Purification and biochemical characterization of pancreatic phospholipase A2 from the common stingray Dasyatis pastinaca

Abstract

BackgroundMammalian sPLA2-IB are well characterized. In contrast, much less is known about aquatic ones. The aquatic world contains a wide variety of living species and, hence represents a great potential for discovering new lipolytic enzymes.ResultsA marine stingray phospholipase A2 (SPLA2) was purified from delipidated pancreas. Purified SPLA2, which is not glycosylated protein, was found to be monomeric protein with a molecular mass of 14 kDa. A specific activity of 750 U/mg for purified SPLA2 was measured at optimal conditions (pH 8.5 and 40 °C) in the presence of 4 mM NaTDC and 8 mM CaCl2 using PC as substrate. The sequence of the first twenty first amino-acid residues at the N-terminal extremity of SPLA2 was determined and shows a close similarity with known mammal and bird pancreatic secreted phospholipases A2. SPLA2 stability in the presence of organic solvents, as well as in acidic and alkaline pH and at high temperature makes it a good candidate for its application in food industry.ConclusionsSPLA2 has several advantageous features for industrial applications. Stability of SPLA2 in the presence of organic solvents, and its tolerance to high temperatures, basic and acidic pH, makes it a good candidate for application in food industry to treat phospholipid-rich industrial effluents, or to synthesize useful chemical compounds.

Recombinant Lipases and Phospholipases and Their Use as Biocatalysts for Industrial Applications

“…Porcine PLA 2 is also suitable for enzymatic degumming of edible oils [ 75 ]. Recently, the starfish Asterina pectinifera [ 78 ] and the common stingray Dasyatis pastinaca [ 79 , 80 ] have been reported as potential new sources of PLA 2 with potential application as biocatalysts, thus providing alternatives to porcine pancreatic PLA 2 .…”

“…Indeed, after incubation in hydrophobic organic solvents, TLPLB retained 91% to 161% of its phospholipase activity, while in hydrophilic solvents its residual activity was only 10% to 38% [ 104 ]. Recently, the two PLA 2 s from pancreas [ 79 ] and intestine [ 80 ] of the common stingray were shown to retain 100% of their activity in incubations with polar organic solvents; increased activity was even observed in the presence of ethanol and acetonitrile (by 12% to 23%).…”

“…Both of these PLA 2 s show their optimal activity at 90 °C and retain most of their activity after incubation at temperatures ranging from 80 to 100 °C. As well as their stability in organic solvents, the two PLA 2 s from the common stingray [ 79 , 80 ] and the TLPLB [ 104 ] were shown to be also thermotolerant. Indeed, these PLA 2 s showed optimal activity at 40 to 45 °C and maintained 75% to 80% of their maximal activity after incubation at 70 °C; similarly, TLPLB showed its maximal activity at 70 °C, and maintained 50% of this activity at temperatures from 30 to 100 °C.…”

“…Among the phospholipases, stability at extreme temperatures is often associated with stability at extreme pHs. Indeed, the two thermostable PLA 2 s from common stingray [ 79 , 80 ], the thermostable PLD 528 from S. olivochromogenes [ 114 ], and the metagenome-derived cold-active MPlaG [ 129 ] have pH optima at 8.0–9.5 and have been shown to be stable in the alkaline range.…”

Efecto antiinflamatorio de apitoxina de Apis mellifera sobre prostaglandina E2 del fluido crevicular gingival de pacientes con y sin enfermedad periodontal, sometidos a apiterapia: ensayo preliminar

“…De acuerdo a Brochetto-Braga y cols (11) , el veneno de Apis mellifera está compuesto por péptidos como la melitina, apamina, MCD, péptido 401, entre otros; y enzimas como la fosfolipasa A 2 y hialuronidasa, causantes de la acción antiinflamatoria (12) . Actualmente no existe un consenso general sobre un responsable específico de la acción antiinflamatoria.…”