Purification and properties of an esterase from Cucurbita maxima fruit tissue

Nourse, Amanda; Schabort, J.C.; Dirr, Heini W.; Dubery, Ian A.

doi:10.1016/0031-9422(89)80016-0

Cited by 12 publications

(11 citation statements)

References 29 publications

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“…These results recall those obtained for esterases from Cucurbita pepo whose activity on p ‐nitrophenyl esters was strongly inhibited by the presence of Cu 2+ ions (Nourse et al . ). Mg 2+ ions had no significant effect on GmDE activity (Table ).…”

Section: Resultsmentioning

confidence: 97%

A High Salt-Tolerant Thermoactive Esterase from Golden Grey Mullet: Purification, Characterization and Kinetic Properties

Smichi¹,

Fendri²,

Gargouri³

et al. 2015

Journal of Food Biochemistry

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An esterase was purified from the golden grey mullet viscera using successively a Sephacryl S-100 gel filtration, an anion-exchange chromatography and a highperformance liquid chromatography filtration column. The pure esterase (GmDE) is a monomer that has a molecular mass of about 55 kDa, as determined by SDS-PAGE analysis. The purified enzyme displayed a specific activity of 100 U/mg on short-chain triacylglycerols at a temperature of 50C. GmDE is therefore a thermoactive enzyme as compared to other fish lipolytic enzymes that have been studied so far. No significant lipolytic activity was noticed when long-chain triacylglycerol (olive oil) was used as a substrate. It is worth noting that the pure esterase was active in the presence of salt concentrations as high as 0.8 M. The GmDE N-terminal amino acid sequence showed no similarities with that of other known fish esterases. Altogether, these results suggest that the GmDE is a member of a new group of digestive esterases belonging to vertebrates. PRACTICAL APPLICATIONSCharacterization of an esterase from low-value fish viscera and the use of digestive enzyme may add value to this discarded species. Furthermore, the activity and stability at alkaline pH may also find use in laundry detergents. The thermoactivity of the purified esterase makes it a good candidate for potential application in food processing operations. Finally, the stability of the enzyme in high salt concentrations suggests that it can be used as an additive in different processes (food, cosmetic and pharmaceutical operations).

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Section: Resultsmentioning

confidence: 97%

A High Salt-Tolerant Thermoactive Esterase from Golden Grey Mullet: Purification, Characterization and Kinetic Properties

Smichi¹,

Fendri²,

Gargouri³

et al. 2015

Journal of Food Biochemistry

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“…The purified kidney bean esterase was resolved as a single band, indicating an increase in purity after the initial purification. It is reported that most of the plant esterases have low molecular weights and contain a single polypeptide chain, with molecular weights in the range of 20–60 kDa …”

Section: Resultsmentioning

confidence: 99%

“…It is reported that most of the plant esterases have low molecular weights and contain a single polypeptide chain, with molecular weights in the range of 20-60 kDa. 24,[28][29][30] . From the SDS-PAGE, the molecular mass of kidney bean esterase is approximately 26.5 kDa.…”

Section: Resultsmentioning

confidence: 99%

Identification and characterization of a novel carboxylesterase from Phaseolus vulgaris for detection of organophosphate and carbamates pesticides

et al. 2018

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“…They have been purified from various plant sources including finger millet (Eleusine coracana) (Upadhya et al, 1985), Cucurbita maxima fruit tissue (Nourse et al, 1989), Jatropa curcas L. seeds (Staubmann et al, 1999), Avena fatua (Mohamed et al, 2000), tomato (Stuhlfelder et al, 2002) and Cucurbita pepo (Afaf S Fahmy et al, 2008) by employing different purification processes including ammonium sulphate fractionation, ion exchange chromatography and gel filtration chromatography. However, not all preparations have been shown to be homogeneous.…”

Section: Purificationmentioning

confidence: 99%

“…Many researchers reported the homogeneity of their preparations using PAGE (Nourse et al, 1989). Carboxylesterases have been purified to homogeneity and purity was established by both gel electrophoresis and isoelectric focussing (IEF) (Upadhya et al, 1985).…”

Section: Criteria Of Puritymentioning

confidence: 99%

Carboxylesterases from the seeds of an underutilized legume, Mucuna pruriens; isolation, purification and characterization

2011

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a b s t r a c tTwo carboxylesterases (ME-III and ME-IV) have been purified to apparent homogeneity from the seeds of Mucuna pruriens employing ammonium sulfate fractionation, cation exchange chromatography on CMcellulose, gel-permeation chromatography on Sephadex G-100 and preparative PAGE. The homogeneity of the purified preparations was confirmed by polyacrylamide gel electrophoresis (PAGE), gel-electrofocussing and SDS-PAGE. The molecular weights determined by gel-permeation chromatography on Sephadex G-200 were 20.89 kDa (ME-III) and 31.62 kDa (ME-IV). The molecular weights determined by SDS-PAGE both in the presence and absence of 2-mercaptoethanol were 21 kDa (ME-III) and 30.2 kDa (ME-IV) respectively, suggesting a monomeric structure for both the enzymes. The enzymes were found to have Stokes radius of 2.4 nm (ME-III) and 2.7 nm (ME-IV). The isoelectric pH values of the enzymes, ME-III and ME-IV, were 6.8 and 7.4, respectively. ME-III and ME-IV were classified as carboxylesterases employing PAGE in conjunction with substrate and inhibitor specificity. The K m of ME-III and ME-IV with 1-naphthyl acetate as substrate was 0.1 and 0.166 mM while with 1-naphthyl propionate as substrate the K m was 0.052 and 0.0454 mM, respectively. As the carbon chain length of the acyl group increased, the affinity of the substrate to the enzyme increased indicating hydrophobic nature of the acyl group binding site. The enzymes exhibited an optimum temperature of 45°C (ME-III) and 37°C (ME-IV), an optimum pH of 7.0 (ME-III) and 7.5 (ME-IV) and both the enzymes (ME-III and ME-IV) were stable up to 120 min at 35°C. Both the enzymes were inhibited by organophosphates (dichlorvos and phosphamidon), but resistant towards carbamates (carbaryl and eserine sulfate) and sulphydryl inhibitors (p-chloromercuricbenzoate, PCMB).

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Purification and properties of an esterase from Cucurbita maxima fruit tissue

Cited by 12 publications

References 29 publications

A High Salt-Tolerant Thermoactive Esterase from Golden Grey Mullet: Purification, Characterization and Kinetic Properties

A High Salt-Tolerant Thermoactive Esterase from Golden Grey Mullet: Purification, Characterization and Kinetic Properties

Identification and characterization of a novel carboxylesterase from Phaseolus vulgaris for detection of organophosphate and carbamates pesticides

Carboxylesterases from the seeds of an underutilized legume, Mucuna pruriens; isolation, purification and characterization

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