title: Characterization of esterases from Cucurbita pepo cv. "Eskandrani".
abstract: Two of the six esterases identified in Cucurbita pepo cv. "Eskandrani" were purified to homogeneity using two chromatography steps: anion exchange and gel filtration. The molecular weights of C. pepo esterases EIc and EII were 50,000 +/- 1500 and 68,000 +/- 1900 Da from gel filtration and 47,000 and 66,000 Da from SDS/PAGE, respectively, suggesting a monomeric structure for both enzymes. Esterases EIc and EII had K(m) values of 1.22 and 1.56 mM and pH optima at 9.0 and 8.0, respectively. The substrate specificity of C. pepo esterases EIc and EII were determined for a number of p-nitrophenyl esters, where their affinity toward these substrates were decreased as carbon atom number increased. Esterases EIc and EII had the same temperature optima, 40 degrees C. Thermal stability studies of esterases EIc and EII indicated that half maximal activities of EIc and EII esterases were reached at 55 degrees C and 50 degrees C, while they lost 45%, 51% and 70%, 77% of their activities after 30 and 90 min of incubation at 40 degrees C, respectively. The effect of different metal cations and inhibitors were examined. The inhibition studies revealed that the active sites of the two esterases contain serine and cysteine residues. The characteristics of C. pepo esterases are closely similar to those of microbial esterases used in food processing and food industry.
authors.affiliation: Molecular Biology Department, National Research Centre, Cairo, Egypt.
authors.forename: Afaf S
authors.initials: AS
authors.lastname: Fahmy
authors.forename: Amal Z
authors.initials: AZ
authors.lastname: Abo-Zeid
authors.forename: Tarek M
authors.initials: TM
authors.lastname: Mohamed
authors.forename: Hala M
authors.initials: HM
authors.lastname: Ghanem
authors.forename: Ibrahim H
authors.initials: IH
authors.lastname: Borai
authors.forename: Saleh A
authors.initials: SA
authors.lastname: Mohamed
mesh_terms: D002845:Chromatography; D002850:Chromatography, Gel; D028464:Cucurbita / Q000201:enzymology*; D004950:Esterases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D021122:Protein Subunits / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D021122:Protein Subunits; D004950:Esterases