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

Abstract

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.

Enzymatic destabilization of chemical surfactant in wastewater—a potent ultrafiltration foulant: kinetic studies

“…2(i)). They were observed to be similar to lipases reported from other substrates such as slaughter house waste, blood tissue lipid, beef tallow, and cooked sunflower oil with a molecular weight of 94 [23], 92 [24], 56 [16], and 39 kDa [15], respectively. The two lipases with different molecular weights may be due to the presence of soap oil and castor oil in TVFL.…”

“…It was found that the lipase contained polar amino acids by 40% and non-polar amino acids by 60%. The amino acid composition of the lipase (Table 2), unlike the lipases reported by other researchers [15,16,23,24], suggests that it contained a higher percentage (57.14%) of aliphatic amino acids. The molecular weight of lipase and its amino acid composition depend on the substrate used for its production.…”

“…Lipase activity was quantified by titrimetric assay, utilizing an olive oil emulsion with remote modifications as followed by Ramani and Sekaran [16]. One activity unit of lipase was defined as the amount of enzyme that relinquished 1 μM of fatty acid per minute under assay conditions.…”

Biodegradation of waste greases and biochemical properties of a novel lipase from Pseudomonas synxantha PS1

“…The LipPS1 was stable at pH 6.0-10.0 compared with that of Pseudomonas aeruginosa SL-72 (pH 7.0-8.0) (Verma et al 2012), Pseudomonas fluorescens (pH 7.0) (Panizza et al 2013), Pseudomonas cepacia (pH 7.0) (Li et al 2015) Pseudomonas gessardii (pH 5.0) (Ramani et al 2010) and Pseudomonas stutzeri PS59 (pH 8.5) (Li et al 2014). The optimum temperature (40°C) of the recombinant lipase is similar to lipase from P. gessardii, P. fluorescens, P. fragi and P. mendoncina, which were found to be optimally active within 35-45°C (Ramani and Sekaran 2012).…”

“…5c). The lipase derived from P. gessardii, P. fluorescens, P. fragi and P. mendoncina were found to be optimally active within 35-45°C (Ramani and Sekaran 2012). Residual activities were determined with standard assay conditions.…”