2014, Issue 2, Volume 6

CHARACTERIZATION OF THERMOPHILIC AMYLASE FROM AN OBLIGATE THERMOPHILE, THERMOACTINOMYCES VULGARIS

Archana Singh and Ved Pal Singh*

Applied Microbiology and Biotechnology Laboratory, Department of Botany, University of Delhi, Delhi- 110 007, India

* E-mail: vpsingh_biology@rediffmail.com

Abstract: Amylase finds a wide range of applications in starch industries, i.e., baking, brewing, distillery. The wild-type (1227) and mutant strains (1261 and 1286) of Thermoactinomyces vulgaris were screened for the production of amylase using 1% soluble starch. The maximum production of amylase was observed after 12 h of incubation at 50ºC in wild-type strain 1227 of T. vulgaris. The amylase was found to be thermophilic, exhibiting its optimal activity at 75ºC and at pH 6.0 in this obligate thermophile; and it preferred soluble starch as its substrate. Among the metal ions tested, Mn2+ was most stimulatory, while Hg2+ was most inhibitory to the activity of amylase. Thus, T. vulgaris amylase is a thermophilic metalloenzyme, requiring Mn2+ for its high-temperature catalysis, which can be exploited for amylase-based   industries of diverse interests.

Keywords: Amylase, Metalloenzyme, Thermoactinomyces vulgaris, Thermophilic amylase

REFERENCES

Asgher, M., Asad, M.J., Rahman, S.U. and Legge, R.L. (2007). A thermostable α-amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. J. Food Eng. 79, 950-955.

Bernfeld, P. (1955). Amylases α/β. Methods in Enzymology 1: 149-158.

Bradford, M. (1976). A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254.

Burhan, A., Nisa, U. and Gokhan, C. (2003). Enzymatic properties of a novel thermostable, thermophilic alkaline and chelator resitant amylase from an alkaliphilic Bacillus sp. isolate ANT-6. Process Biochem. 38: 1397-1403.

Glymph, J.L. and Stutzenberger, F.J. (1977). Production, purification and characterization of α-amylase from Thermomonospora curvata. Appl. Env. Microbiol. 34: 391-397.

Hagihara, H., Igarashi, K., Hayashi, Y., Endo, K., Kitayama, K.J., Kawai, S., Kobayashi, T. and Ito, S. (2001). Novel α-amylase that is highly resistant to chelating reagents and chemical oxidants from the alkaliphilic Bacillus isolate KSM-K38. App. Environ. Microbiol. 67: 1744-1750.

Hopwood, D.A. and Wright, H.M. (1972). Transformation in Thermoactinomyces vulgaris. J. Gen. Microbiol. 71: 383-393.

Kelly, C.T., McTigue, M.A., Doyle, E.M. and Fogarty, W.M. (1995). The raw starch degrading alkaline amylase of Bacillus sp. IMD370. J. Ind. Microbiol. Biotechnol. 15: 446-448.

Mamo, G., Gashe, B.A. and Gessesse, A. (1999). A highly thermostable amylase from a newly isolated thermophilic Bacillus sp. WN11. J. App. Microbiol. 86: 557-560

Rao, M.B., Tanksale, A.M., Gathe, M.S. and Deshpande, V.V. (1998). Molecular and biotechnological aspects of microbial proteases. Microbiol. Mol. Biol. Reviews 62: 597-635.

Shanmughapriya, S., Wran, G.S., Selvin, J., Gandhimathi, R., Baskar, T.B., Manilal, A. and Sujith, S. (2009). Optimization, production, and partial characterization of an alkalophilic amylase produced by sponge associated marine bacterium Halobacterium salinarum MMDO47. Biotechnol. Bioprocess Eng. 14: 67-75.

Sidhu, G.S., Shrma, P., Chakrabarti, T. and Gupta, J.K. (1997). Strain improvement for the production of a thermostable α-amylase. Enz. Microbiol. Biotechnol. 21: 525-530.

Singh, V.P. (1980). Investigations on the phosphatases of Thermoactinomyces vulgaris. Ph.D. Thesis, University of Delhi, India.

Singleton, R. and Amelunxen, R.E. (1973). Proteins from thermophilic microorganisms. Bacteriol. Rev. 37: 320-342.

Sinha, U. and Singh, V.P. (1980). Phosphate utilization and constitutive synthesis of phosphatases in Thermoactinomyces vulgaris Tsilinsky. Biochem. J. 190: 457-460.

Siva Sakthi, S., Kanchana, D. and Saranraj, P. and Usharani, G.  (2012).Evaluation of amylase activity of the amylolytic fungi Aspergillus niger usingcassava as substrate. International Journal ofApplied Microbiology Sciences 1(1): 24-34.

Sivaramakrishnan, S., Gangadharan, D., Nampoothiri, K.M., Soccol, C.R. and Pandey, A. (2006). α-Amylases from microbial sources-an overview on recent developments. Food Technol. Biotechnol. 44: 173-184.

Zuber, H. (1976). Enzymes and proteins from thermophilic microorganisms. Birkhaeuser Veralag, Basel and Stuttgart. pp 445.