Enhanced production and characterization of a highly thermostable alkaline protease from Bacillus sp. P-2
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An obligatory alkalophilic Bacillus sp. P-2, which produced a thermostable alkaline protease was isolated by selective screening from water samples. Protease production at 30 °C in static conditions was highest (66 U/ml) when glucose (1% w/v) was used with combination of yeast extract and peptone (0.25% w/v, each), in the basal medium. Protease production by Bacillus sp. P-2 was suppressed up to 90% when inorganic nitrogen sources were supplemented in the production medium. Among the various agro-byproducts used in different growth systems (solid state, submerged fermentation and biphasic system), wheat bran was found to be the best in terms of maximum enhancement of protease yield as compared to rice bran and sunflower seed cake. The protease was optimally active at pH 9.6, retaining more than 80% of its activity in the pH range of 7–10. The optimum temperature for maximum protease activity was 90 °C. The enzyme was stable at 90 °C for more than 1h and retained 95 and 37% of its activity at 99 °C and 121 °C, respectively, after 1 h. The half-life of protease at 121 °C was 47 min.
- Battaglino, R.A., Huergo, M., Pilosof, A.M.R. & Bartholomai, G.B. 1991 Culture requirements for the production of protease by Aspergillus oryzae in solid state fermentations. Applied Microbiology and Biotechnology 35, 292–296.
- Dhandapani, R. & Vijayaragavan, R. 1994 Production of a thermophilic, extracellular alkaline protease by Bacillus stearothermophilus AP-4. World Journal of Microbiology and Biotechnology 10, 33–35.
- Ferrero, M.A., Cartro, G.R., Abate, C.M., Baigori, M.D. & Sineriz, F. 1996 Thermostable alkaline protease of Bacillus licheniformis MIR 29: isolation, production and characterization. Applied Microbiology and Biotechnology 45, 327–332.
- Gennari, F., Miertus, S., Stredansky, M. & Pizzio, F. 1998 Use of biocatalysts for industrial applications. Genetic Engineering and Biotechnology 3 & 4, 14–23.
- George, S., Raju, V., Krishnan, M.R.V., Subramanian, T.V. & Jayaraman, K. 1995 Production of protease by Bacillus amyloliquefaciens in solid-state fermentation and its application in the unhairing of hides and skins. Process Biochemistry 30, 457–462.
- Gupta, R., Gupta, K., Saxena, R.K. & Khan, S. 1999 Bleach-stable, alkaline protease from Bacillus sp. Biotechnology Letters 21, 135–138.
- Gusek, T.W., Wilson, D.B. & Kinsella, J.E. 1988 Influence of carbon source on production of a heat stable protease from Thermomonospora fusca YX. Applied Microbiology and Biotechnology 28, 80–84.
- Hanlon, G.W., Hodges, N.A. & Russell, A.D. 1982 The influence of glucose, ammonium and magnesium availability on the production of protease and bacitracin by Bacillus licheniformis. Journal of General Microbiology 128, 845–851.
- Kalisz, H.M. 1988 Microbial proteinases. Advances in Biochemical Engineering and Biotechnology 36, 1–65.
- Kole, M.M., Draper, I. & Gerson, D.F. 1988 Production of protease by Bacillus subtilis using simultaneous control of glucose and ammonium concentrations. Journal of Chemical Technology and Biotechnology 41, 197–206.
- Lonsane, B.K., Ghildyal, N.P., Budiatman, S. & Ramakrishna, S.V. 1985 Engineering aspects of solid state fermentation. Enzyme and Microbial Technology 7, 258–265.
- McKeller, R.C. & Cholette, H. 1984 Synthesis of extracellular proteinase by Pseudomonas fluorescens under conditions of limiting carbon, nitrogen and phosphate. Applied and Environmental Microbiology 47, 1224–1227.
- Rahman, R.N.Z.A., Razak, C.N., Ampon, K., Basri, M., Yunus, W.M.Z.W. & Salleh, A.B. 1994 Purification and characterization of a heat-stable alkaline protease from Bacillus stearothermophilus F1. Applied Microbiology and Biotechnology 40, 822–827.
- Sneath, P.H.A. 1986 Endospore-forming gram-positive rods and cocci, Section 13. In Bergey's manual of systematic bacteriology, vol II, eds. Sneath, P.H.A., McNair, N.S. & Sharpe, N.E. pp. 1104–1207. Baltimore: Williams and Wilkins, ISBN 0–683–07893–3.
- Sumandeep, Bhushan, B., Beg, Q.K. & Hoondal, G.S. 1999 Partial purification and characterization of a thermostable alkaline protease of an alkalophilic Bacillus sp. NG-27. Indian Journal of Microbiology 39, 185–189.
- Tyrell, E.A., MacDonald, R.E. & Gerhardt, P. 1958 Biphasic system for growing bacteria in concentrated culture. Journal of Bacteriology 75, 1–4.
- Varela, H., Ferrari, M.D., Belobradjic, L., Vazquez, A. & Loperena, M.L. 1997 Skin unhairing proteases of Bacillus subtilis: production and partial characterization. Biotechnology Letters 19, 755–758.
- Yanga, J.K., Shihb, I.L., Tzengc, Y.M. & Wanga, S.L. 2000 Production and purification of protease from a Bacillus subtilis that can deproteinize crustacean wastes. Enzyme and Microbial Technology 26, 406–413.
- Enhanced production and characterization of a highly thermostable alkaline protease from Bacillus sp. P-2
World Journal of Microbiology and Biotechnology
Volume 17, Issue 2 , pp 125-129
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- Alkaline protease
- Bacillus sp.
- biphasic medium
- solid state
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