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UPSI Digital Repository (UDRep)
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| Abstract : Universiti Pendidikan Sultan Idris |
| Aminoacylase (EC 3.5.1.14) hydrolyzes N–acetylated amino acids to produce amino acids. Although thermostable aminoacylase has been commercially produced since 2004, there was a knowledge gap in the field of understanding aminoacylase thermostability from a structural point of view. This study investigated the physical and structural properties of the purified thermostable aminoacylase SZN. The spectropolarimetry data for structural determination has indicated a gradual decrease of α-helix from 36 to 27.6%, followed by tremendous disorientation of the structure at the transition of temperatures from 60 to 70°C (27.6 to 19.5%). In contrast, the percentage of β-sheet has increased steadily over the tested temperatures. The α-helix, where notable metal binding and catalytic residues are located, was totally weakened at temperatures above 70?C, thus resulted in loss of activity. The loss of the α-helical structure could further explain drastic deterioration of activity at temperatures beyond 70?C. The activity of aminoacylase SZN was enhanced by divalent metal ions, such as Mn2+ and Cu2+, and inhibited by detergent Triton-X-100. As a conclusion, the isolated aminoacylase SZN was characterized as a thermostable enzyme based on the α-helical structure integrity and functional stability in high temperatures. This enzyme could be used as an alternative enzyme for bioindustries in view of its activity enhancement in high temperatures and stability in various tested inhibitors |
| References |
Adenan. S., Wong, C. F., Zain, H. H. M, Azziz, S. S. S. A., & Rahman, R. N. Z. R. A. 2018. Isolation, identification and molecular characterization of thermophilic aminoacylase from Geobacillus sp. strain SZN. International Journal of Science Environment and Technology 7(5): 1483–1494. Bradford, M. M. 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principles of protein-dye binding. Analytical Biochemistry 72: 248-254. Cheng, M., Takenaka, S., Aoki, S., Murakami, S., & Aoki, K. 2009. Purification and characterization of an eggshell membrane decomposing protease from Pseudomonas aeruginosa strain ME-4. Journal of Bioscience and Bioengineering 107: 373-378. Deleage G. & Geourjon. C. 1995. SOPMA: significant improvements in protein secondary structure prediction by concensus prediction from multiple allignments. Computer Applications in the Biosciences 11(6): 681-684. Gupta, A., Roy, I., Khare, S. K., & Gupta, M. N. 2005. Purification and characterization of a solvent stable protease from Pseudomonas aeruginosa PseA. Journal of Chromatography A 1069: 155-161. Hollingsworth, E. J., Isupov, M. N., & Littlechild, J. A. 2002. Crystallization and preliminary X-ray diffraction analysis of L-aminoacylase from the hyperthermophilic archaeon Thermococcus litoralis. Acta Crystallographica Section D Biological Crystallography 58(3): 507–510. Joo, H.-S. & Choi, J. W. 2012. Purification and characterization of a novel alkaline protease from Bacillus horikoshii. Journal of Microbiology and Biotechnology 22(1): 58–68. Knape, M. J., Bertinetti, D., Ballez, M., Burghardt, N. C., Zimmermann, B., Kornev, A. P., & Herberg, F. W. 2017. Divalent metal ions control activity and inhibition of protein kinases. Metallomics 9(11): 1576-1584. Koreishi, M., Nakatani, Y., Ooi, M., Imanaka, H., Imamura, K., & Nakanishi, K. 2009. Purification, characterization, molecular cloning, and expression of a new aminoacylase from Streptomyces mobaraensis that can hydrolyze N-(Middle/Long)-chain-fatty-acyl-L-amino acids as well as N-Short-chain-acyl-L-amino acids. Bioscience, Biotechnology, and Biochemistry 73 (9): 1940–1947. Lin, L. L., Chen, M. H., Chien, H. C., Kan, S. C., Chen, C. C., Hu, H. Y., & Hsu, W. H. 2007. Characterization of a bifunctional aminoacylase/carboxypeptidase from radioresistant bacterium Deninococcus radiodurans R1. Journal of Biotechnology 128: 322-334. Lindner, H. A., Lunin, V. V, Alary, A., Hecker, R. and Cygler, M. 2003. Essential roles of zinc ligation and enzyme dimerization for catalysis in the aminoacylase-1/M20 Family. Journal of Biological Chemistry 278(45): 44496–44504. Moore, S. & Stein, W. H. 1948. Photometric ninhydrin method for use in the chromatography of amino acids. Journal of Biological Chemistry 176: 367-388. Rahman, R. N. Z. R. A., Wong, C. F, Salleh, A. B., & Basri, M. 2011. Role of α-helical structure in organic solvent-activated homodimer of elastase strain K. International Journal of Molecular Sciences 12: 5797–5814. Raussens, V., Ruysschaert, J. M., & Goormaghtigh, E. 2003. Protein concentration is not an absolute prerequisite for the determination of secondary structure from circular dichroism spectra: a new scaling method. Analytical Biochemistry 319(1): 114-21. Research & Market. 2015. Amino acids market analysis by product (l-glutamate, lysine, methionine, threonine, tryptophan, leucine, iso-leucine, valine, glutamine, arginine, glycine, phenylalanine, tyrosine, citrulline, creatine, proline, serine), by source (plant-based & animal-based), by application (animal feed, food & dietary supplements & pharmaceuticals) and segment forecasts to 2022. Amino acids market size, share, trends, global industry report, 2022. Retrieved from https://www.grandviewresearch. com/industry-analysis/amino-acids-market. Robinson, P. K. 2015. Enzymes: principles and biotechnological applications. Essays in Biochemistry 59: 1–41. Sakanyan, V., Desmarez, L., Legrain, C., Charlier, D., Mett, I., Kochikyan, A., Savchenko, A., Boyen, A., Fulmagne, P., Pierard, A., & Glansdorff, N. 1993. Gene cloning, sequence analysis, purification, and characterization of a thermostable aminoacylase from Bacillus strearothermophilus. Applied and Environmental Microbiology 59: 3878-3888. Story, S. V., Grunden, A. M., & Adams, M. W. 2001. Characterization of an aminoacylase from the hyperthermophilic archaeon Pyrococcus furiosus. Journal of Bacteriology 18: 4259-4268. Tang, Z., Yu, J., Zhuo, Q., He, B., Wang, Z. & Zhuo, H. 1995. Secondary structure circular of holo- and FT-Raman apo-aminoacylase spectroscopy from prediction of aminoacylase. Journal of Biochemistry 709: 706–709. Tanimoto, K., Higashi, N., Nishioka, M., Ishikawa, K., and Taya, M. 2008. Characterization of thermostable aminoacylase from hyperthermophilic archaeon Pyrococcus horikoshii. FEBS Journal 275: 1140–1149.
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