Purification and properties of extracellular lipase from Pseudomonas aeruginosa EF2

Abstract

Extracellular lipase was purified from a Tween 80-limited continuous culture of Pseudomonas aeruginosa EF2 by ultrafiltration of the culture supernatant followed by anion-exchange and gel-filtration FPLC. The lipase was composed of a single subunit (M, 29000, PI 4.9), which was capable of a variable degree of aggregation, and which exhibited both lipase activity, measured with the insoluble substrate olive oil (predominantly triolein), and esterase activity, measured with the soluble substrates p-nitrophenyl acetate and Tween 80. Lipase activity was approximately eight times higher than either type of esterase activity (kcat approximately 3OoOs-' for the hydrolysis of olive oil). The enzyme showed a marked regiospecificity for the 1,3-oleyl residues of radiolabelled triolein, was relatively stable at moderate temperatures (exhibiting a biphasic loss of activity with an initial fi of 17.5 min at 60 "C) and was very stable to freezing and thawing. Lipase activity was only weakly inhibited by the serine-active reagent 3,4-dichloroisocoumarin, and was not inhibited by the chelating agent EDTA (1 mM). The Nterminal amino acid sequence of the Ps. aeruginosa EF2 lipase showed a marked similarity to those of several other bacterial lipases.

Molecular genetics of the extracellular lipase of Pseudomonas aeruginosa PAO1

“…aeruginosa PAClR (Stuer et al, 1986;Jaeger et al, 1991Jaeger et al, , 1992 and of P . aeruginosa EF2 (Gilbert et al, 1991) yielded results which correspond very well to data deduced from the nucleotide sequence of the lipase gene of P. aeruginosa PAO1. These data comprise the molecular mass of the enzyme, the isoelectric point (calculated : PI = 5.6 ; analysed by isoelectric focussing : PI = 5.8), the overall amino acid composition and the amino acid sequence of the N-terminus of the secreted protein.…”

Production and Partial Characterization of Lipase from Pseudomonas putida

“…The results of optimum working temperature for lipase are confirmed by studies done by Gilbert et al [25] showed that lipase activity decreased after 45ºC while Zaliha et al [26] found lipase activity to be optimum above 40ºC. The results for varying optimum pH coincide with various studies carried out by Mubarak-Qamsari et al [21] which reveal similar results that the enzyme has the ability to work at pH 9 and 10.…”

Characterization and optimization of lipase activity produced by Pseudomonas monteilli 2403-KY120354 isolated from ground beef

“…KE38 was grown at optimum at 25°C, but its lipase had maximum lipase activity at 45°C. In contrast to our results, Gilbert et al (1991) indicated that maximum lipase activity by Pseudomonas spp. was detected at 30°C, while the results reported by Qamsari et al (2011) showed that maximum lipase activity produced by P. aeruginosa KM110 was at temperature range (35 to 45°C).…”