Cloning, expression, purification and physical and kinetic characterization of the phosphoenolpyruvate carboxylase from orange (Citrus sinensis osbeck var. Valencia) fruit juice sacs

Publication Overview
TitleCloning, expression, purification and physical and kinetic characterization of the phosphoenolpyruvate carboxylase from orange (Citrus sinensis osbeck var. Valencia) fruit juice sacs
AuthorsPerotti VE, Figueroa CM, Andreo CS, Iglesias AA, Podestá FE
TypeJournal Article
Journal NamePlant science
Volume179
Issue5
Year2010
Page(s)527-535
CitationPerotti VE, Figueroa CM, Andreo CS, Iglesias AA, Podestá FE. Cloning, expression, purification and physical and kinetic characterization of the phosphoenolpyruvate carboxylase from orange (Citrus sinensis osbeck var. Valencia) fruit juice sacs. Plant science. 2010; 179(5):527-535.

Abstract

Phosphoenolpyruvate (PEP) carboxylase (PEPCase) from orange fruit juice sacs has been cloned and heterogously expressed in high yield. The purified recombinant enzyme displays properties typical of plant PEPCase, including activation by sugar phosphates and inhibition by malate and citrate. Malate inhibition is weak in the physiological pH range, and the enzyme is also poorly affected by Glu and Asp, known inhibitors of C₃ plants PEPCases. However, it is strongly inhibited by citrate. Orange fruit PEPCase phosphorylation by mammalian protein kinase A decreased inhibition by malate. The enzyme presents an unusual high molecular mass in the absence of PEP, while in its presence it displays a more common tetrameric arrangement. The overall properties of the enzyme suggest that it is suited for organic acid synthesis and NADH reoxidation in the mature fruit. The present study provides the first analysis of a recombinant fruit PEPCase.
Features
This publication contains information about 8 features:
Feature NameUniquenameType
CsiPEPC-FoCsiPEPC-Fogenetic_marker
CsiPEPC-int1CsiPEPC-int1genetic_marker
CsiPEPC-int2CsiPEPC-int2genetic_marker
CsiPEPC-int3CsiPEPC-int3genetic_marker
CsiPEPC-int4CsiPEPC-int4genetic_marker
CsiPEPC-int5CsiPEPC-int5genetic_marker
CsiPEPC-int6CsiPEPC-int6genetic_marker
CsiPEPC-ReCsiPEPC-Regenetic_marker
Stocks
This publication contains information about 1 stocks:
Stock NameUniquenameType
ValenciaValenciaaccession
Properties
Additional details for this publication include:
Property NameValue
Publication TypeJournal Article
Publication Date2010
Published Location|||
Language Abbreng
Publication Model[electronic resource].
URLhttp://dx.doi.org/10.1016/j.plantsci.2010.08.003
KeywordsCitrus sinensis, oranges, phosphoenolpyruvate carboxylase, gene expression, enzyme activity, enzyme kinetics, orange juice, recombinant proteins, enzyme activation, enzyme inhibitors, malic acid, citric acid, sugar phosphates, pH, protein phosphorylation, protein kinases, molecular weight, protein structure, orange juice sacs