How do lipases and esterases work: the electrostatic contribution

Neves‐Petersen, Maria Teresa; Fojan, Peter; Petersen, Steffen B.

doi:10.1016/s0168-1656(00)00360-6

Cited by 125 publications

(65 citation statements)

References 31 publications

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“…3A), an unexpectedly alkaline pH for carboxylesterase. Protein electrostatics investigation suggested that the active site of esterases and lipases displayed a negative potential in the pH range associated with their maximum activity, 39) and that the esterases showed their optimum charge at pH 6 to 7, which correlated with their usually lower pHactivity optimum than that of the lipases around pH 8. 1,6) These investigations were carried out using esterases and lipases of which the 3D-structures and amino acid sequences had been experimentally determined.…”

Section: Characterization Of the Enzymementioning

confidence: 99%

A Novel Alkaline Esterase fromSporosarcinasp. nov. Strain eSP04 Catalyzing the Hydrolysis of a Wide Variety of Aryl-carboxylic Acid Esters

Takehara

Kinoshita

Miyamoto

et al. 2012

Bioscience, Biotechnology, and Biochemistry

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A novel esterase showing activity specific for esters of aryl-carboxylic acids was discovered in Sporosarcina sp. nov., which was identified by the 16S rDNA sequencing method in addition to morphological and physiological analyses. The aryl-carboxylesterase (named EstAC) was purified 780-fold from crude cell extracts by a 5-step procedure. EstAC was characterized as a monomeric protein with a molecular weight of 43,000, an optimum pH of around 9.0, and an optimum temperature of 40 C. The pH optimum and the effects of inhibitors together with an internal amino acid sequence suggested that EstAC is a member of family VIII esterases. EstAC was found to be highly active on a wide variety of substrates such as alkyl benzoates, alkyl phenylacetates, ethyl -or -substituted phenylpropionates, dialkyl terephthalates, dimethyl isophthalate, and ethylene glycol dibenzoate. However, monomethyl terephthalate was not hydrolyzed. It was suggested that EstAC had 4-hydroxybenzoyl and cinnamoyl esterase activities as well.

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Section: Characterization Of the Enzymementioning

confidence: 99%

A Novel Alkaline Esterase fromSporosarcinasp. nov. Strain eSP04 Catalyzing the Hydrolysis of a Wide Variety of Aryl-carboxylic Acid Esters

Takehara

Kinoshita

Miyamoto

et al. 2012

Bioscience, Biotechnology, and Biochemistry

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“…VolA demonstrated a partiality for alkaline pH. It is possible that at an alkaline pH, the active-site residues of VolA exist in a negatively charged state, allowing efficient release of the LCFA reaction product through electrostatic repulsion (35).…”

Section: Fig 7 Expression Of Aeromonas Hydrophilamentioning

confidence: 99%

Characterization of the Vibrio cholerae VolA Surface-Exposed Lipoprotein Lysophospholipase

2014

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bBacterial lipases play important roles in bacterial metabolism and environmental response. Our laboratory recently discovered that a novel lipoprotein lysophospholipase, VolA, localizes on the surface of the Gram-negative aquatic pathogen Vibrio cholerae. VolA functions to cleave exogenous lysophosphatidylcholine, freeing the fatty acid moiety for use by V. cholerae. This fatty acid is transported into the cell and can be used as a nutrient and, more importantly, as a way to alter the membrane architecture via incorporation into the phospholipid biosynthesis pathway. There are few examples of Gram-negative, surface-exposed lipoproteins, and VolA is unique, as it has a previously undercharacterized function in V. cholerae membrane remodeling. Herein, we report the biochemical characterization of VolA. We show that VolA is a canonical lipoprotein via mass spectrometry analysis and demonstrate the in vitro activity of VolA under a variety of conditions. Additionally, we show that VolA contains a conserved Gly-Xaa-Ser-Xaa-Gly motif typical of lipases. Interestingly, we report the observation of VolA homologs in other aquatic pathogens. An Aeromonas hydrophila VolA homolog complements a V. cholerae VolA mutant in growth on lysophosphatidylcholine as the sole carbon source and in enzymatic assays. These results support the idea that the lipase activity of surface-exposed VolA likely contributes to the success of V. cholerae, improving the overall adaptation and survival of the organism in different environments. R ecent work from our laboratory revealed the presence of a unique membrane-anchored lipase, VolA, localized on the surface of Vibrio cholerae cells (1). We demonstrated that VolA is required for growth when lysophosphatidylcholine (LPC) (Fig. 1) serves as the sole carbon source. We hypothesized that VolA could cleave exogenous lysophosphatidylcholine into long-chain fatty acid (LCFA) derivatives (Fig. 1), allowing them to be brought into the cell via a coexpressed fatty acid transporter, FadL. These data, taken together with the presence of a conserved lipase domain identified in V. cholerae (2) and the inability of wild-type V. cholerae to grow on phosphatidylcholine as a sole carbon source, suggest that VolA acts as a lysophospholipase in vivo.Phospholipases are members of the acylhydrolase family of enzymes, acting on ester bonds in phospholipid targets (3). The structure and function of such acylhydrolases are conserved and have been well studied. All contain a characteristic fold (4, 5), sharing a conserved lipase motif (2). In our previous work (1), we noted that VolA contains this conserved motif, which strongly implicates it as a lipase. Phospholipases vary in terms of their site of action on the phospholipid. Some target phosphate bonds, while others hydrolyze bonds between the glycerol moiety and the acyl chains. Most microbial lysophospholipases hydrolyze the ester bond between the acyl chain and the polar head group, releasing a free fatty acid (6). It is likely that VolA functions similarly to e...

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“…The optimum pH for most of the lipases is in the range 8.0-9.0 with few exceptions wherein the optimum pH lies between 5.0 and 6.0 (Neves-Petersen et al 2001).…”

Section: I E T H Y L E T H E Rmentioning

confidence: 99%

Kinetic modeling, production and characterization of an acidic lipase produced by Enterococcus durans NCIM5427 from fish waste

et al. 2013

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Enterococcus durans NCIM5427 (ED-27), capable of producing an intracellular acid stable lipase, was isolated from fish processing waste. Its growth and subsequent lipase production was optimized by Box Behneken design (optimized conditions: 5 % v/v fish waste oil (FWO), 0.10 mg/ml fish waste protein hydrolysates (FWPH) at 48 h of fermentation time). Under optimized conditions, ED-27 showed a 3.0 fold increase (207.6 U/ml to 612.53 U/ml) in lipase production, as compared to un-optimized conditions. Cell growth and lipase production was modeled using Logistic and LuedekingPiret model, respectively; and lipase production by ED-27 was found to be growth-associated. Lipase produced by ED-27 showed stability at low pH ranges from 2 to 5 with its optimal activity at 30°C , pH 4.6; showed metal ion dependent activity wherein its catalytic activity was activated by barium, sodium, lithium and potassium (10 mM); reduced by calcium and magnesium (10 mM). However, iron and mercury (5 mM) completely inactivated the enzyme. In addition, modifying agents like SDS, DTT, β-ME (1%v/v) increased activity of lipase of ED-27; while, PMSF, DEPC and ascorbic acid resulted in a marked decrease. ED-27 had maximum cell growth of 9.90309 log CFU/ml under optimized conditions as compared to 13 log CFU/ml in MRS. The lipase produced has potential application in poultry and slaughterhouse waste management.

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How do lipases and esterases work: the electrostatic contribution

Cited by 125 publications

References 31 publications

A Novel Alkaline Esterase fromSporosarcinasp. nov. Strain eSP04 Catalyzing the Hydrolysis of a Wide Variety of Aryl-carboxylic Acid Esters

A Novel Alkaline Esterase fromSporosarcinasp. nov. Strain eSP04 Catalyzing the Hydrolysis of a Wide Variety of Aryl-carboxylic Acid Esters

Characterization of the Vibrio cholerae VolA Surface-Exposed Lipoprotein Lysophospholipase

Kinetic modeling, production and characterization of an acidic lipase produced by Enterococcus durans NCIM5427 from fish waste

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