Heterologous expression and N-terminal His-tagging processes affect the catalytic properties of staphylococcal lipases: A monolayer study

Horchani, Habib; Lignon, Sabrina; Lebrun, Régine; Sayari, Adel; Gargouri, Youssef; Verger, Robert

doi:10.1016/j.jcis.2010.07.021

Cited by 24 publications

(21 citation statements)

References 57 publications

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“…It is worth noting from Figs. 2 and 3 that the surface pressure thresholds of the staphylococcal lipases were found to range from 10 to 20 mN m À1 , which is in agreement with previous findings (9-14 mN m À1 ) obtained using the three dicaprin isomers as substrates [48]. These data suggest …”

Section: Surface Pressure Profiles Of Lipase Activitysupporting

confidence: 91%

Staphylococcal lipases stereoselectively hydrolyse the sn-2 position of monomolecular films of diglyceride analogs. Application to sn-2 hydrolysis of triolein

Horchani¹,

Salem²,

Châari³

et al. 2010

Journal of Colloid and Interface Science

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Section: Surface Pressure Profiles Of Lipase Activitysupporting

confidence: 91%

Staphylococcal lipases stereoselectively hydrolyse the sn-2 position of monomolecular films of diglyceride analogs. Application to sn-2 hydrolysis of triolein

Horchani¹,

Salem²,

Châari³

et al. 2010

Journal of Colloid and Interface Science

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“…Although this was reported to occur without any significant accu-mulation of partial glycerides, a different study demonstrated that S. aureus lipase SAL3 produced 1,3-diolein and lesser amounts of 1,2-or 2,3-diolein as the initial products of trioleoylglycerol hydrolysis, suggesting a preference for initial processing at the sn2 position. This lipase was designated SAL3, although it is evident from the primers that were employed for cloning of the gene (58,59) that it corresponds to SAL2 (SAUSA300_0320) described in this study. Nonetheless, both studies demonstrated hydrolysis of fatty acid esters at all three positions of the triacylglycerol substrate, suggesting that SAL2 can achieve complete conversion of triacylglycerol into glycerol and free fatty acids.…”

Section: Discussionmentioning

confidence: 99%

Role of Lipase from Community-Associated Methicillin-Resistant Staphylococcus aureus Strain USA300 in Hydrolyzing Triglycerides into Growth-Inhibitory Free Fatty Acids

et al. 2014

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Part of the human host innate immune response involves the secretion of bactericidal lipids on the skin and delivery of triglycerides into abscesses to control invading pathogens. Two Staphylococcus aureus lipases, named SAL1 and SAL2, were identified in the community-associated methicillin-resistant S. aureus strain USA300, which, presumably, are produced and function to degrade triglycerides to release free fatty acids. We show that the SAL2 lipase is one of the most abundant proteins secreted by USA300 and is proteolytically processed from the 72-kDa proSAL2 to the 44-kDa mature SAL2 by the metalloprotease aureolysin. We show that spent culture supernatants had lipase activity on both short-and long-chain fatty acid substrates and that deletion of gehB, encoding SAL2, resulted in the complete loss of these activities. With the use of gas chromatography-mass spectrometry, we show that SAL2 hydrolyzed trilinolein to linoleic acid, a fatty acid with known antistaphylococcal properties. When added to cultures of USA300, trilinolein and, to a lesser extent, triolein inhibited growth in a SAL2-dependent manner. This effect was shown to be due to the enzymatic activity of SAL2 on these triglycerides, since the catalytically inactive SAL2 Ser412Ala mutant was incapable of hydrolyzing the triglycerides or yielding delayed growth in their presence. Overall, these results reveal that SAL2 hydrolyzes triglycerides of both short-and long-chain fatty acids and that the released free fatty acids have the potential to cause significant delays in growth, depending on the chemical nature of the free fatty acid.

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“…In contrast, with a partly water soluble substrate (TC 4 ) this above mentioned negative effect is minimum [28,29]. In the light of the data obtained in previous study, it can be assumed that independently from the negative effects of the recombinant expression process per se, the presence of an N-terminal tag extension decrease the catalytic activities of staphylococcal lipases by creating a steric hindrance during the interfacial binding step [28,29]. An alternative hypothesis is that the N-terminal tag extension process may be responsible for a change in the orientation of the lipase at the interface.…”

Section: Expression and Purification Of The R-sxl2mentioning

confidence: 97%

Expression, purification of a novel alkaline Staphylococcus xylosus lipase acting at high temperature

Bouaziz¹,

Horchani²,

Salem³

et al. 2011

Biochemical Engineering Journal

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a b s t r a c tA new Staphylococcus xylosus strain was isolated. The extracellular lipase of S. xylosus (wt-SXL2) was purified to homogeneity from the culture medium. The specific activity of the purified enzyme, measured at pH 8.5 and 55• C using tributyrin or olive oil emulsion, reached, respectively, 6300 U/mg or 2850 U/mg. The sequenced 18 N-terminal amino acid showed a high degree of identity with known staphylococcal lipase sequences.The gene encoding the mature lipase was cloned and sequenced. The deduced amino acid sequence showed a significant similarity with various staphylococcal lipases. The highest overall identity (98.74%) was found with S. xylosus lipase (SXL1). The mature part of the lipase was expressed in Escherichia coli. The recombinant lipase was purified by affinity chromatography. The specific activity of the recombinant lipase was 4100 or 1500 U/mg using tributyrin or olive oil emulsion as substrate, respectively, at pH 8.5 and 55• C. The wild type and recombinant lipases presented a quite interesting thermal stability, after an incubation of 60 min at 55• C and they are found to be highly stable at a pH ranging from 4 to 11. Due to its stability at high temperature and in organic solvent, the wt-SXL2 was used as biocatalyst to synthesise a high added value molecules.

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Heterologous expression and N-terminal His-tagging processes affect the catalytic properties of staphylococcal lipases: A monolayer study

Cited by 24 publications

References 57 publications

Staphylococcal lipases stereoselectively hydrolyse the sn-2 position of monomolecular films of diglyceride analogs. Application to sn-2 hydrolysis of triolein

Staphylococcal lipases stereoselectively hydrolyse the sn-2 position of monomolecular films of diglyceride analogs. Application to sn-2 hydrolysis of triolein

Role of Lipase from Community-Associated Methicillin-Resistant Staphylococcus aureus Strain USA300 in Hydrolyzing Triglycerides into Growth-Inhibitory Free Fatty Acids

Expression, purification of a novel alkaline Staphylococcus xylosus lipase acting at high temperature

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