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Volume: 3, Issue: 5, Sep-Oct, 2015
DOI: 10.7324/JABB.2015.3506

Research Article

Production and Characterization of Alkaline Phosphatase Produced by Bacillus Species

Suganya Kannaiyram, Ravikumar Vedhachalam, Murugan Thanigaimalai

Abstract

Alkaline Phosphatase enzyme was produced from a novel bacterium Bacillus sp. isolated from rocky soil. Enzyme production was carried out in 250 ml of flasks. The crude enzyme was purified by purification, dialyses and chromatography. The enzyme showed 0.28 U/mg1 enzyme activity, 0.31 mg of protein concentration and molecular weight of 54.5 Kda. Also optimum pH and Temperature of the enzyme was pH 10 and 60°C respectively.

Keywords: Bacterial enzyme, Alkaline Phosphatase, Bacillus sp. and SDS-PAGE..

References

1. Holander VP. Acid Phosphatases. In: Enzymes. P.D. Boyer (Ed.) Academic Press, NewYork. 1971; 450-498.

2. Mori S, Okamoto M, Nishibori M, Ichimura M, Sakayama J, Endo H. Purification and characterization of alkaline phosphatase from Bacillus stearothermaphilus. Biotechnol. Applied Biochem. 1999; 29: 235-239.

3. Rina M, Pozidis C, Mavromatis K, Tzanodaskalaki M, Kokkinidis M, Bouriotis V. Alkaline phosphatase from the Antarctic strain TAB5. Properties and psychrophilic adaptations. Europian Journal of Biochemistry. 2000; 267: 1230-1238.

4. Torriani A. Alkaline phosphatase subunit and their dimerization in vivo. Journal Bacteriology. 1968; 96: 1200- 1207.

5. Friedberg I, Avigad G. Some properties of alkaline phosphatase of Ps. flourescens. Europian Journal Biochemistry. 1967; 1: 193-198.

6. Wolfenden R, Spence G. Depression of phosphomonoesterase and phosphodiesterase activities in Aerobacter aerogenes. J. Biochim. Biophys. Acta. 1967; 46: 296-298.

7. Takeda K, Tsugita A. Phosphoesterase of B. subtilis. II crystallization and properties of alkaline phosphatase. Journal of Biochemistry. 1967; 61: 231-241.

8. Michaeliss, Beckwith J. Mechanism of incorporation of cell envelope proteins in Escherichia coli. Annual Review of Microbiology. 1982; 36: 435-465.

9. Hulett FM, Stuckmann K, Spencer DB, Sanopoulou T. Purification and Characterization of the Secreted Alkaline Phosphatase of Bacillus licheniformis MC14: Identification of a Possible Precursor. Journal of General Microbiology. 1986; 132: 2387-2395.

10. Hydrean C, Ghosh A, Nallin M, Ghosh BK. Interrelationship of carbohydrate metabolism and APase synthesis in Bacillus licheniformis 759/C. Journal of Biological Chemistry. 1977; 252: 6806-6812.

11. Glew RH, Heath EC. Studies on the extracellular alkaline phosphatase of Micrococcus sodonensis: Isolation and characterization. J. Biol. Chem. 1971; 246:1556\—1565.

12. Kobori H, Taga N. Extracellular alkaline phosphatase from marine bacteria: purification and properties of extracellular phosphatase from a marine Pseudomonas sp. Can. Journal of Microbiology. 1980; 26:833-838.

13. Nomoto M, Ohsawa M, Wang HL, Chen CC, Yeh KW. Purification and characterization of extracellular alkaline phosphatase from an alkalophilic bacterium. Agric. Biol. Chem. 1988; 52:1643-1647.

14. Prada PD, Jennifer LC, Jean EB. Production of Two Extracellular Alkaline Phosphatases by a Psychrophilic Arthrobacter Strain. Applied and Environmental Microbiology. 1996; 62 (10):3732-3738.

15. Hulett FM, Wangt P, Sussman M, Lee JW. 1985. Proc. Natl. Acad. Sci. USA, Biochemistry, 82, 1035-1039.

16. Mahesh M, Guleria Neha, Rajesh TS, Somashekhar R, Puttaiah ET. Isolation and characterization of extracellular thermostable alkaline phosphatase enzyme from Bacillus spp. International Journal of Applied Biology and Pharmaceutical Technology. 2010; 1(1).

17. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin-Phenol reagents. J. Biol. Chem. 1951; 193: 265-275.

18. Kostadinova S, Marhova M. Purification and properties of alkaline phosphatase from Bacillus cereus second balkan conference on biology special edition/online 2010.

19. Laemmli UK. Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature. 1970; 227: 680-685.

20. Steubing PM. 1993. Isolation of an unknown bacterium from soil. In: Tested studies for laboratory teaching. C. A. Goldman (Ed). 14: 81-114. Proceedings of the 14th Workshop/Conference of the Association for Biology.

21. Sayer PD. Production of alkaline phosphates from E.coli in continuous culture. Applied Microbiology. 1968; 16(2): 326-329.

22. Robert RB, Evan RK. Characterization of a monomeric E.coli alkaline phosphatase formed upon a single amino acid substitution. J. Biol. Chem. 2003; 278: 23497-23501.

23. Hulett FM, Campbell L, Purification and properties of an alkaline phosphate of B. licheniformis. J. Biochem. 1971; 10: 1364-1371.

24. Guimaraes LHS, Terenzi HF, Jorge JA, Leone FA Plizeli MLTM. Extracellur alkaline phosphatase from the filamentous fungus Asperigillus caespitosus: Purification and Biochemical characterization. Folia Microbiology. 2003; 48: 627-632.

25. Junior AB, Guimaraes LHS, Terenzi HF, Jorge JA, Leone FA, Plizeli MLTM. Purification by chemical characterization of thermostable alkaline phosphatases produced by Rhizopus microspores var. rhizopodiformis. Folia Microbiology. 2008; 53(6): 509-516.

26. Bryan LW, Keith W. 1981. General principles of biochemical investigation. Principles and techniques of practical biochemistry. (2nd ed):32-34.

27. Kim EE, Wyckoff HW. Reaction mechanisn of alkaline phosphatase based on crystal structures. Twometal ion catalysis. Journal of Molecular Biology. 1991; 218: 449-464.

28. Goldman S, Hecht K, Eisenberg H, Mevarech M. Extracellular Ca2+-dependent inducible alkaline phosphatase from the extremely halophilic archaeabacterium Haloarcula marismortui. J. Bacteriol. 1990; 172: 7065-7070.

How to cite this article:
Suganya K., Ravikumar V. and Murugan T. Production and Characterization of Alkaline Phosphatase Produced by Bacillus Species. J App Biol Biotech. 2015; 3 (05): 031-034. DOI: 10.7324/JABB.2015.3506

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