Purification and Characterization of Four Pectinesterases from Sweet Cherry (<i>Prunus avium</i>L.)

Alonso, Jesús; Rodríguez, María Teresa; Canet, Wenceslao

doi:10.1021/jf960204s

Cited by 20 publications

(20 citation statements)

References 42 publications

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“…The carrot PE enzyme was competitively inhibited by D ‐galacturonic acid (Fig 6), with a K i of approximately 1 m M . Inhibition by D ‐galacturonic acid has also been reported in cherries38 and Erwinia chrysanthemy 39. This inhibition may be relevant to the control of enzymatic activity in vivo and could be especially important for the control of degradation processes in carrot pectic material.…”

Section: Resultsmentioning

confidence: 85%

Purification and characterisation of carrot (Daucus carota L) pectinesterase

et al. 2003

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A pectinesterase isoform with an alkaline isoelectric point of over 8.66 was detected in crude extracts of carrot. The enzyme was purified by ion exchange and molecular exclusion chromatography. The molecular weight of the isoform was 25 kDa, determined in native conditions by filtration through Sephadex G-75 SF. The enzyme showed a high affinity for its substrate, with K m and V max values of 0.031 mg ml −1 and 6.77 units respectively for apple pectin. The pectinesterase activity exhibited an optimum around pH 7.4 and was activated by metallic ions, with optimum activities at NaCl concentrations between 130 and 330 mM and at CaCl 2 concentrations between 15 and 50 mM. The enzyme was activated most by Ca 2+ and exhibited a greater tolerance of high concentrations of Na + . Comparison of its heat stability with other pectinesterases of vegetable origin indicated that the purified isoform was very thermolabile, being rendered inactive by heating for 5 min at 70 • C. The enzyme was inhibited by high concentrations of polygalacturonic acid and competitively inhibited by D-galacturonic acid, with a K i value of 1 mM.

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Section: Resultsmentioning

confidence: 85%

Purification and characterisation of carrot (Daucus carota L) pectinesterase

et al. 2003

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“…However, at 90°C, the enzyme was completely inactive after 45 min of pre-incubation. Thermostable forms of the enzyme have been reported in persimmon (Alonso et al 1997), sweet cherry (Alonso et al 1996), lemon endocarp (MacDonald et al 1993, orange (Amaral et al 2005), plums (Nunes et al 2006) and bergamot fruit (Laratta et al 2008).…”

Section: Characterization Of Pmementioning

confidence: 99%

Partial purification and characterization of pectinmethylesterase from ripening guava (Psidium guajava L.) fruits

et al. 2008

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Employing the techniques of (NH 4 ) 2 SO 4 fractionation, ion exchange chromatography on DEAE cellulose and gel filtration through Sephadex G-100, pectinmethylesterase (EC 3.1.1.11) was purified from guava (Psidium guajava L.) fruits var. Hisar Safeda harvested at turning stage of maturity to 129-fold with 28% recovery. Molecular weight as determined by gel filtration was found to be 51 kDa and the enzyme preparation exhibited the same molecular weight under native (Native-PAGE) and denaturating conditions (SDS-PAGE) indicating that the enzyme was a monomer. With pectin as the substrate, it exhibited the Michaelis Menten kinetics with K m value of 3.1 g l -1 . The enzyme was found to be stimulated by Ca ?? and Na ? and inhibited competitively by Dgalacturonic acid with K i value of 1.97 mM. The enzyme was completely inactivated by iodine while with diethyl pyrocarbonate and N-acetylimidazole, the enzyme was inhibited up to the extent of 56 and 45%, respectively. However, DTNB had no inhibitory effect whatsoever precluding the participation of any -SH group in the active centre. It is tentatively proposed that the enzyme has tyrosine and histidine residues at its active centre.

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“…The thermal inactivation of pepper PME at pH 7.5 in this temperature range exhibits a biphasic model, indicating the presence of a heat-labile and a heat-resistant fraction of PME, both showing first-order inactivation mechanisms. Labile and resistant forms of PME have been shown to occur in a number of other fruits and vegetables including oranges (18,31,32), grapefruits (33), sweet cherries (8), persimmon (9), tomatoes (28), and green beans (34). At the higher temperatures, the inactivation of the heat-labile fraction proceeds very quickly so that the inactivation rate constants cannot be accurately estimated.…”

Section: Purification Of Pepper Pectin Methylesterase By Affinitymentioning

confidence: 99%

Activity and Process Stability of Purified Green Pepper (Capsicum annuum) Pectin Methylesterase

Castro

Loey

Saraiva

et al. 2004

J. Agric. Food Chem.

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Pectin methylesterase (PME) from green bell peppers (Capsicum annuum) was extracted and purified by affinity chromatography on a CNBr-Sepharose-PMEI column. A single protein peak with pectin methylesterase activity was observed. For the pepper PME, a biochemical characterization in terms of molar mass (MM), isoelectric points (pI), and kinetic parameters for activity and thermostability was performed. The optimum pH for PME activity at 22°C was 7.5, and its optimum temperature at neutral pH was between 52.5 and 55.0°C. The purified pepper PME required the presence of 0.13 M NaCl for optimum activity. Isothermal inactivation of purified pepper PME in 20 mM Tris buffer (pH 7.5) could be described by a fractional conversion model for lower temperatures (55-57°C) and a biphasic model for higher temperatures (58-70°C). The enzyme showed a stable behavior toward high-pressure/temperature treatments.

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Purification and Characterization of Four Pectinesterases from Sweet Cherry (Prunus aviumL.)

Cited by 20 publications

References 42 publications

Purification and characterisation of carrot (Daucus carota L) pectinesterase

Purification and characterisation of carrot (Daucus carota L) pectinesterase

Partial purification and characterization of pectinmethylesterase from ripening guava (Psidium guajava L.) fruits

Activity and Process Stability of Purified Green Pepper (Capsicum annuum) Pectin Methylesterase

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