Issa, H., Abou-Dobara, M., El-Sayed, A., El-Bana, M. (2024). Optimization, purification and characterization of extracellular lipase produced by Serratia marcescens EGHK-19. Egyptian Journal of Botany, 64(4), 107-119. doi: 10.21608/ejbo.2024.244359.2545
Heba Kamal Issa; Mohammed Ismaeil Abou-Dobara; Ahmed El-Sayed; Magdy Ibrahim El-Bana. "Optimization, purification and characterization of extracellular lipase produced by Serratia marcescens EGHK-19". Egyptian Journal of Botany, 64, 4, 2024, 107-119. doi: 10.21608/ejbo.2024.244359.2545
Issa, H., Abou-Dobara, M., El-Sayed, A., El-Bana, M. (2024). 'Optimization, purification and characterization of extracellular lipase produced by Serratia marcescens EGHK-19', Egyptian Journal of Botany, 64(4), pp. 107-119. doi: 10.21608/ejbo.2024.244359.2545
Issa, H., Abou-Dobara, M., El-Sayed, A., El-Bana, M. Optimization, purification and characterization of extracellular lipase produced by Serratia marcescens EGHK-19. Egyptian Journal of Botany, 2024; 64(4): 107-119. doi: 10.21608/ejbo.2024.244359.2545
Optimization, purification and characterization of extracellular lipase produced by Serratia marcescens EGHK-19
1Botany department, faculty of science, Port Said university, Port Said, Egypt
2Botany & Microbiology Department, Faculty of Science, Damietta University, Damietta El-Gededa, Egypt.
3Botany and Microbiology Department, Faculty of Science, Damietta University, Damietta El-Gededa, Egypt
4Department of Botany, Faculty of Science, Port Said University, Egypt
Abstract
Lipases are hydrolytic enzymes which have significant potential for commercial applications, particularly in the breakdown of oil contaminants. Serratia marcescens EGHK-19 isolate exhibited considerable lipase activity. This study investigates the optimization, purification, and characterization of lipase from the Serratia marcescens EGHK-19 isolate. The optimized culture conditions revealed that maximal lipase activity was achieved after 24 hours at 30°C and pH 7, with continuous shaking at 150 rpm. Utilizing a 2% inoculum percentage with 1% diesel and 0.3% tryptone in the presence of Fe2+, Ca2+, Mg2+ salts, and Tween 80 resulted in the highest activity at 17.278 U/ml/min. The purification process involved acetone precipitation and DEAE-Sephadex column chromatography, revealing a molecular weight of approximately 60 kDa on SDS-PAGE. This method exhibited a 0.985-fold purification and the final yield was limited to 2.097% due to lipase aggregates. Characterization of the purified lipase indicated optimal activity (8.765 U/mL/min) at 40°C and pH 7. The Km and Vmax values were calculated as 6.89 mM and 65.79 μmol/min, respectively. The presence of SDS, Tween 80, and Triton X-100 surfactants resulted in the inhibition of lipase activity. Despite these inhibitors, the biochemical characteristics of the purified lipase suggest its potential as an excellent candidate for various industrial applications.
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