Purification, Biochemical and Kinetic Characterization of a Novel Alkaline sn-1,3-Regioselective Triacylglycerol Lipase from Penicilliumcrustosum Thom Strain P22 Isolated from Moroccan Olive Mill Wastewater

A novel extracellular lipase from a filamentous fungus Ascomycota strain, P22, was isolated from olive mill wastewater, then purified and characterized. This strain was identified as Penicillium crustosum Thom based on sequencing analyses. Penicilliumcrustosum Thom strain P22 lipase (PCrL) was purified 63-fold to homogeneity using ammonium sulfate precipitation and chromatography on a Q-Sepharose Fast Flow column, with a total yield of 34%. The purified PCrL had a molecular mass of 28 kDa, estimated by SDS-PAGE. The 20 NH2-terminal amino-acid residues showed a high degree of homology with those of other Penicillium lipases. The specific activity of PCrL at pH 9 and 37 °C were found to be 5000 and 10,000 U/mg on olive oil and trioctanoin emulsions, respectively. PCrL exhibited clear regioselectivity toward the sn-1 position of the surface-coated triglycerides which were esterified with α-eleostearic acid at the sn-1/3 position. PCrL was completely inhibited by 53 µM of Orlistat, 5 mM of phenylmethylsulfonylfluoride, and 2 mM of diiodopropyl fluorophosphate, suggesting that it belonged to the serine lipase family. PCrL showed high activity and stability in the presence of water-immiscible organic solvents, surfactant, and oxidizing agents, and showed considerable compatibility with commercial laundry detergents. Washing performance analysis revealed that it could effectively remove oil stains. Hence, PCrL has several attractive properties that make it a promising potential candidate for detergent formulations

IN VITRO BIODEGRADATION OF OLEUROPEIN BY LACTOBACILLUS PLANTARUM FSO175 IN STRESS CONDITIONS (pH, NaCl AND GLUCOSE)

International audienceThe objective of this work is to study the oleuropein (OLP) biodegradation by a strain of Lactobacillus plantarum FSO175 under conditions (pH 4.5, NaCl 5% and glucose 1%), during 7 days of incubation at 30 °C, in modified MRS broth containing OLP as carbon source. The results obtained, by HPLC analyses, showed that the biodegradation of OLP by L. plantarum FSO175, is accompanied with increase of hydroxytyrosol (HT) content and acidity values. The yields of OLP degradation and HT accumulation are depending on carbon sources, stress conditions and time of incubation. So that, the drastic reduction in OLP biodegradation, obtained at pH 6.7 (trial B), decreased significantly (p <0.05) in presence of NaCl 5% (trial D) and with glucose 1% (trial C). Indeed, the OLP biodegradation rate was maximal at pH 4.5 (trial E) and was higher even with NaCl 5% (trial F). In contrast, in presence of combined stress conditions (glucose 1%, pH 4.5 and NaCl 5%: trial G), the biodegradation of OLP decreased significantly (p <0.05). The effectiveness of the strain L. plantarum FSO175 in OLP biodegradation leading to variable yields of HT production, revealed its promising perspectives as starter culture, under controlled stress conditions of pH 4.5 and NaCl 5%, allowing the production of green table olives rich of HT, the main antioxidant highly desired in foods