Production of lipase from Pseudomonas gessardii using blood tissue lipid and thereof for the hydrolysis of blood cholesterol and triglycerides and lysis of red blood cells.
The study demonstrates the production of lipase (LIP) from Pseudomonas gessardii using blood tissue lipid as the substrate for the hydrolysis of blood cholesterol and triglycerides. The lipase was purified with the specific activity of 828 U/mg protein and the molecular weight of 56 kDa. The maximum lipase activity was observed at the pH 7.0 and the temperature 37 °C. The amino acid composition of purified lipase was determined by HPLC. The mesoporous activated carbon (MAC) was used for the immobilization of lipase for the repeated use of the enzyme catalyst. The K (m) value of immobilized lipase (MAC-LIP) and the free lipase (LIP) was 0.182 and 1.96 mM, respectively. The V (max) value of MAC-LIP and LIP was 1.33 and 1.26 mM/min, respectively. The MAC and MAC-LIP were characterized by scanning electron microscopy (SEM). The hydrolysis study showed 78 and 100% hydrolysis of triglycerides and cholesterol, respectively, for LIP and 84 and 100% hydrolysis of triglycerides and cholesterol, respectively, for MAC-LIP at the reaction time of 1 h. The effect of lipase on cell wall lysis was carried out on the RBCs of blood plasma. Interestingly, 99.9% lysis of RBCs was observed within 2 h. SEM images and phase contrast microscopy confirmed the lysis of RBCs. This work provides a potential biocatalyst for the hydrolysis of blood cholesterol and triglycerides.
Environmental Technology Division, Council of Scientific and Industrial Research, Central Leather Research Institute, Chennai, 600 020, Tamil Nadu, India. kandasamyramani@gmail.com
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Ramani
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Sekaran
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D002606:Charcoal / Q000737:chemistry; D002784:Cholesterol / Q000097:blood / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000737:chemistry*; D004912:Erythrocytes / Q000737:chemistry*; D006041:Goats; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification; D011549:Pseudomonas / Q000201:enzymology*; D014280:Triglycerides / Q000097:blood / Q000737:chemistry*
D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D014280:Triglycerides; D002606:Charcoal; D002784:Cholesterol; D008049:Lipase