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

• Autorzy: Mondal K. , Malhotra S. P. , Jain V. , Singh R.
• Języki publikacji: EN

Abstrakty

EN

Employing the techniques of (NH₄)₂SO₄ 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 Km 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 Ki 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.

• Wydawca: -
• Czasopismo: Acta Physiologiae Plantarum
• Rocznik: 2009
• Tom: 31
• Numer: 1
• Opis fizyczny: p.81-87,fig.,ref.

Twórcy

autor

Mondal K.

• Plant Biochemistry and Molecular Biology Laboratory, Department of Biochemistry, CCS Haryana Agricultural University, Hisar 125 004, Haryana, India

autor

Malhotra S. P.

• Plant Biochemistry and Molecular Biology Laboratory, Department of Biochemistry, CCS Haryana Agricultural University, Hisar 125 004, Haryana, India

autor

Jain V.

• Plant Biochemistry and Molecular Biology Laboratory, Department of Biochemistry, CCS Haryana Agricultural University, Hisar 125 004, Haryana, India

autor

Singh R.

• Plant Biochemistry and Molecular Biology Laboratory, Department of Biochemistry, CCS Haryana Agricultural University, Hisar 125 004, Haryana, India

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