Statistical Optimization of Urease Production by Bacillus B10 Strain Isolated From Kilis Soils

Abstract

In this study, urease producer Bacillus spp. strains were isolated from Kilis soil samples. The specific activities of intracellular urease purified from strains named as B10 and B11 were 1.38±0.07 and 1.22±0.02 µmol/min/mg. Box Behnken design using "Design Expert statistical software" (Design Expert 8.0.7) was applied to increase the urease production of B10 strain. Maximum urease production was observed in a medium containing 5.00% urea, 3.00% sucrose and 2.00% meat extract and the highest specific activity under optimum conditions was calculated as 1.498 ± 0.05 µmol/min/mg.

Keywords

• Bacillus spp.
• Box Behnken
• urease
• Surface Response Methodology

Kilis Topraklarından İzole Edilen Bacillus B10 Suşunun Üreaz Üretiminin İstatiksel Optimizasyonu

Abstract

Bu çalışmada Kilis ili toprak örneklerinden üreaz üretici Bacillus spp. suşları izole edilmiştir. B10 ve B11 olarak isimlendirilen suşlardan saflaştırılan intraselüler üreazın spesifik aktiviteleri 1.38±0.07 ve 1.22±0.02 µmol/min/mg’dır. B10 suşunun üreaz üretimini arttırabilmek için “Design Expert statistical software” (Design Expert 8.0.7)’in kullanıldığı Box Behnken deneme planı uygulanmıştır. Maksimum üreaz üretimi %5.00 üre, %3.00 sükroz ve %2.00 et özütü içeren besi ortamında gözlenmiş ve optimum koşullardaki en yüksek spesifik aktivite 1.498±0.05 µmol/min/mg olarak hesaplanmıştır.

Keywords

• Bacillus spp.,
• Box Behnken
• üreaz
• Yüzey yanıt metodolojisi

References

1. [1] F. Kara, G. Demirel and H. Tumturk, “Immobilization of Urease b Using Chitosan–Alginate and Poly (Acrylamide-Co-Acrylic Acid)/Κ-Carrageenan Supports,” Bioprocess and Biosystems Engineering, vol. 29, no. 3, pp. 207-211, 2006.
2. [2] N. Güleşçi, “Üreaz enziminin magnetik nanopartiküllere farklı ara kollar üzerinden immobilizasyonu ve karakterizasyonu,” Doktora Tezi, Kimya, Çukurova Üniversitesi, Adana, Türkiye, 2013.
3. [3] S. Ciurli, C. Marzadori, S. Benini, S. Deiana and C. Gessa, “Urease from the Soil Bacterium Bacillus pasteurii: Immobilization on Ca-polygalacturonate,” Soil Biology and Biochemistry, vol. 28, no. 6, pp. 811-817, 1996.
4. [4] R.L. Blakeley and B. Zerner, “Jack Bean Urease: The First Nickel Enzyme,” Journal of Molecular Catalysis, vol. 23, no. 3, pp. 263-292, 1984.
5. [5] V. Anitha, K. Abinaya, S. Prakash, A. Seshagiri Rao and B. Vanavil, “Bacillus cereus KLUVAA Mediated Biocement Production Using Hard Water and Urea,” Chemical and Biochemical Engineering Quarterly, vol. 32, no. 2, pp. 257-266, 2018.
6. [6] I. Konieczna, P. Zarnowiec, M. Kwinkowski, B. Kolesinska, J. Fraczyk, Z. Kaminski and W. Kaca, “Bacterial urease and its role in long-lasting human diseases,” Current Protein and Peptide Science, vol. 13, no. 8, pp. 789-806, 2012.
7. [7] D. Örnek Acar, “Üreaz enziminin Ca-alginat üzerine immobilizasyon koşullarının incelenmesi,” Yüksek Lisans Tezi, Kimya, Adnan Menderes Üniversitesi, Aydın, Türkiye, 2009.
8. [8] A. Agarwal, S. Banerje and K. Dev, “Comparative Characterization of Urease Secreting Bacterial and Fungal Isolates from Soil Sample of Farm Fields,” International Journal of Advanced Scientific and Technical Research, vol. 4, no. 5, pp. 138-150, 2015.

9. [9] R. Ramesh, M. Aarthy, M.K. Gowthaman, K. Gabrovska, T. Godjevargova and N.R. Kamini, “Screening and production of a potent extracellular Arthrobacter creatinolyticus urease for determination of heavy metal ions,” Journal of Basic Microbiology, vol. 54, no. 4, pp. 285-295, 2014.
10. [10] B. Sujoy and A. Aparna, “Enzymology, immobilization and applications of urease enzyme,” International Research Journal of Biological Sciences, vol. 2, no. 6, pp. 51-56, 2013.
11. [11] V. Achal, A. Mukherjee and M.S. Reddy, “Characterization of Two Urease-Producing and Calcifying Bacillus spp. Isolated from Cement,” Journal Of Microbiology And Biotechnology, vol. 20, no. 11, pp. 1571-1576, 2010.
12. [12] M.P.E. Berthelot, “Berthelot’s reaction mechanism,” Report de Chimie Applique, 2884, 1859.
13. [13] O.H. Lowry, N.J. Rosebrough, N.J. Farra and R.J. Randall, “Protein measurements with the folin phenol reagent,” Journal of Biological Chemistry, vol. 193, no. 1, pp. 265-275, 1951.
14. [14] M. Mols and T. Abee, “Role of Ureolytic Activity in Bacillus cereus Nitrogen Metabolism and Acid Survival,” Applied and Environmental Microbiology, vol. 74, no. 8, pp. 2370-2378, 2008.
15. [15] D.W. Jeong, B. Lee, H. Lee, K. Jeong, M. Jang, and J.H. Lee, “Urease Characteristics and Phylogenetic Status of Bacillus paralicheniformis,” Journal of Microbiology and Biotechnology, vol. 28, no. 12, pp. 1992-1998, 2018.
16. [16] M. Tepe, S. Arslan, T. Koralay, and N. Mercan Dogan, “Precipitation and Characterization of CaCO3 of Bacillus amyloliquefaciens U17 Strain Producing Urease and Carbonic Anhydrase,” Turkish Journal of Biology, vol. 43, no. 3, pp. 198-208, 2019.
17. [17] J.K. Kim, S.B. Mulrooney and R.P. Hausinge, “Biosynthesis of active Bacillus subtilis urease in the absence of known urease accessory proteins,” Journal Bacteriology, vol. 187, no. 20, pp. 7150-7154, 2015.
18. [18] N.N. Bozbeyoğlu, T.Ş. Candoğan, Ş. Arslan, B. Kabalay, O. Bozkaya, E. Akyol ve N.M. Doğan, “Üreaz ve Karbonik Anhidraz Pozitif Bakteriler Tarafindan Kalsiyum Karbonat Çökelimi,” Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi, c. 26, s. 3, ss. 513-518, 2020.
19. [19] T. Azarno, “Mikrobiyal biyoçimento üretimine yönelik üreaz enzimi üretiminin optimizasyonu,” Yüksek Lisans Tezi, Biyomühendislik, Ege Üniversitesi, İzmir, Türkiye, 2013.
20. [20] A.I. Omoregie, G. Khoshdelnezamihaa, N. Seniana, D.Ek. L. Ong and P.M. Nissom, “Experimental Optimisation of Various Cultural Conditions on Urease Activity for Isolated Sporosarcina pasteurii Strains and Evaluation of Their Biocement Potentials,” Ecological Engineering, vol. 109, no. 2017, pp. 65-75, 2017.
21. [21] V.K. Nathan, J. Vijayan and A. Parvathi, “Optimization of Urease Production by Bacillus halodurans PO15: A Mangrove Bacterium from Poovar mangroves, India,” Marine Life Science and Technology, vol. 2, pp. 194-202, 2020.
22. [22] T. Patil, A. Pendse and K. Aruna, “Optimization of Urease Production by Bacillus megaterium Tara26 Isolated from Marble Quarry Sample and Its Application in Reduction of Water Hardness,” Indian Journal of Applied Research, vol. 10, no. 4, pp. 50-57, 2020.