Global Advanced Research Journal of Microbiology (GARJM) ISSN: 2315-5116
February 2020 Vol. 9(2): pp. 013-029
Copyright © 2020 Global Advanced Research Journals
Full Length Research Paper
Purification and Characterization of Bacterial Thermoalkalistable Lipase for Application in Bio-detergent Industry
Al-Dhumri, S.A.1 and Bayoumi, R.A.2
1Biology Dept., Khormah University College, Taif University, KSA.
2Botany & Microbiology Dept., Faculty of Science (Boys)، Al-Azhar University, Cairo, Egypt, P.N. 11884. Biology Dept., Khormah University College, Taif University, KSA.
Accepted 13 February, 2020
Lipases with unique substrate speciﬁcity are highly desired in biotechnological applications. Lipases from microbial sources have received heightened attention for an array of industrial applications, and these enzymes have been well exploited in the environmental sector as well. Thermostable lipases occupy a prominent position in aqueous and non-aqueous biocatalysis. The primary goals of this research work are to isolate and identify a lipase producing thermoalkaliphilic Bacillus stearothermophilus- KKSA12 species from oil polluted soil samples collected from Khormah Governorate, KSA. The other purposes of this study are production, partial purification, characterization of lipase activity. The optimum conditions for Bacillus stearothermophilus- KKSA12 hyperthermoalkalistable lipase with slaughter house wastes and tap water, pH value: 11.5; incubation period (hours): 24; incubation temperature (⁰C): 70 ⁰C; substrate concentration (%) (slaughter house wastes):1% ; inoculum size(ml):2ml; incubation conditions: Shaking; best carbon source: Sucrose; Without investigated nitrogen source (Control);Yeast extract concentration(%):1(%); Surfactant: Tween 20; water content : 1 gm slaughter house wastes and 20 ml medium); 100 ml Erlenmeyer flask capacity and 1.25 gm sloid wastes and ZnSO4 (100ml) The batch produced by New Brunswick USA Bioreactor 3000 by submerged fermentation method. The partially purified up to 35.66 % saturation using ammonium sulphate precipitation. One active peak for lipase obtained between 5 to 10 fractions and the fraction number 8 reached the highest specific activity up to 249.28 (U/mg. Protein/ml). Fifteen amino acids were detected. The purified lipase showed the maximum activity at 80⁰C, pH 9.5; at 1.5 % substrate concentration after 60 hours. Chlorine concentration resulted in decreasing the enzyme activity by different ratios depending on the time of enzyme exposure to chlorine. The purified lipase was stable towards strong anionic surfactants. Finally, Bacillus stearothermophilus- KKSA12hyperthermoalkalistable lipase candidate for widely used in food industry, detergent, paper, textile, leather and pharmaceutical industries because of their stability, selectively and substrate specificity for wider industrial applications.
Keywords: Microbial enzymes, Bacterial lipase, Thermoalkaliphilic enzymes, Bio-detergent
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