiol Technol 26:159–164
Palmer T (2001) Enzymes: biochemistry, biotechnology and clinical chemistry. Horwood Publishing Limited, London, pp 98–134
Powers JC, Asgian JL, Ekici ZD, James E (2002) Irreversible Inhibitors of serine, cysteine, and threonine proteases. Chem Rev 102:4639–4750
Raducan A, Cantemir AR, Puiu M, Oancea D (2012) Kinetics of hydrogen peroxide decomposition by catalase: hydroxylic solvent effects. Bioprocess Biosyst Eng 35:1523–1530
Segel IH (1993) Enzyme kinetics: behavior and analysis of rapid equilibrium and steady-state enzyme systems. Wily-Interscience, New York
Sola-penna M, Meyer-Fernandes JR (1998) Stabilization against thermal inactivation promoted by sugars on enzyme structure and function: why is trehalose more effective than other sugars? Arch Biochem Biophys 360(1):10–14
Thurmond RL, Edwards JP (2006) Drug Discov. Ser 5, 113 (Handbook of assay development in drug discovery)
Timasheff SN (1993) The control of protein stability and association by weak interactions with water: how do solvents affect these processes? Annu Rev Biophys Biomol Struct 22:67–97
Wolf WM, Bajorath J, Müller A, Raghunathan S, Singh TP, Hinrichs W, Saenger W (1991) Inhibition of proteinase K by methoxysuccinyl-Ala-Ala-Pro-Ala-chloromethyl ketone. An x-ray study at 2.2-A resolution. J Biol Chem 266:17695–17699
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yadollahi, E., Shareghi, B. & Eslami farsani, R. Molecular Aspects of the Interaction of Organic Solvents and Proteinase K: Kinetics and Docking Studies. Iran J Sci Technol Trans Sci 43, 57–62 (2019). https://doi.org/10.1007/s40995-018-0536-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s40995-018-0536-6