IDENTIFICATION OF THERMOPHILIC BACTERIA BACILLUS BADIUS W.IIISRNA_2.1 IS A POTENTIALLY NOVEL STRAIN FROM MATAUMPANA HOT SPRINGS, BUTON, INDONESIA, AS PRODUCERS OF URICASE ENZYME

Authors

  • Sarni Politeknik Baubau, Sulawesi Tenggara, Indonesia
  • Hasnah Natsir Chemistry Department, Hasanuddin University, Makassar, South Sulawesi, Indonesia
  • Abdul Wahid Wahab Chemistry Department, Hasanuddin University, Makassar, South Sulawesi, Indonesia
  • Abdul Karim Chemistry Department, Hasanuddin University, Makassar, South Sulawesi, Indonesia
  • Nur Umriani Permatasari Chemistry Department, Hasanuddin University, Makassar, South Sulawesi, Indonesia
  • Wahyudin Rauf Chemistry Department, Hasanuddin University, Makassar, South Sulawesi, Indonesia
  • Evi Mustiqawati oliteknik Baubau, Sulawesi Tenggara, Indonesia

DOI:

https://doi.org/10.11113/jurnalteknologi.v88.23713

Keywords:

Thermophilic bacteria, uricase, uric acid, Bacillus badius, mataumpana hot springs

Abstract

Thermophilic bacteria are unique microorganisms because they can survive and thrive in high temperatures and extreme environments such as hot springs. Microorganisms that survive in environment like this contain active compounds, one of which is the enzyme uricase. Uricase is an enzyme that plays an important role in the nitrogen metabolism pathway to catalyze oxidation of uric acid into allantoin, CO2 and H2O2 which are water soluble easily. This study aims to identify thermophilic bacterial species producers of uricase enzyme isolated from Mataumpana hot spring water samples and determine the effect of incubation time on bacterial growth and uricase enzyme production. Bacterial identification includes morphological and biochemical tests as well as species-level molecular tests using 16S rRNA, then determining the effect of incubation time on bacterial growth and uricase enzyme production. The results of the analysis showed that the uricase-producing thermophilic bacterial isolate W.IIISRNa_2.1 strain was identified as coming from the gram-positive Bacillus genus and had a very close relationship with the Bacillus badius species with a similarity of 98.97%, so the isolate was named Bacillus badius W.IIISRNa_2.1. This bacterium has an optimal optical density (OD) at 42 hours of incubation of 0.962, which is different from the incubation time for production of uricase, was 12 hours with an enzyme activity of 0.4928 U/ml and a protein concentration of 4.7715 mg/ml. 

References

Sarni, S., A. W. Wahab, H. Natsir, N. La Nafie, and A. R. Arif. 2023. Production of Crude Uricase Enzyme by Novel Bacillus altitudinis Strain W.IIISRNs_1.1 from the Hot Spring of Mataumpana, Buton Regency, Southeast Sulawesi. Egyptian Journal of Chemistry. 66(12): 115–126. https://doi.org/10.21608/ejchem.2023.150355.6511.

Sabaria, E., Y. Yasmin, Y. S. Ismail, M. A. Bessania, I. Putri, and L. Fitri. 2024. Characterization of Thermophilic Bacteria from Ie Seum Hot Springs, Aceh Besar, Indonesia as Producers of Protease Enzyme. Biodiversitas. 25(5): 1867–1874. https://doi.org/10.13057/biodiv/d250502.

Devi, R., and C. S. Pundir. 2014. Construction and Application of an Amperometric Uric Acid Biosensor Based on Covalent Immobilization of Uricase on Iron Oxide Nanoparticles/Chitosan-g-Polyaniline Composite Film Electrodeposited on Pt Electrode. Sensors and Actuators B: Chemical. 193: 608–615. https://doi.org/10.1016/j.snb.2013.12.010.

Fukuda, T., H. Muguruma, H. Iwasa, T. Tanaka, A. Hiratsuka, T. Shimizu, K. Tsuji, and T. Kishimoto. 2020. Electrochemical Determination of Uric Acid in Urine and Serum with Uricase/Carbon Nanotube/Carboxymethylcellulose Electrode. Analytical Biochemistry. 590: 113533. https://doi.org/10.1016/j.ab.2019.113533.

Khade, S. M., S. K. Srivastava, K. Kumar, K. Sharma, A. Goyal, and A. D. Tripathi. 2018. Optimization of Clinical Uricase Production by Bacillus cereus under Submerged Fermentation, Its Purification and Structure Characterization. Process Biochemistry. 75: 49–58. https://doi.org/10.1016/j.procbio.2018.09.010.

Nanda, P., and P. E. Jagadeesh Babu. 2014. Isolation, Screening and Production Studies of Uricase Producing Bacteria from Poultry Sources. Preparative Biochemistry and Biotechnology. 44(8): 811–821. https://doi.org/10.1080/10826068.2013.867875.

Atty, F., and J. Joseph. 2016. Isolation and Identification of Uric Acid Degrading Bacteria, Optimization of Uricase Production and Purification of Uricase Enzyme. International Journal of Advanced Research. 4(12): 2732–2742. https://doi.org/10.21474/ijar01/2702.

Khade, S., S. K. Srivastava, and A. D. Tripathi. 2016. Production of Clinically Efficient Uricase Enzyme Induced from Different Strains of Pseudomonas aeruginosa under Submerged Fermentations and Their Kinetic Properties. Biocatalysis and Agricultural Biotechnology. 8: 139–145. https://doi.org/10.1016/j.bcab.2016.09.005.

Pierzynowska, K., A. Deshpande, N. Mosiichuk, R. Terkeltaub, P. Szczurek, E. Salido, S. Pierzynowski, and D. Grujic. 2020. Oral Treatment with an Engineered Uricase, ALLN-346, Reduces Hyperuricemia, and Uricosuria in Urate Oxidase-Deficient Mice. Frontiers in Medicine. 7: 569215. https://doi.org/10.3389/fmed.2020.569215.

Roman, Y. M. 2023. The Role of Uric Acid in Human Health: Insights from the Uricase Gene. Journal of Personalized Medicine. 13(9): 1409. https://doi.org/10.3390/jpm13091409.

Omar, M. N., A. B. Salleh, H. N. Lim, and A. Ahmad Tajudin. 2016. Electrochemical Detection of Uric Acid via Uricase-Immobilized Graphene Oxide. Analytical Biochemistry. 509: 135–141. https://doi.org/10.1016/j.ab.2016.06.030.

Tork, S. E., M. M. Aly, and S. Q. Al-Fattani. 2020. A New Uricase from Bacillus cereus SKIII: Characterization, Gene Identification and Genetic Improvement. International Journal of Biological Macromolecules. 165: 3135–3144. https://doi.org/10.1016/j.ijbiomac.2020.10.183.

Handayani, I., T. Utami, C. Hidayat, and E. S. Rahayu. 2018. Screening of Lactic Acid Bacteria Producing Uricase and Stability Assessment in Simulated Gastrointestinal Conditions. International Food Research Journal. 25(4): 1661–1667.

Sarni, S., H. Natsir, N. La Nafie, and A. W. Wahab. 2023. Screening and Identification of Thermophilic Uricase Bacteria from the Mataumpana Hot Spring, Buton Regency, Southeast Sulawesi. AIP Conference Proceedings. 2719: 030023.

Jirakunakorn, R., S. Khumngern, J. Choosang, P. Thavarungkul, P. Kanatharana, and A. Numnuam. 2020. Uric Acid Enzyme Biosensor Based on a Screen-Printed Electrode Coated with Prussian Blue and Modified with Chitosan-Graphene Composite Cryogel. Microchemical Journal. 154: 104624. https://doi.org/10.1016/j.microc.2020.104624.

Tvorynska, S., J. Barek, and B. Josypčuk. 2021. Flow Amperometric Uric Acid Biosensors Based on Different Enzymatic Mini-Reactors: A Comparative Study of Uricase Immobilization. Sensors and Actuators B: Chemical. 344: 130252. https://doi.org/10.1016/j.snb.2021.130252.

Pustake, S. O., P. K. Bhagwat, and P. B. Dandge. 2019. Statistical Media Optimization for the Production of Clinical Uricase from Bacillus subtilis Strain SP6. Heliyon. 5(5): e01756. https://doi.org/10.1016/j.heliyon.2019.e01756.

Song, P., X. Zhang, S. Wang, W. Xu, F. Wang, R. Fu, and F. Wang. 2023. Microbial Proteases and Their Applications. Frontiers in Microbiology. 14: 1236368. https://doi.org/10.3389/fmicb.2023.1236368.

Badoei-Dalfard, A., M. Shaban, and Z. Karami. 2019. Characterization, Antimicrobial, and Antioxidant Activities of Silver Nanoparticles Synthesized by Uricase from Alcaligenes faecalis GH3. Biocatalysis and Agricultural Biotechnology. 20: 101257. https://doi.org/10.1016/j.bcab.2019.101257.

Khade, S. M., S. K. Srivastava, K. Kumar, K. Sharma, A. Goyal, and A. D. Tripathi. 2018. Optimization of Clinical Uricase Production by Bacillus cereus under Submerged Fermentation, Its Purification and Structure Characterization. Process Biochemistry. 75: 49–58. https://doi.org/10.1016/j.procbio.2018.09.010.

El-Naggar, N. E. A. 2015. Isolation, Screening and Identification of Actinobacteria with Uricase Activity: Statistical Optimization of Fermentation Conditions for Improved Production of Uricase by Streptomyces rochei NEAE-25. International Journal of Pharmacology. 11(7): 644–658. https://doi.org/10.3923/ijp.2015.644.658.

Poovizh, T., P. Gajalakshmi, and S. Jayalakshmi. 2014. Production of Uricase, a Therapeutic Enzyme from Pseudomonas putida Isolated from Poultry Waste. International Journal of Advanced Research. 2(1): 34–40.

Yazdi, M. T., G. Zarrini, E. Mohit, M. A. Faramarzi, N. Setayesh, N. Sedighi, and F. A. Mohseni. 2006. Mucor hiemalis: A New Source for Uricase Production. World Journal of Microbiology and Biotechnology. 22(4): 325–330. https://doi.org/10.1007/s11274-005-9030-3.

Lee, N. S. I. S., H. M. Khosravi, N. Ibrahim, and S. Shahir. 2015. Isolation, Partial Purification and Characterization of Thermophilic Uricase from Pseudomonas otitidis Strain SN4. Malaysian Journal of Microbiology. 11(4): 352–357.

Sarni, S., H. Natsir, and S. Dali. 2016. Production and Characterization Chitosanase of Sponge Symbiont Bacteria Klebsiella sp. to Hydrolyze Chitosan Be Chitooligosaccarides. Marina Chimica Acta International Journal. 17(1).

Iswantini, D., N. Nurhidayat, Trivadila, and A. Nurjayati. 2011. Penentuan Kinetika Urikase Dari Sel Bacillus subtilis, B. megaterium, dan B. cereus. Jurnal Ilmu Pertanian Indonesia. 16(2): 112–118.

Hemraj, V., and D. G. A. Sharma. 2013. A Review on Commonly Used Biochemical Test for Bacteria. Innovare Journal of Life Science. 1(1): 221–230. https://doi.org/10.1109/ICDM.2013.109.

Hall, B. G. 2013. Building Phylogenetic Trees from Molecular Data with MEGA. Molecular Biology and Evolution. 30(5): 1229–1235. https://doi.org/10.1093/molbev/mst012.

Ravichandran, R., S. Hemaasri, S. S. Cameotra, and N. S. Jayaprakash. 2015. Purification and Characterization of an Extracellular Uricase from a New Isolate of Sphingobacterium thalpophilum (VITPCB5). Protein Expression and Purification. 114: 136–142. https://doi.org/10.1016/j.pep.2015.06.017.

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Published

2026-06-16

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Science and Engineering