Studies on Industrially Significant Haloalkaline Protease from Bacillus sp. JSGT Isolated from Decaying Skin of Tannery

Authors

  • Jaya Sudha Department of Biotechnology, CLRI, Adyar, Chennai-600020, India.
  • V. Ramakrishnan Genetics Division, Central Research Laboratory, Chettinad University, Kelambakkam, Chennai -603103, India.
  • N. Madhusudhanan Department of Pharmacology, Chettinad University, Kelambakkam, Chennai -603103, India.
  • Asit Baran Mandal Chemical Laboratory, CLRI, Chennai-600 020, Tamilnadu, India.
  • Thiyagarajan Gurunathan Department of Biotechnology, CLRI, Adyar, Chennai-600020, India.

Keywords:

Alkaliphiles Bacillus species, Alkaline protease, Decaying organic matter

Abstract

Eight bacterial strains were isolated from collagen layer of decaying skin sample. Three isolates exhibited the prominent zones of clearance on skim milk agar medium at pH 9.5. These isolates were then characterized and identified. One of the haloalkalophilic isolates belonged to the genus Bacillus. Maximum enzyme activity (228.29 ± 1.89 PU/ ml) was found at pH 9 and temperature 37°C in the strain which is designated as Bacillus sp. JSGT. Basic properties such as effects of different temperature, pH, metal ions and inhibitors on protease activity were also studied. Maximum activity was obtained at pH 9 at 55°C. Ca+2 and Mg+2 ions were found to enhance the relative enzyme activity up to 158 and 136% respectively. However, the activity of protease was completely inhibited by phenyl methyl sulfonyl fluoride (PMSF) that showed its serine nature. The results indicated that enzyme produced by Bacillus sp. JSGT is active within broad ranges of temperature and pH. These characteristics render its potential use in leather and detergent industries.

Downloads

Download data is not yet available.

References

Horikoshi, K. (1999). Alkaliphiles: some applications of their products for biotechnology. Microbiol. Mol. Biol. Rev., 63(4): 735–750. https://doi.org/10.1128/MMBR.63.4.735-750.1999.

Demirjian, D.C., Morís-Varas, F. & Cassidy, C.S. (2001). Enzymes from extremophiles. Curr. Opin. Chem. Biol., 5(2): 144–151. https://doi.org/10.1016/s1367-5931(00)00183-6.

Nilegaonkar, S.S., Kanekar, P.P., Sarnaik, S.S. & Kelkar, A.S. (2002). Production, isolation and characterization of extracellular protease of an alkaliphilic strain of Arthrobacter ramosus, MCM B-351 isolated from the alkaline lake of Lonar, India. World J. Microbiol. Biotechnol., 18(8): 785–789. https://doi.org/10.1023/A:1020481126362.

Kanekar, P.P., Nilegaonkar, S.S., Sarnaik, S.S. & Kelkar, A.S. (2002). Optimization of protease activity of alkaliphilic bacteria isolated from an alkaline lake in India. Bioresour. Technol., 85(1): 87–93. https://doi.org/10.1016/s0960-8524(02)00018-4.

Ulukanli, Z. & Diğrak, M. (2002). Alkaliphilic micro-organisms and habitats. Turk. J. Biol., 26: 181–191.

Denizci, A.A., Kazan, D., Abeln, E.C. & Erarslan, A. (2004). Newly isolated Bacillus clausii GMBAE 42: an alkaline protease producer capable to grow under higly alkaline conditions. J. Appl. Microbiol., 96(2): 320–327. https://doi.org/10.1046/j.1365-2672.2003.02153.x.

Naidu, K.S.B. & Devi, K.L. (2005). Optimization of thermostable alkaline protease production from species of Bacillus using rice bran. Afr. J. Biotechnol., 4(7): 724–726. https://doi.org/10.5897/AJB2005.000-3132.

Gupta, R., Beg, Q.K. & Lorenz, P. (2002). Bacterial alkaline proteases: molecular approaches and industrial applications. Appl. Microbiol. Biotechnol., 59(1): 15–32. https://doi.org/10.1007/s00253-002-0975-y.

Kumar, C.G. & Takagi, H. (1999). Microbial alkaline proteases: from a bioindustrial viewpoint. Biotechnol. Adv., 17(7): 561–594. https://doi.org/10.1016/s0734-9750(99)00027-0.

Holt, J.G., Krieg, N.R., Sneath, P.H.A., Stately, J.T. & St Williams (1994). Bergey’s Manual of Determinative Bacteriology, 9th Ed., Williams and Wilkins, Baltimore, pp. 787.

Forbes, B.A., Sahm, D.F. & Weissfeld, A.S. (2002). Bacillus and Similar Organisms. In: Bailey & Scott's diagnostic microbiology. 11th Ed., St. Louis: Mosby. pp. 317-324.

Hagihara, B. (1958). Bacterial and mold proteases. In: Boyer, P.D., Lardy, H. & Myrbäck, K. (eds.), The enzymes, Vol. 4, Academic Press Inc, NY, pp. 193-213.

Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem., 193(1): 265–275. https://doi.org/10.1016/S0021-9258(19)52451-6.

Heyrman, J., Vanparys, B., Logan, N.A., Balcaen, A., Rodríguez-Díaz, M., Felske, A. & De Vos, P. (2004). Bacillus novalis sp. nov., Bacillus vireti sp. nov., Bacillus soli sp. nov., Bacillus bataviensis sp. nov. and Bacillus drentensis sp. nov., from the Drentse A grasslands. Int. J. Syst. Evol. Microbiol., 54: 47–57. https://doi.org/10.1099/ijs.0.02723-0.

Aslım, B., Yüksekdağ, Z. & Beyatlı, Y. (2002). Determination of PHB growth quantities of certain Bacillus species isolated from soil. Turkish Electron. J. Biotechnol. (Special Issue), 1: 24-30.

Bach, H.J. & Munch, J.C. (2000). Identification of bacterial sources of soil peptidases. Biol. Fertil. Soils, 31(3): 219–224. https://doi.org/10.1007/s003740050648.

Kumar, C.G., Malik, R.K. & Tiwari, M.P. (1998). Novel enzyme based detergents: an Indian perspective. Curr. Sci., 75: 1312–1318.

Pastor, M.D., Lorda, G.S. & Balatti, A. (2001). Protease obtention using Bacillus subtilis 3411 and amaranth seed meal medium at different aeration rates. Braz. J. Microbiol., 32: 6–9. https://doi.org/10.1590/S1517-83822001000100002.

Moradian, F., Khajeh, K., Naderi-Manesh, H., Ahmadvand, R., Sajedi, R.H. & Sadeghizadeh, M. (2006). Thiol-dependent serine alkaline proteases from Bacillus sp. HR-08 and KR-8102: isolation, production, and characterization. Appl. Biochem. Biotechnol., 134(1): 77–87. https://doi.org/10.1385/abab:134:1:77.

Joo, H.S., Kumar, C.G., Park, G.C., Paik, S.R. & Chang, C.S. (2003). Oxidant and SDS-stable alkaline protease from Bacillus clausii I-52: production and some properties. J. Appl. Microbiol., 95(2): 267–272. https://doi.org/10.1046/j.1365-2672.2003.01982.x.

Horton, H.R., Moran, L.A., Ochs, R.S., Rawn, J.D. & Scrimgeour, K.G. (1996). Properties of enzymes. In: Principles of Biochemistry. 2nd edition, Prentice Hall: Upper Saddle River, NJ, pp. 119-146.

Jeong, Y., Wei, C.I., Preston, J.F. & Marshall, M.R. (2000). Purification and characterization of proteases from hepatopancreas of Crawfish (Procambarvs clarkii). J. Food Biochem., 24(4): 311–332. https://doi.org/10.1111/j.1745-4514.2000.tb00703.x.

Adinarayana, K., Ellaiah, P. & Prasad, D.S. (2003). Purification and partial characterization of thermostable serine alkaline protease from a newly isolatedBacillus subtilis PE-11. AAPS PharmSciTech, 4(4): 440–448. https://doi.org/10.1208/pt040456.

Yossan, S., Reungsang, A. & Yasuda, M. (2006). Purification and characterization of alkaline protease from Bacillus megaterium isolated from Thai fish sauce fermentation process. ScienceAsia, 32(4): 377–383. http://dx.doi.org/10.2306/scienceasia1513-1874.2006.32.377.

Johnvesly, B. & Naik, G.R. (2001). Studies on production of thermostable alkaline protease from thermophilic and alkaliphilic Bacillus sp. JB-99 in a chemically defined medium. Process Biochem., 37(2): 139–144. https://doi.org/10.1016/S0032-9592(01)00191-1.

Donaghy, J.A. & McKay, A.M. (1993). Production and properties of an alkaline protease by Aureobasidium pullulans. J. Appl. Bacteriol., 74(6): 662–666. https://doi.org/10.1111/j.1365-2672.1993.tb05200.x.

Downloads

Abstract views: 33 / PDF downloads: 23

Published

2010-07-01

How to Cite

Sudha, J., Ramakrishnan, V., Madhusudhanan, N., Mandal, A. B., & Gurunathan, T. (2010). Studies on Industrially Significant Haloalkaline Protease from Bacillus sp. JSGT Isolated from Decaying Skin of Tannery. Advances in BioScience, 1(1), 46–51. Retrieved from https://journals.sospublication.co.in/ab/article/view/6

Issue

Section

Articles