Cibacron Blue F3GA ile modifiye polimerik mikroküreler ile RuBisCO adsorpsiyonu

Abstract

Bu çalışmada, Cibacron Blue F3GA (CB) ile modifiye edilmiş poli(hidrokisetil metakrilat) [PHEMA] mikroküreler hazırlandı ve ribuloz-1,5-bisfosfat karboksilaz/oksijenaz (RuBisCO) adsorpsiyonunda kullanıldı. PHEMA mikroküreler süspansiyon polimerizasyonu tekniği ile sentezlendi ve taramalı elektron mikroskopisi (SEM) ve Fourier transform infrared spektroskopisi (FTIR) ile karakterize edildi. CB ile kovalent olarak modifiye edilen CB-PHEMA mikroküreler RuBisCO adsorpsiyonunda kullanıldı. Adsorpsiyona CB içeriği, başlangıç RuBisCO derişimi (0,2-1.0 mg/mL), pH (5,5-9.0), sıcaklık (4°C, 22 oC ve 35°C) ve temas süresinin etkisi araştırıldı. CB-PHEMA mikrokürelerin CB içeriği elementel analiz ile belirlenen %N miktarı kullanılarak hesaplandı. CB-PHEMA mikrokürelere adsorplanan maksimum RuBisCO miktarı 33,59 mg/g olarak belirlendi (22 oC, pH 6,0). Üç farklı sıcaklıkta (4°C, 22 oC ve 35°C) elde edilen RuBisCO adsorpsiyon verilerinin izoterm ve kinetik modellere uygunluğu araştırıldı. RuBisCO adsorpsiyon verilerinin Freundlich izoterm modeli ve yalancı-ikinci derece kinetik modele uygun olduğu belirlendi. RuBisCO adsorpsiyonuna ilişkin Gibbs serbest enerji değişimi (ΔG ͦ) değerleri; 4 oC, 22 oC ve 35 oC sıcaklık için sırasıyla -9,22 kj/mol, -10,44 kJ/mol ve -11,33 kJ/mol olarak hesaplandı. Entalpi değişimi (ΔH ͦ) +9,661 kJ/mol ve entropi değişimi (ΔS ͦ) +68,14 J/mol.K olarak belirlendi. Termodinamik parametreler, CB-PHEMA mikrokürelere RuBisCO adsorpsiyonunun endotermik ve kendiliğinden gerçekleşen bir proses olduğunu kanıtladı. CB-PHEMA mikrokürelerin ıspanak protein ekstraktından RuBisCO adsorpsiyon performansı SDS-PAGE analizi ile değerlendirildi.

Keywords

• RuBisCO
• adsorpsiyon
• Cibacron Blue F3GA
• bitki proteom analizi

RuBisCO adsorption via polymeric microbeads modified with Cibacron Blue F3GA

Abstract

In this study, poly(hydroxyethyl methacrylate) [PHEMA] microbeads modified with Cibacron Blue F3GA (CB) were prepared and used for the adsorption of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO). PHEMA microbeads were synthesized by suspension polymerization and characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Then, CB modified CB-PHEMA microbeads were used for RuBisCO adsorption. Effects of CB content, initial RuBisCO concentration (0.2-1.0 mg/mL), pH (5.5-9.0), temperature (4 oC, 22 oC and 35°C) and contact time were investigated. The CB amount of the CB-PHEMA microbeads was calculated via N% determined by elemental analysis. The maximum RuBisCO amount adsorbed onto the CB-PHEMA microbeads was determined as 33.59 mg/g (22 oC, pH 6.0). The RuBisCO adsorption data obtained at three different temperatures (4°C, 22 oC and 35°C) were applied to isotherm and kinetic models. Freudlich isotherm model and pseudo-second order kinetic model were well fitted to adsorption data with high correlation coefficients. Gibss free energy changes (ΔG ͦ) of RuBisCO adsorption for 4 oC, 22 oC and 35 oC were calculated as -9.22 kj/mol, -10.44 kJ/mol and -11.33 kJ/mol, respectively. Enthalpy change (ΔH ͦ) was +9.661 kJ/mol and entrophy change (ΔS ͦ) was +68.14 J/mol.K. The calculated thermodynamic parameters showed that RuBisCO adsorption onto CB-PHEMA microbeads was endothermic and spontaneous. The RuBisCO adsorption performance of CB-PHEMA microbeads from spinach protein extract was evaluated by SDS-PAGE analysis.

Keywords

• RuBisCO
• adsorption
• Cibacron Blue F3GA
• plant proteomics

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