Chitosanases EAG1 is a classical glycoside hydrolase from Bacillus ehimensis. The previous researches showed that this Chitosanases can not only hydrolyze the b1,4-glycosidic bonds of chitosan to COS in different sizes but also keep a high catalytic activity in organic, which was useful for producing chitooligosaccharides and GlcN for use in the food and pharmacological industries. While it is instable in the liquid state. This shortcoming seriously restricts its industrial application. Here we used the modeled structure of EAG1 and the molecular modeling software package to screen the free chemical database ZINC. Moreover, the strategies including “initial filter” and consensus scoring were applied to accelerate the process and improve the success rate of virtual screening. Finally, five compounds were screened and they were purchased or synthetized to test their binding affinity against EAG1. The test results showed that one of them could inhibit the enzyme with an apparent Ki of 1.5 μM. The result may take the foundation for further inhibitor screening and design against EAG1 and the screened compound may also help to improve the liquid stability of EAG1 and expand its industrial application.
References
[1]
Vincent, L., Hemalatha, G.R., Elodie, D., Coutinho, P.M. and Bernard, H. (2014) The Carbohydrate-Active Enzymes Database (CAZy) in 2013. Nucleic Acids Research, 42, D490-D495. https://doi.org/10.1093/nar/gkt1178
[2]
Joseph, S.R. (1996) Chitosanases—Properties and Applications: A Review. Bioresource Technology, 55, 35-45.
[3]
Svendsen, A. and Clausen, I.G. (1997) Detergent Compositions Containing Protease and Novel Inhibitors for Use There.US Patent 5674833.
[4]
Schulz, P., Schwadtke, K. and Smulders, E. (1990) Process for the Preparation of a Storage-Stable Liquid Detergent Composition. US Patents US4929380A.
[5]
Stoner, M.R., Dale, D.A., Gualfetti, P.J., Becker, T., Manning, M.C., Carpenter, J.F., et al. (2004) Protease Autolysis in Heavy-Duty Liquid Detergent Formulations: Effects of Thermodynamic Stabilizers and Protease Inhibitors. Enzyme and Microbial Technology, 34, 114-125. https://doi.org/10.1016/j.enzmictec.2003.09.008
[6]
Ji, X., Zheng, Y., Wang, W., Sheng, J., Hao, J. and Sun, M. (2013) Virtual Screening of Novel Reversible Inhibitors for Marine Alkaline Protease MP. Journal of Molecular Graphics and Modelling, 46, 125-131. https://doi.org/10.1016/j.jmgm.2013.10.004
[7]
Sheng, J., Ji, X., Zheng, Y., Wang, Z. and Sun, M. (2016) Improvement in the Thermostability of Chitosanase from Bacillus ehimensis by Introducing Artificial Disulfide Bonds. Biotechnology Letters, 38, 1809-1815. https://doi.org/10.1007/s10529-016-2168-2
[8]
Dixon, M. (1953) The Determination of Enzyme Inhibitor Constants. Biochemical Journal, 55, 170-171. https://doi.org/10.1021/ci049714+
[9]
Irwin, J.J. and Shoichet, B.K. (2005) ZINC—A Free Database of Commercially Available Compounds for Virtual Screening. Journal of Chemical Information and Modeling, 45, 177-182. https://doi.org/10.1021/ci049714+
[10]
Irwin, J.J., Sterling, T., Mysinger, M.M., Bolstad, E.S. and Coleman, R.G. (2012) ZINC: A Free Tool to Discover Chemistry for Biology. Journal of Chemical Information and Modeling, 52, 1757-1768. https://doi.org/10.1021/ci3001277
[11]
Hu, X. and Shelver, W.H. (2003) Docking Studies of Matrix Metalloproteinase Inhibitors: Zinc Parameter Optimization to Improve the Binding Free Energy Prediction. Journal of Molecular Graphics and Modelling, 22, 115-126. https://doi.org/10.1016/S1093-3263(03)00153-0
[12]
Hu, X., Balaz, S. and Shelver, W.H. (2004) A Practical Approach to Docking of Zinc Metalloproteinase Inhibitors. Journal of Molecular Graphics and Modelling, 22, 293-307. https://doi.org/10.1016/j.jmgm.2003.11.002
[13]
Morris, G.M., Huey, R., Lindstrom, W., Sanner, M.F., Belew, R.K., Goodsell, D.S., et al. (2009) AutoDock4 and AutoDockTools4: Automated Docking with Selective Receptor Flexibility. Journal of Computational Chemistry, 30, 2785-2791. https://doi.org/10.1002/jcc.21256
[14]
Word, J.M., Lovell, S.C., Richardson, J.S. and Richardson, D.C. (1999) Asparagine and Glutamine: Using Hydrogen Atom Contacts in the Choice of Side-Chain Amide Orientation. Journal of Molecular Biology, 285, 1735-1747. https://doi.org/10.1006/jmbi.1998.2401
[15]
Pence, H.E. and Williams, A. (2010) ChemSpider: an Online Chemical Information Resource. Journal of Chemical Education, 87, 1123-1124. https://doi.org/10.1021/ed100697w
[16]
Wang, R., Lu, Y. and Wang, S. (2003) Comparative Evaluation of 11 Scoring Functions for Molecular Docking. Journal of Medicinal Chemistry, 46, 2287-2303. https://doi.org/10.1021/jm0203783
[17]
Lineweaver, H. and Burk, D. (1934) The Determination of Enzyme Dissociation Constants. Journal of the American Chemical Society, 56, 658-666. https://doi.org/10.1021/ja01318a036
[18]
Massaoud, M.K., Marokházi, J. and Venekei, I. (2011) Enzymatic Characterization of a Serralysin-Like Metalloprotease from the Entomopathogen Bacterium, Xenorhabdus. Biochimica et Biophysica Acta (BBA)—Proteins and Proteomics, 1814, 1333-1339. https://doi.org/10.1016/j.bbapap.2011.05.008
[19]
Si, Y., Wang, Z., Park, D., Chung, H.Y., Wang, S., Yan, L., et al. (2012) Effect of Hespaeretin on Tyrosinase: Inhibition Kinetics Integrated Computational Simulation Study. International Journal of Biological Macromolecules, 50, 257-262. https://doi.org/10.1016/j.ijbiomac.2011.11.001
