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تثبیت کووالانسی آنزیم پراکسیداز نوترکیب لپیدیوم درابا (LDP) بر روی ساختار آلی-فلزی روی با استفاده از گلوتارآلدئید | ||
زیست شناسی کاربردی | ||
مقاله 8، دوره 35، شماره 2 - شماره پیاپی 72، شهریور 1401، صفحه 108-125 اصل مقاله (817.84 K) | ||
نوع مقاله: مقاله پژوهشی | ||
شناسه دیجیتال (DOI): 10.22051/jab.2022.38355.1451 | ||
نویسندگان | ||
سودابه فرهادی1؛ علی ریاحی مدوار* 2؛ مجتبی مرتضوی3؛ قاسم سرگزی4 | ||
1دانشآموخته کارشناسی ارشد، گروه بیوتکنولوژی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان، ایران. | ||
2دانشیار، گروه زیست شناسی سلولی-مولکولی، دانشکده علوم پایه، دانشگاه کوثر بجنورد، بجنورد، ایران.) | ||
3استادیار، گروه بیوتکنولوژی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان، ایران. | ||
4استادیار، مرکز تحقیقات بیماریهای غیر واگیردار، دانشگاه علوم پزشکی و خدمات بهداشتی و درمانی بم، بم، ایران | ||
چکیده | ||
مقدمه: پراکسیداز لپیدیوم درابا (LDP) آنزیمی از کلاس III پراکسیدازهای گیاهی میباشد که توالی اسیدآمینهای آن با پراکسیداز ترب کوهی (HRP) بیش از 90 درصد تشابه دارد. روشها: در این پژوهش، بهمنظور اتصال محکم LDP به چارچوب آلی-فلزی روی (Zn-MOF)، بعد از بیان و تخلیص آنزیم، شرایط تثبیت با استفاده از لینکر گلوتارآلدئید بهینهسازی شد و سپس ویژگیهای فیزیکوشیمیایی، سینتیکی و پایداری آن با آنزیم آزاد مقایسه شدند. نتایج و بحث: بهترین شرایط تثبیت با بازده 67%، در غلظتهای؛ Zn-MOF 02/0 گرم، آنزیم mg/ml 75/0 و گلوتارآلدئید 2/1 دسیمولار به مدت سه ساعت انکوبهشدن بدست آمد. نتایج نشان داد که فعالیت ویژه آنزیم تثبیت شده بیش از دو برابر آنزیم آزاد افزایش یافته است و Km آن به سوبسترای TMB نسبت به آنزیم آزاد، 49 درصد کاهش یافته است. همچنین پایداری سینتیکی آنزیم تثبیت شده در برابر pH و دما نسبت به آنزیم آزاد کاهش یافت | ||
کلیدواژهها | ||
بهینهسازی؛ پایداری؛ سینتیکی؛ فعالیت ویژه | ||
عنوان مقاله [English] | ||
Covalent Immobilization of recombinant Lepidium draba peroxidase on zinc metal-organic framework using glutaraldehyde | ||
نویسندگان [English] | ||
Soudabeh Farhadi1؛ Ali Riahi-Madvar2؛ Mojtaba Mortazavi3؛ Ghasem Sargazi4 | ||
1Master's student, Department of Biotechnology, Research Institute of Advanced Science and Technology and Environmental Sciences, Post Graduate University of Industrial and Advanced Technology, Kerman, Iran. | ||
2Associate Professor, Department of Cell-Molecular Biology, Faculty of Basic Sciences, Kausar University, Bojnord, Iran.) | ||
3Assistant Professor, Department of Biotechnology, Research Institute of Advanced Science and Technology and Environmental Sciences, Postgraduate University of Industrial and Advanced Technology, Kerman, Iran | ||
4Assistant Professor, Non-Communicable Diseases Research Center, Bam University of Medical Sciences and Health Services, Bam, Iran | ||
چکیده [English] | ||
Introduction: Lepidium draba peroxidase (LDP) belonging to the class III plant peroxidases that its amino acid sequence shows over 90% similarity with horseradish peroxidase (HRP). Methods: In this study, after expression and purification of LDP, its immobilization conditions were optimized on the Zinc metal-organic framework (Zn-MOF) using glutaraldehyde as a cross-linking agent for firm binding. Then physicochemical properties, kinetic parameters and stability of the immobilized enzyme were compared with the free one. Results and discussion: The best conditions for enzyme immobilization with 67% yield were optimized at concentration of 0.02 g of Zn-MOF, 0.75mg/ml and 1.2 dM of glutaraldehyde, after 3h incubation. The results showed that the specific activity of the immobilized enzyme increased more than doubled that of free enzyme and its Km was reduced by 49% compared to the free one for TMB substrate. Also its kinetic stability reduced against pH and temperature in compared to the free enzyme. | ||
کلیدواژهها [English] | ||
Kinetics, Optimization, Specific Activity, Stability | ||
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مراجع | ||
Ai, J., Zhang, W., Liao, G., Xia, H. and Wang, D. (2016). Immobilization of horseradish peroxidase enzymes on hydrous-titanium and application for phenol removal. RSC advances 6(44): 38117-38123. Alemzadeh, I. and Nejati, S. (2009). Phenols removal by immobilized horseradish peroxidase. Journal of Hazardous Materials 166(2-3): 1082-1086. Ali, S. M. U., Nur, O., Willander, M. and Danielsson, B. (2010). A fast and sensitive potentiometric glucose microsensor based on glucose oxidase coated ZnO nanowires grown on a thin silver wire. Sensors and Actuators B: Chemical 145(2): 869-874. Altikatoglu Yapaoz, M. and Attar, A. (2020). An accomplished procedure of horseradish peroxidase immobilization for removal of acid yellow 11 in aqueous solutions. Water Science and Technology 81(12): 2664-2673. Ansari, S. A. and Husain, Q. (2012). Potential applications of enzymes immobilized on/in nano materials: A review. Biotechnology advances 30(3): 512-523. Barragán, L. P., Buenrostro-Figueroa, J., González, C. A. and Marañon, I. (2016). Production, Stabilization, and Uses of Enzymes From Fruit and Vegetable Byproducts. Biotransformation of Agricultural Waste and By-Products, Elsevier: 271-286. Bayramoğlu, G. and Arıca, M. Y. (2008). Enzymatic removal of phenol and p-chlorophenol in enzyme reactor: horseradish peroxidase immobilized on magnetic beads. Journal of hazardous materials 156(1-3): 148-155. Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry 72(1-2): 248-254. Bilal, M., Adeel, M., Rasheed, T., L, H.M.N. (2019) Multifunctional metal–organic frameworks-based biocatalytic platforms: recent developments and future prospects. Journal of Materials Research and Technology 8(2): 2359-2371. Cano, À., Minguillón, C. and Palet, C. (2006). Immobilization of endo-1, 4-β-xylanase on polysulfone acrylate membranes: Synthesis and characterization. Journal of membrane science 383:(2-1)280. Cao, S.-L., Yue, D.-M., Li, X.-H., Smith, T. J., Li, N., Zong, M.-H., Wu, H., Ma, Y.-Z. and Lou, W.-Y. (2016a). Novel nano-/micro-biocatalyst: soybean epoxide hydrolase immobilized on UiO-66-NH2 MOF for efficient biosynthesis of enantiopure (R)-1, 2-octanediol in deep eutectic solvents. ACS Sustainable Chemistry & Engineering 4(6): 3586-3595. Cao, S., Xu, P., Ma, Y., Yao, X., Yao, Y., Zong, M., Li, X. and Lou, W. (2016). Recent advances in immobilized enzymes on nanocarriers. Chinese Journal of Catalysis 37(11): 1814-1823. Chae, H. J., In, M.-J. and Kim, E. Y. (1998). Optimization of protease immobilization by covalent binding using glutaraldehyde. Applied biochemistry and biotechnology 73(2-3): 195-204. Chakraborty, S., Rusli, H., Nath, A., Sikder, J., Bhattacharjee, C., Curcio, S. and Drioli, E. (2016). Immobilized biocatalytic process development and potential application in membrane separation: a review. Critical reviews in biotechnology 36(1): 43-58. Chang, T. (1971). Stabilization of enzyme by microencapsulation with a concentrated protein solution or by crosslinking with glutaraldehyde. Biochem Biophys Res Com 44(6): 1531-1533. Cipolatti, E. P., Valerio, A., Henriques, R. O., Moritz, D. E., Ninow, J. L., Freire, D. M., Manoel, E. A., Fernandez-Lafuente, R. and de Oliveira, D. (2016). Nanomaterials for biocatalyst immobilization–state of the art and future trends. RSC Advances 6(106): 104675-104692. Danson, M. J. and Hough, D. W. (1998). Structure, function and stability of enzymes from the Archaea. Trends in microbiology 6(8): 307-314. Dhruvaraj, M. (2017). Role of peroxidase in clinical assays: A short review. Journal of Clinical Nutrition 3(2): 14. Dong, L., Ge, C., Qin, P., Chen, Y. and Xu, Q. (2014). Immobilization and catalytic properties of candida lipolytic lipase on surface of organic intercalated and modified MgAl-LDHs. Solid State Sciences 31: 8-15. Farhadi, S., Riahi-Madvar, A., Sargazi, G. and Mortazavi, M. (2021). Immobilization of Lepidium draba peroxidase on a novel Zn-MOF nanostructure. International Journal of Biological Macromolecules 173: 366-378. Fattahian, Y., Riahi-Madvar, A., Mirzaee, R., Asadikaram, G. and Rahbar, M. R. (2017). In silico locating the immune-reactive segments of Lepidium draba peroxidase and designing a less immune-reactive enzyme derivative. Computational biology and chemistry 70: 21-30. Fattahian, Y., Riahi-Madvar, A., Mirzaee, R., Torkzadeh-Mahani, M. and Asadikaram, G. (2017). Heterologous expression, purification and characterization of a peroxidase isolated from Lepidium draba. The protein journal 36(6): 461-471. Fernandez-Lafuente, R., Rosell, C., Rodriguez, V. and Guisan, J. (1995). Strategies for enzyme stabilization by intramolecular crosslinking with bifunctional reagents. Enzyme and microbial technology 17(6): 517-523. Garcia‐Galan, C., Berenguer‐Murcia, Á., Fernandez‐Lafuente, R. and Rodrigues, R. C. (2011). Potential of different enzyme immobilization strategies to improve enzyme performance. Advanced Synthesis & Catalysis 3532904-2885:(16). Getman, R. B., Bae, Y.-S., Wilmer, C. E. and Snurr, R. Q. (2012). Review and analysis of molecular simulations of methane, hydrogen, and acetylene storage in metal–organic frameworks. Chemical reviews 112(2): 703-723. Hanefeld, U., Gardossi, L. and Magner, E. (2009). Understanding enzyme immobilisation. Chemical Society Reviews 38(2): 453-468. Hassanzadeh, J., Khataee, A. and Eskandari, H. (2018). Encapsulated cholesterol oxidase in metal-organic framework and biomimetic Ag nanocluster decorated MoS2 nanosheets for sensitive detection of cholesterol. Sensors and Actuators B: Chemical 259: 402-410. Hernandez, K. and Fernandez-Lafuente, R. (2011). Control of protein immobilization: Coupling immobilization and site-directed mutagenesis to improve biocatalyst or biosensor performance. Enzyme and microbial technology 48(2): 107-122. Hirsh, S., Bilek, M., Nosworthy, N., Kondyurin, A., Dos Remedios, C. and McKenzie, D. (2010). A comparison of covalent immobilization and physical adsorption of a cellulase enzyme mixture. Langmuir 26(17): 14380-14388. Kinsley, C. and Nicell, J. A. (2000). Treatment of aqueous phenol with soybean peroxidase in the presence of polyethylene glycol. Bioresource Technology 73(2): 139-146. Klibanov, A. M. (2001). Improving enzymes by using them in organic solvents. nature 409(6817): 241. Krainer, F. W., Pletzenauer, R., Rossetti, L., Herwig, C., Glieder, A. and Spadiut, O. (2014). Purification and basic biochemical characterization of 19 recombinant plant peroxidase isoenzymes produced in Pichia pastoris. Protein expression and purification 95: 104-112. Li, J., Chen, X., Xu, D. and Pan, K. (2019). Immobilization of horseradish peroxidase on electrospun magnetic nanofibers for phenol removal. Ecotoxicology and environmental safety 170: 716-721. Magnan, E., Catarino, I., Paolucci-Jeanjean, D., Preziosi-Belloy, L. and Belleville, M.-P. (2004). Immobilization of lipase on a ceramic membrane: activity and stability. Journal of Membrane Science 241(1): 161-166. Mateo, C., Palomo, J. M., Fernandez-Lorente, G., Guisan, J. M. and Fernandez-Lafuente, R. (2007). Improvement of enzyme activity, stability and selectivity via immobilization techniques. Enzyme and microbial technology 40(6): 1451-1463. Michałowicz, J. and Duda, W. (2007). Phenols--Sources and Toxicity. Polish Journal of Environmental Studies 16(3). Mohamad, N. R., Marzuki, N. H. C., Buang, N. A., Huyop, F. and Wahab, R. A. (2015). An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes. Biotechnology & Biotechnological Equipment 29(2): 205-220. Mohamed, S. A., Al-Harbi, M. H., Almulaiky, Y. Q., Ibrahim, I. H. and El-Shishtawy, R. M. (2017). Immobilization of horseradish peroxidase on Fe3O4 magnetic nanoparticles. Electronic Journal of Biotechnology 27: 84-90. Nadar, S. S. and Rathod, V. K. (2020). Immobilization of proline activated lipase within metal organic framework (MOF). International journal of biological macromolecules 152: 1108-1112. NOUROUZIAN, D. (2003) Enzyme immobilization: the state of art in biotechnology. Pantić, N., Prodanović, R., Đurđić, K. I., Polović, N., Spasojević, M. and Prodanović, O. (2021). Optimization of phenol removal with horseradish peroxidase encapsulated within tyramine-alginate micro-beads. Environmental Technology & Innovation 21: 101211. Radzicka, A. and Wolfenden, R. (1995). A proficient enzyme. Science 267(5194): 90-93. Rodrigues, R. C., Ortiz, C., Berenguer-Murcia, Á., Torres, R. and Fernández-Lafuente, R. (2013). Modifying enzyme activity and selectivity by immobilization. Chemical Society Reviews 42(15): 6290-6307. Sahin, S. and Ozmen, I. (2020). Covalent immobilization of trypsin on polyvinyl alcohol-coated magnetic nanoparticles activated with glutaraldehyde. Journal of pharmaceutical and biomedical analysis 184: 113195. Sakai-Kato, K., Kato, M., Ishihara, K. and Toyo'oka, T. (2004). An enzyme-immobilization method for integration of biofunctions on a microchip using a water-soluble amphiphilic phospholipid polymer having a reacting group. Lab on a Chip 4(1): 4-6. Sanz, V., De Marcos, S., Castillo, J. R. and Galbán, J. (2005). Application of molecular absorption properties of horseradish peroxidase for self-indicating enzymatic interactions and analytical methods. Journal of the American Chemical Society 127(3): 1038-1048. Sargazi, G., Afzali, D., Ebrahimi, A. K., Badoei-Dalfard, A., Malekabadi, S. and Karami, Z. (2018). Ultrasound assisted reverse micelle efficient synthesis of new Ta-MOF@ Fe3O4 core/shell nanostructures as a novel candidate for lipase immobilization. Materials Science and Engineering: C 93: 768-775. Sheldon, R. A. (2007). Enzyme immobilization: the quest for optimum performance. Advanced Synthesis & Catalysis 349(8‐9): 1289-1307. Silva, J., Macedo, G., Rodrigues, D., Giordano, R. and Gonçalves, L. (2012). Immobilization of Candida antarctica lipase B by covalent attachment on chitosan-based hydrogels using different support activation strategies. Biochemical Engineering Journal 60: 16-24. Stephens, I. E., Bondarenko, A. S., Bech, L. and Chorkendorff, I. (2012). Oxygen Electroreduction Activity and X‐Ray Photoelectron Spectroscopy of Platinum and Early Transition Metal Alloys. ChemCatChem 4(3): 341-349. Tatsumi, K., Wada, S. and Ichikawa, H. (1996). Removal of chlorophenols from wastewater by immobilized horseradish peroxidase. Biotechnology and Bioengineering 51(1): 126-130. Tischer, W. and Wedekind, F. (1999). Immobilized enzymes: methods and applications. Biocatalysis-from discovery to application, Springer: 95-126. Tosa, T., Mori, T., Fuse, N. and Chibata, I. (1966). Studies on continuous enzyme reactions. I. Screening of carriers for preparation of water-insoluble aminoacylase. Enzymologia 31(4): 214. Tukel, S. S. and Alptekin, O. (2004). Immobilization and kinetics of catalase onto magnesium silicate. Process Biochemistry 39(12): 2149-2155. Xia, G. H., Cao, S. L., Xu, P., Li, X. H., Zhou, J., Zong, M. H. and Lou, W. Y. (2017). Preparation of a nanobiocatalyst by efficiently immobilizing Aspergillus niger lipase onto magnetic metal–biomolecule frameworks (BioMOF). ChemCatChem 9(10): 1794-1800. Xie, Y., Chen, C., Ren, X., Wang, X., Wang, H., Wang, X. (2019) Emerging natural and tailored materials for uranium-contaminated water treatment and environmental remediation. Progress in Materials Science103: 180-234. Yujun, S., Konggang, Q., Chao, Z., Jinsong, R. and Xiaogang, Q. (2010). Graphene oxide: intrinsic peroxidase catalytic activity and its application to glucose detection. Adv. Mater. 22: 2206-2210. Zhang, Y.-W., Tiwari, M. K., Jeya, M. and Lee, J.-K. (2011). Covalent immobilization of recombinant Rhizobium etli CFN42 xylitol dehydrogenase onto modified silica nanoparticles. Applied microbiology and biotechnology 90(2): 499-507. Zhou, H. (2012). J. r. long, OM yaghi. Chem. Rev 112: 673. Zhou, H. (2014). C.; Kitagawa, S. Metal− Organic Frameworks (MOFs). Chem. Soc. Rev 43: 5415-5418. Zhou, Q. Z. and Chen, X. D. (2001). Immobilization of β-galactosidase on graphite surface by glutaraldehyde. Journal of food engineering 48(1): 69-74. Zucca, P. and Sanjust, E. (2014). Inorganic materials as supports for covalent enzyme immobilization: methods and mechanisms. Molecules 19(9): 14139-14194.
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