Isolation of a Native Bacillus licheniformis Amylase Producer from Hot Source of Semnan

Subject Areas : Environmental Microbiology

Majid Moghbeli ¹ , Hamid Noshiri ²

1 - Department of Microbiology, Damghan Branch, Islamic Azad University, Damghan, Iran
2 - Department of Microbiology, Qom Branch, Islamic Azad University, Qom, Iran

Received: 2013-09-16 Accepted : 2013-10-17 Published : 2009-10-01

Keywords: Bacillus licheniformis, Amylase, Hot spring,

Abstract :

Background and Objectives: Amylase is one of the important industrial enzymes, which is used in the food industry, detergent production, glucose, textile, and paper making. Bacillus spp. are considered as a major amylase and protease producer microorganisms. The main purpose for the study is isolating amylase positive thermophile Bacillus Spp. from hot sources of Semnan province. Material and Methods: Soil, fertilizer and water samples were collected from environment and the hot springs. After dilution preparation, samples were cultured on starch agar medium and then, potential of amylase producing was measured. Alpha-amylase enzyme activity of isolates was measured using di-nitro salicylic acid (DNS) method. Results: Among 10 amylase producing isolated bacteria, two samples showed the highest amylase activity, 104.5u/ml and 61.2u/ml respectively. Although both the samples were diagnosed as Bacillus spp. one of them was belong to mesophile species while another one was detected as thermophile Bacillus. The thermophile Bacillus has been detected as a member of B. licheniformis according to Bergey’s Manual of Systematic Bacteriology. Conclusion: Due to high consumption of amylase enzymes and lack of producing the enzymes in Iran, The isolated Bacillus licheniformis in this study, with 61.2 U product per mL, can be introduced as candidate for produce the favorite amount of the enzyme after proper genetic manipulations.

References:

1) Boyer, E.W., and M. B. Ingle. Extracellular alkaline amylase from a Bacillus species. J. Bacteriol, 1972; 110:992-1000.
2) Ivanova, V., E. Dobreva, E. Emanuilova. Purification and characterization of thermostable alpha-amylase from Bacillus licheniformis. J. Biotechnol, 1993; 28, 277-289.
3) Upadek, H., and B. Kottwitz. Application of amylases in detergents. In J. H. van Ee, O. Misset, and E. J. Baas (ed.), Enzymes in detergency. Marcel Dekker, Inc., New York. 1997; p. 203-212
4) Van der Laan, J. M. May  Novel amylolytic enzymes derived from the B. licheniformis, having improved characteristics.International patent, 1995; WO 95/35382.
5) Van der Maarel, M.J.E.C., Van der Veen, B., Uitdehaag J.C.M., and H. Leemhuis. Properties and applications of starch-converting enzymes of the alfa amylase family (Reviev article). J. Bacteriol, 2002; 94: 137-155
6) Shafiei M, Ziaee AA, Amoozegar MA., Purification and characterization of an organic-sol-vent-tolerant halophilic α-amylase from the moderately halophilic Nesterenkonia sp. strain F. J Ind Microbiol Biotechnol, 2011; 38(2): 275-81
7) Michelin M, Silva TM, Benassi VM, Peixoto-Nogueira SC, Moraes LA, Leão JM, Jorge JA, Terenzi HF, Polizeli Mde L., Purification and characterization of a thermostable α-amylase produced by the fungus Paecilomyces variotii. Carbohydr Res., 2010; 2 (16): 2348-53.
8) Nakajima, R., T. Imanaka, and S. Aiba. Comparison of amino acid sequences of eleven different α-amylase. Appl. Microbiol. Biotechnol, 1986; 23: 355-60.
9) Nakajima, R., T. Imanaka, and S. Aiba. Nucleotide sequence of the Bacillus stearothermophilus α-amylase gene. J. Bacteriol, 1985; 163: 401-6.
10) Yamaguchi R, Tokunaga H, Ishibashi M, Arakawa T, Tokunaga M., Salt-dependent thermo-reversible α-amylase: cloning and characterization of halophilic α-amylase from moderately halophilic bacterium, Kocuria varians. Appl Microbiol Biotechnol, 2011;
89(3): 673-84
11) Liu XD, Xu Y,A novel raw starch digesting alpha-amylase from a newly isolated Bacillus sp. YX-1: purification and characterization. Bioresour Technol. 2008, 99(10):4315-20.
12) Harley M, Prescott J., Laboratory exercise in microbiology, 2002; Fifth edition. The McGraw Hill,
13) Stefanova, M., E. Emanuilova. Characterization of a thermostable alphaamylase from Bacillus brevis. Eur. J. Biochem., 1992; 207, 345-9.
14) Brenner D.J., Krieg N.R., Statey J.T., Bergey’s manual systematic bacteriology, vol 2, part B, Springer.