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IJSTR >> Volume 3- Issue 12, December 2014 Edition

International Journal of Scientific & Technology Research  
International Journal of Scientific & Technology Research

Website: http://www.ijstr.org

ISSN 2277-8616

Some Properties Of Thermo-Stable Α -Amylase Of Four Isolates Of Bacillus Licheniformis

[Full Text]



Elham S. Dawood, Ibrahim S. A., Sohair A. Abdelwahab



Keywards: Bacillus licheniformis, thermo-stable amylase, α-Amylases, partial purification ,DEAD Sephadex.



Abstract: Four isolates of Bacillus licheniformis producing thermo-stable amylase enzymes were previously isolated from different Sudanese soils and designated as B. licheniformis SUDK1, SUDK2, SUDK4 and SUDO. The enzyme was partially purified using DEAD Sephadex A 25 gel filtration and then were identified using thin layer chromatography. Optimum pH and temperature for enzyme activity were determined. The effect of thermal stability was detected at two storage temperatures (-20°Cand 4 °C) for 24 weeks. The results showed that the activity of the partially purified enzymes increased up to 16 -18 folds. The amylase enzymes were found to hydrolyze starch forming various maltooligosaccharides, such as dextrins and maltose as major products so they were identified as α- amylases. Optimum temperatures for enzyme activity was obtained at 60 and 70 °C. All enzymes were stable between pH 6.0-9.0 with optimum pH 7.0 . The enzymes were stable and retained nearly all of their initial activities at -20 °C till the end of 24th week. At 4 °C they lost less than 60% of their initial activities at the 8th week. The Km values were 1.25 – 2.0 mg/ml.



[1] S. Yandri, S. Tati, and H. Sutopo,. Purification andCharacterization of Extracellular α-Amilase Enzyme from Locale Bacteria Isolate Bacillus subtilis ITBCCB1. Europ. J. Sci. Res. (2010), 39(1), 64-74.

[2] G.Dey, Palit S, Banerjee R, Maiti BR). Purification and characterization of maltooligosaccharide-forming amylase from Bacillus circulans GRS313 J. Ind. Microbiol. Biotechnol. (2002), 28, 193–200.

[3] EB. Messaoud, MB Ali, N. Elleuch, NF Masmoudi. and S. Bejar . Purification and properties of a maltoheptaose- and maltohexaoseforming amylase produced by Bacillus subtilis US116. Enzyme Microb. Technol.(2004), 34, 662–666.

[4] S.O. Hashim; O. Delgado; R. Hatti-Kaul; F.J. Mulaa; B. Mattiasson Starch hydrolysing Bacillus halodurans isolates from a Kenyan soda lake. Biotechnol. Lett. (2004), 26, 823-828.

[5] Kathiresan and Manivannan, α- Amylase production by Pencillium fellutanum isolated from mangrove rhizosphere soil. Afr. J. Biotech. ( 2006), 5(10), 829-832.

[6] Z. Chi; G. Liu; F. Wang; L. Ju and T. Zhang Saccharomycopsis fibuligera and its applications in biotechnology. Biotechnol Adv. (2009), 27, 423- 431.

[7] A. Gupta, VK. Gupta, DR. Modi and LP. Yadava. Production and characterization of α-amylase from Aspergillus niger. Biotechnol 2008, 7(3):551.
[8] X. D. Liu, Y. Xu, . A novel raw starch digesting a-amylase from a newly isolated Bacillus sp. YX-1: Purification and characterization. (2008), 88(10) ,3415-3420.

[9] R.K. Saxena; K. Dutt; L. Agarwal and P. Nayyar. A highly thermostable and alkaline amylase from a Bacillus sp. PN5. Bioresour Technol (2007),98, 260-265.

[10] A. Burhan, U. Nisa, C. Gokhan, C. Omer,A. Ashabil and G. Osman. Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus sp. isolate ANT-6. Proc. Bioch., (2003) 38, 1397-1403.

[11] Z. Konsula and M. liakopoulou-Kyriakides, “Hydrolysis of starches by the action of α-amylase from Bacillus subtilis”. Proc Biochem., (2004) ,39, pp.1745-1749.

[12] G. L. Miller Use of dinitrosalicylic acid reagent for determination of A novel raw starch digesting a-amylase from a newly isolated Bacillus sp. YX-1: (1972). Purification and characterization.

[13] M.M. Bradford .A rapid and sensitive method for the quantities of microgram quantities of protein – dye binding. Analytical Biochemistry 72, 248- 254 (1976).

[14] W. E. Trevelyan, D. P. Procter and, J. S. Harrison. Detection of sugar on paper chronotograms. Nature, London 166, 444-448 (1950).

[15] M. Adeyanju; F. Agboola; B. Omafuvbe; O. Oyefuga and O.Adebawo. A Thermostable α- Amylase from Bacillus licheniformis Isolated from Cassava Steep Water. Biotech., (2007), 6(4), 473-480.

[16] F. Al-Quadan, H. Akel and R. Natshi. Characteristics of a Novel Highly Thermostable and Extremely Thermophilic Alkalitolerant Amylase from Hyperthermophilic Bacillus Strain HUTBS71. OnLine J. of Biol. Sci. (2009) 9 (3), 67-74.

[17] S. kumar , M. Subhosh, K. Chandra , V. Mallaiah, V. Sreenivasulu and Yong-Lark Choi .Purification and Characterization of Highly Thermostable α-amylase from Thermophilic Alicyclobacillus acidocaldarius Biotechnology and Biopr. Eng. (2010),15.

[18] M. Atiyeh, H. Reza, Sajedi, R. Mehdi and Vahab. Characterization of an α-amylase with broad temperature activity from an acid-neutralizing Bacillus cereus strain. Iran. J. Biotech., ( 2010), 8(2) 154 165.

[19] P. Deb, S. Ahmad S., Mohsina K ., Kumar P. Sarker and Abu Sayem S . Production and partial characterization of extracellular amylase enzyme from Bacillus amyloliquefaciens P-00 (2013) http://www.springerplus.com/content/2/1/154

[20] V. Raquel et al .PROPERTIES of an amylase from thermophili Bacillus sp. Braz. J. Microb.( 2008) , 39,102-107