Optimization, Purification and Characterization of Extracellular Lipase Produced by Serratia marcescens EGHK-19 | ||||
| Egyptian Journal of Botany | ||||
| Articles in Press, Accepted Manuscript, Available Online from 14 January 2024 PDF (1.58 MB) | ||||
| Document Type: Special Issue (Original Article) | ||||
| DOI: 10.21608/ejbo.2024.244359.2545 | ||||
 View on SCiNiTO | ||||
| Authors | ||||
Heba K. Issa  1; Mohamed I. Abou Dobara2; Ahmed K.A. El-Sayed2; Magdy I. El-Bana1
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| 1Department of Botany, Faculty of Science, Port Said University, Port Said, Egypt | ||||
| 2Department of Botany & Microbiology, Faculty of Science, Damietta University, Damietta ElGededa, 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. | ||||
| Keywords | ||||
| Aggregates; DEAE Sephadex; SDS-PAGE; Submerged fermentation | ||||
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