Optimization, purification and characterization of extracellular lipase produced by Serratia marcescens EGHK-19 | ||
| Egyptian Journal of Botany | ||
| Article 9, Volume 64, Issue 4, December 2024, Pages 107-119 PDF (1.78 M) | ||
| Document Type: Special Issue (Original Article) | ||
| DOI: 10.21608/ejbo.2024.244359.2545 | ||
| Authors | ||
| Heba Kamal Issa* 1; Mohammed Ismaeil Abou-Dobara2; Ahmed El-Sayed3; Magdy Ibrahim El-Bana4 | ||
| 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. | ||
| Keywords | ||
| Submerged fermentation; DEAE-Sephadex; SDS-PAGE; aggregates | ||
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