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Rabab Omran*,Jaafar Anwer Qaddoori


Background: Alkaline phosphatase (ALPase) has vital applications in many aspects of life as molecular biology and genetic engineering applications is used in non-radioactive detection techniques, probing, blotting and sequencing systems and in immunology, diagnosis, linked enzymes in ELISA. Thus it is commercially produced from calf intestine and Escherichia coli. Bacterial enzyme located in periplasmic space of E.coli cells that require costlier extraction method and complex downstream processes. Whereas Bacillus spp. capable to produce Extracellular ALPase which is extracted and purified from crude filtrate and the amount of ALPase is more and easier downstream processes. The present paper amid to isolate new bacterial producer for ALPase regarded to Bacillus sp. and optimize some environmental conditions of enzyme production. Methodology: We used fifteen Bacillus isolates were isolated previously from different soil and waste sources were collected from polluted area, in Advanced Biotechnology and Genetic Engineering Laboratory , College of Science in Babylon University ,Iraq. The production of extracellular ALPase enzymes were screened, in liquid media and the best isolate was selected and identified, subsequently it was grown in different cultural conditions as pH, temperature, incubation periods and aeration and agitation to optimize enzyme production. Results: twelve out of fifteen isolates had capable to produce extracellular ALPase enzymes at variant levels. The best one ( Bacillus sp.I) was selected to produce the enzyme depending for its high value of specific activity of ALPase. After that the environmental conditions for enzyme production were optimize and results revealed that the optimum conditions of extracellular ALPase production in fermentation medium were pH 8.2, 40°C for four days in stand incubator. Conclusion: Most Bacillus isolate produce extracellular ALPase enzymes that encourage more screening of Bacillus isolates for ALPase enzymes production especially that isolated from extreme environment sources as alkaliophilic and thermophilic sources.

Keywords: Extracellular Alkaline Phosphatase, Bacillus, Production, Optimize conditions.

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