Purification and biochemical characterization of an organic solvent‐tolerant and detergent‐stable lipase from Staphylococcus capitis

Abstract

A mesophilic bacterial culture, producing an extracellular alkaline lipase, was isolated from the gas‐washing wastewaters generated from the Sfax phosphate plant of the Tunisian Chemical Group and identified as Staphylococcus capitis strain. The lipase, named S. capitis lipase (SCL), has been purified to homogeneity from the culture medium. The purified enzyme molecular weight was around 45 kDa. Specific activities about 3,900 and 500 U/mg were measured using tributyrin and olive oil emulsion as substrates, respectively at 37°C and pH 8.5. Interestingly, the SCL maintained more than 60% of its initial activity over a wide pH values ranging from 5 to 11 with a high stability between pH 9 and 11 after 1 hr of incubation at room temperature. The lipase activity was enhanced in the presence of 2 mM of Mg2+, Ca2+, and K+. SCL showed significant stability in the presence of detergents and organic solvents. Altogether, these features make the SCL useful for industrial applications. Besides, SCL was compatible with commercially available detergents, and its incorporation increases lipid degradation performances making it a potential candidate in detergent formulation.

Production, purification and biochemical characterisation of a novel lipase from a newly identified lipolytic bacterium Staphylococcus caprae NCU S6

“…Results are presented as means ± standard deviation (n ¼ 3). growth phase of NCU S6 started quickly after culturing in fermentation medium for 3 h and lasted for approximately 21 h. A slow growth from 24 to 39 h was observed, and the strain subsequently entered a stationary phase from 42 to 111 h. Figure 3 also showed the dynamic changes of lipase activity, with an increase from the beginning of inoculation to 51 h. The growth rate declined from 51 to 111 h, which may result from the depletion of nutrients in the fermentation medium and a low metabolism level under aging phase of growth 16 . Obviously, the peak (51 h) of lipase activity (101 U/mL) in the culture period was the optimal enzyme production time.…”

Microbial lipases and their industrial applications: a comprehensive review

“…Lipases displayed pH dependent activities, generally at neutral pH 7.0 or up to pH 4.0 and 8.0 lipases are stable, Chromobacterium viscosum, A. niger and Rhizophus sp., produced extracellular lipases are active at acidic pH, and P. nitroaeducens produced alkaline lipase and active at pH 11.0 [ 54 ]. Under certain experimental conditions lipases have capability to reversing the reactions which leads to esterification and interesterification in the absence of water [ 55 , 56 ]. For the expression of lipase activities the cofactors are not necessary but calcium is the divalent cation stimulates the activity [ 57 , 58 ].…”

Polypropylene as a selective support for the immobilization of lipolytic enzymes: hyper‐activation, purification and biotechnological applications

“…12 The purification of microbial lipases has been achieved by ammonium sulfate precipitation, heat or alkaline treatment, acetone precipitation, or ultrafiltration, followed by a combination of chromatographic methodologies, such as gel filtration, hydrophobic interaction, anion or cation exchange, and affinity chromatography, as well as other methodologies, such as aqueous two-phase systems, reversed micellar systems, and preparative isoelectrofocusing. 4,22,24,53,77,[109][110][111] Due to the interfacial activation of lipases, recovery of these enzymes has been mostly performed by hydrophobic interaction chromatography through the adsorption on hydrophobic matrices, such as alginate, Sepharose preparations (octyl-, phenyl-, polyethylene glycol-, polypropylene glycol-), methylated silica, glass beads coated with hydrophobic materials, and polypropylene particles. 4,[112][113][114][115][116][117] Purification of lipases by adsorption-desorption on polypropylene particles was first reported by Gupta et al, 77 who used Accurel® MP-1000 to purify an alkaline lipase from Burkholderia multivorans directly from fermentation broth with a 3-fold purification and a 66% yield.…”