Substrate specificity and kinetic properties of enzymes belonging to the hormone-sensitive lipase family: Comparison with non-lipolytic and lipolytic carboxylesterases

Tetrahydrolipstatin (THL) is bactericidal but its precise target spectrum is poorly characterized. Here, we used a THL analog and activity-based protein profiling to identify target proteins after enrichment from whole cell lysates of Mycobacterium bovis Bacillus Calmette-Gué rin cultured under replicating and non-replicating conditions. THL targets ␣/␤-hydrolases, including many lipid esterases (LipD, G, H, I, M, N, O, V, W, and TesA). Target protein concentrations and total esterase activity correlated inversely with cellular triacylglycerol upon entry into and exit from non-replicating conditions. Cellular overexpression of lipH and tesA led to decreased THL susceptibility thus providing functional validation. Our results define the target spectrum of THL in a biological species with particularly diverse lipid metabolic pathways. We furthermore derive a conceptual approach that demonstrates the use of such THL probes for the characterization of substrate recognition by lipases and related enzymes. Molecular & Cellular

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“…4D); this is consistent with the biochemical results with synthetic substrates (Fig. 3E, 3F) (39,46). This approach therefore provides FIG.…”

Section: Figsupporting

confidence: 79%

Targeting Lipid Esterases in Mycobacteria Grown Under Different Physiological Conditions Using Activity-based Profiling with Tetrahydrolipstatin (THL)

Ravindran

Rao

Cheng

et al. 2014

Molecular & Cellular Proteomics

104

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“…The apparent K m and V max values of Lip501r for pNP-butyrate in the presence of 5 mM CaCl 2 were 1.8 ± 0.4 mM and 220 ± 20 units/mg (k cat = 160 s -1 as calculated from the theoretical molecular mass), respectively. The kinetic parameters of Lip501r were comparable with those of other bacterial lipases for pNP-butyrate (Chahinian et al, 2005).…”

Section: Characterization Of the Recombinant Enzymesupporting

confidence: 58%

Isolation and characterization of a thermostable lipase from <i>Bacillus thermoamylovorans</i> NB501

Yamada

Sawano

Iwase

et al. 2016

J. Gen. Appl. Microbiol.

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“…Most works are related to microbial lipases where one can verify that the specificity of the enzyme to the substrate varies as a function of the source and production process (fermentation and/or extraction). Tributyrin is a shortchain triglyceride and is easily hydrolyzable by lipases and esterases [20,21]. Lipase from Pseudomonas fluorescens HU380 showed higher activity in tributyrin (C4:0) [22], the same behavior observed for crude enzymatic extract from Colletotrichum gloesporioides [23].…”

Section: Hydrolysis Substratesmentioning

confidence: 59%

Studies on Immobilization and Partial Characterization of Lipases from Wheat Seeds (Triticum aestivum)

Pierozan

Oestreicher

Oliveira

et al. 2011

Appl Biochem Biotechnol

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The objective of this study was to provide some features on immobilization and partial characterization of lipases from wheat seeds. The optimum pH and temperature were found to be 5.5 and 32-37 °C, respectively. The stability of the concentrated enzymatic extract to high temperatures (25, 35, 45, and 55 °C) showed that the incubation of the extract at 55 °C led to its complete inactivation. The concentrated enzymatic extract kept 90% of its hydrolytic and esterification activities until 70 and 40 days of storage at 4 °C, respectively. The extract presented higher hydrolytic specificity to substrates of medium and long chains and higher esterification affinity to fatty acids of short and medium chains and alcohols with two and three carbon atoms. After the immobilization process using activated coal and sodium alginate as supports, an enhancement of about threefold in lipase activity was observed. The development of the present work permitted us to point out some characteristics of lipases from wheat seeds necessary for the proposition of new future industrial applications for this important biocatalyst.

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Substrate specificity and kinetic properties of enzymes belonging to the hormone-sensitive lipase family: Comparison with non-lipolytic and lipolytic carboxylesterases

Cited by 50 publications

References 30 publications

Targeting Lipid Esterases in Mycobacteria Grown Under Different Physiological Conditions Using Activity-based Profiling with Tetrahydrolipstatin (THL)

Targeting Lipid Esterases in Mycobacteria Grown Under Different Physiological Conditions Using Activity-based Profiling with Tetrahydrolipstatin (THL)

Isolation and characterization of a thermostable lipase from <i>Bacillus thermoamylovorans</i> NB501

Studies on Immobilization and Partial Characterization of Lipases from Wheat Seeds (Triticum aestivum)

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