Extraction and Characterization of Urease from Durio zibethinus L
Abstract
Urease is a biocatalyst that serves to hydrolyze urea into ammonia and carbon dioxide. Since it is an imported product, the price of urea is still high. Urease can be found in grains. One of the grains that has not been explored for its urease content is durian (Durio zibethinus L.) seeds. This study aims to determine the effect of germination time on the activity of urease from durian seeds and its characteristics including the effect of pH, incubation temperature, enzymatic reaction time, addition of EDTA and metals, and storage time on the activity of urease from durian seeds. The first step of this study was seed germination which was carried out in the dark for 0, 3, 5, 7, and 9 days. Durian seed sprouts were extracted by mashing them using a mortar and pestle. They were then homogenized using a stirrer and centrifuged in a cold state. The crude urease extract obtained was then tested for its activity using the Nessler method. The acquired data was tested statistically using ANOVA. The results showed that the activity of urease from durian seeds was optimum at 3-day germination time, pH 7, incubation temperature at 30 °C, and 15-minute enzymatic reaction time with an activity of 163.6 U/mL. Urease is a metalloenzyme with its inhibitor being the Cu2+ and Na+ metal ions and its activator being Ba2+ metal ion. Tukey's test analysis showed that the effect of urease storage time at 4 °C resulted in a stable urease activity for 8 days while at room temperature it decreased its activity significantly to 72.8%.
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References
Akhtaruzzaman, M., Mozumder, N. H. M. R., Jamal, R., Rahman, A., & Rahman, T. (2012). Isolation and characterization protease enzyme from leguminous seeds. Agricultural Science Research Journals, 2(8), 434–440.
Ambarsari, H., Asriyani, L., & Ridlo, A. (2020). Isolasi dan produktivitas bakteri ureolitik dari sedimen Muara Sungai Citarum (Isolation and productivity of ureolytic bacteria from Citarum River Estuary sediments). Jurnal Teknologi Lingkungan, 21(2), 147–156.
Bahri, S., & Mirzan, M. (2012). Karakterisasi enzim amilase dari kecambah biji jagung ketan ( Zea mays ceratina L .) [Characterization of amylase enzyme from glutinous corn seed (Zea mays ceratina L.) Sprouts]. Jurnal Natural Science, 1(1), 132–143.
Bano, S., Qader, S. A. U., Aman, A., Syed, M. N., & Azhar, A. (2011). Purification and characterization of novel α-amylase from Bacillus subtilis KIBGE HAS. AAPS PharmSciTech, 12(1), 255–261. https://doi.org/10.1208/s12249-011-9586-1
BPS. (2021). Produksi tanaman buah-buahan 2020 (Fruit crop production 2020). Badan Pusat Statistik.
Bedan, D. S. (2020). Extraction, precipitation and characterization of urease from Vicia faba L. Al-Mustansiriyah Journal of Science, 31(1), 9. https://doi.org/10.23851/mjs.v31i1.555
Chouhan, S., Vishnu Priya, V., & Gayathri, R. (2018). Extraction and partial purification of urease enzyme from jack fruit. International Journal of Research in Pharmaceutical Sciences, 9(2), 438–441. https://doi.org/10.26452/ijrps.v9i2.1515
Dixon, N. E., Riddles, P. W., Gazzola, C., Blakeley, R. L., & Zerner, B. (1980). Jack bean urease (EC 3.5.1.5). V. On the mechanism of action of urease on urea, formamide, acetamide, N-methylurea, and related compounds. Canadian Journal of Biochemistry, 58(12), 1335–1344. https://doi.org/10.1139/o80-181
El-hefnawy, M. E., Sakran, M., Ismail, A. I., & Aboelfetoh, E. F. (2014). Extraction , purification , kinetic and thermodynamic properties of urease from germinating Pisum Sativum L . seeds. BMC Biochemistry, 15(15), 1–8.
Habibie, F. M., Wardani, A. K., & Nurcholis, M. (2014). Isolation and molecular identification of thermophilic microorganism producing xylanase from Hot Mud Disaster Lapindo. Jurnal Pangan Dan Agroindustri, 2(4), 231–238.
Iyer, P. ., Priya, V. V., & Gayathri, R. (2018). Assessment of urease activity in Pisum sativum seeds. Drug Invention Today, 10(9), 1810–1813.
Jeong, H., Park, J., & Kim, H. (2013). Determination of NH+ in environmental water with interfering substances using the modified Nessler method. Journal of Chemistry, 2013. https://doi.org/10.1155/2013/359217
Kaur, H., Kumar, S., & Verma, N. (2014). Enzyme-based colorimetric and potentiometric biosensor for detecting Pb (II) ions in milk. Brazilian Archives of Biology and Technology, 57(4), 613–619. https://doi.org/10.1590/S1516-8913201402160
Kiran, S., Singh, A., Prabha, C., & Kumari, S. (2018). Isolation and characterization of thermostable amylase producing bacteria from Hot Springs of Bihar, India. International Journal of Pharma Medicine and Biological Sciences, 7(2), 28–34.
Krajewska, B. (2009). Ureases I. Functional, catalytic and kinetic properties: A review. Journal of Molecular Catalysis B: Enzymatic, 59(1–3), 9–21. https://doi.org/10.1016/j.molcatb.2009.01.003
Kumari, N., Jain, V., & Malhotra, S. (2013). Purification and characterization of extracellular acidophilic-amylase from Bacillus cereus MTCC 10205 isolated from soil. African Journal of Microbiology Research, 7(48), 5440–5448. https://doi.org/10.5897/ajmr12.1371
Liu, J., Xu, Y., Nie, Y., & Zhao, G. A. (2012). Optimization production of acid urease by Enterobacter sp. in an approach to reduce urea in Chinese rice wine. Bioprocess and Biosystems Engineering, 35(4), 651–657. https://doi.org/10.1007/s00449-011-0643-7
Modolo, L. V, Souza, A. X. De, Horta, P., Araujo, D., & Fa´tima, A. de. (2015). An overview on the potential of natural products as ureases inhibitors : A review q. Journal of Advanced Research, 6(1), 35–44. https://doi.org/10.1016/j.jare.2014.09.001
Nurfiana, F., Mukaromah, U., Jeannisa, V. C., & Putra, S. (2009). Pembuatan bioethanol dari biji durian sebagai sumber energi alternatif (The making of bioethanol from durian seeds as an alternative source of energy). Seminar Nasional V SDM Teknologi Nuklir, November, 669–676.
Patri, M. Y. (2018). Penentuan kadar ammonia (NH3) pada limbah cair K-36 dalam rangka pengendalian pencemaran lingkungan (Determination of ammonia (NH¬¬3) level in K-36 liquid waste for controlling environmental pollution). ALKIMIA : Jurnal Ilmu Kimia Dan Terapan, 2(2), 32–36. https://doi.org/10.19109/alkimia.v2i2.2998
Pervin, M. S., Jahan, M. G. S., Rana, A. Y. K. M., Sana, N. K., Rahman, M. H., & Shaha, R. K. (2013). Effects of some environmental variables on urease in germinating chickpea ( Cicer arietinum L .) Seed. Journal of Stress Physiology & Biochemistry, 9(3), 345–356.
Postal, M., Martinelli, A. H. S., Becker-Ritt, A. B., Ligabue-Braun, R., Demartini, D. R., Ribeiro, S. F. F., Pasquali, G., Gomes, V. M., & Carlini, C. R. (2012). Antifungal properties of Canavalia ensiformis urease and derived peptides. Peptides, 38(1), 22–32. https://doi.org/10.1016/j.peptides.2012.08.010
Saboury, A. A. (2010). A Thermodynamic Study of the interaction between urease and copper ions. Journal of Sciences, Islamic Republic of Iran, 21(1), 15–20. https://doi.org/10.22059/jsciences.2010.20134
Singh, A. K., Singh, M., & Verma, N. (2017). Extraction, purification, kinetic characterization and immobilization of urease from Bacillus sphaericus MTCC 5100. Biocatalysis and Agricultural Biotechnology, 12, 341–347. https://doi.org/10.1016/j.bcab.2017.10.020
Sutopo, L. (2004). Teknologi Benih (Seed Technology). Rajawali.
Tallapragada, P., Dikshit, R., Jadhav, A., & Sarah, U. (2017). Partial purification and characterization of amylase enzyme under solid state fermentation from Monascus sanguineus. Journal of Genetic Engineering and Biotechnology, 15(1), 95–101. https://doi.org/10.1016/j.jgeb.2017.02.003
Wu, X., Wang, Y., Tong, B., Chen, X., & Chen, J. (2018). Purification and biochemical characterization of a thermostable and acid-stable alpha-amylase from Bacillus licheniformis B4-423. International Journal of Biological Macromolecules, 109, 329–337. https://doi.org/10.1016/j.ijbiomac.2017.12.004
Zusfahair, Ningsih, D. R., Fatoni, A., & Pertiwi, D. S. (2018). Determination of urease biochemical properties of asparagus bean (Vigna unguiculata ssp sesquipedalis L.). IOP Conference Series: Materials Science and Engineering, 349(1). https://doi.org/10.1088/1757-899X/349/1/012073
Zusfahair, Z., Ningsih, D. R., Putri, D., & Fatoni, A. (2018). Partial purification and characterization of urease from black-eyed pea ( Vigna unguiculata ssp unguiculata L . ). Malaysian Journal of Fundamental and Applied Sciences, 14(1), 20–24.
DOI: 10.15408/jkv.v7i2.21768
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