Preparation of high-density Concanavalin A adsorbent and its use for rapid, high-yield purification of peroxidase from horseradish roots

Fraguas, Laura Franco; Batista‐Viera, Francisco; Carlsson, Jan

doi:10.1016/j.jchromb.2003.12.023

Cited by 28 publications

(9 citation statements)

References 5 publications

(8 reference statements)

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“…The amount of biologically active Con A immobilized to the nanoparticles was quantitatively determined to be 7.5%. The % biologically active Con A is comparable to literature [23].…”

Section: Hrp Enzymatic Assaysupporting

confidence: 85%

Polymer-coated magnetic nanoparticles for the efficient capture of Mycobacterium tuberculosis (Mtb)

Smit

Lutz

2020

SN Appl. Sci.

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Modified chitosan and modified poly(styrene-alt-maleic anhydride) (SMA) were synthesized and utilized to coat superparamagnetic magnetite nanoparticles (SPMNs) to act as potential Mycobacterium tuberculosis (Mtb) capturing substrates. Chitosan and SMA were modified to form quaternary derivatives which have the potential to interact with the Mtb cell wall. The nano-substrates were also surface functionalized with a carbohydrate-binding protein, namely Concanavalin A (Con A), which can bind to the Mtb cell wall. Chitosan coated SPMNs were synthesized by in situ co-precipitating Fe 2+ and Fe 3+ with chitosan followed by further modification. SMA coated SPMNs were synthesized by activating the iron oxide core with 3-aminopropyl(triethoxysilane) followed by further modification. The nano-substrates were evaluated for their affinity and thus capturing capabilities utilizing the mCherry fluorophore tagged bacillus Calmette-Guérin (BCG) strain of Mycobacterium bovis, a live attenuated Mtb-mimic. Quaternary SMA (SMI-qC 12 ) revealed the highest overall affinity for BCG-mCherry (through electrostatic and hydrophobic interactions) whereas Con A immobilized chitosan had the highest affinity for the chitosan coated nanoparticles. The SPMNs were coated with three different polymer loadings and a dilution study was performed to determine the limit of detection. The 0.9 g loaded SMI-qC 12 SPMNs revealed the highest affinity for BCG-mCherry as determined via fluorescence microscopy and transmission electron microscopy.

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“…The amount of biologically active Con A immobilized to the nanoparticles was quantitatively determined to be 7.5%. The % biologically active Con A is comparable to literature [23].…”

Section: Hrp Enzymatic Assaysupporting

confidence: 85%

Polymer-coated magnetic nanoparticles for the efficient capture of Mycobacterium tuberculosis (Mtb)

Smit

Lutz

2020

SN Appl. Sci.

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“…However, the use of Immobead 150P to be a base for Con-A immobilization first and then, consequently, the contact between Con-A and the glycosyl moiety of laccase has not so far been stated. Biotechnologically, Con-A has been applied as an affinity ligand for the immobilization and purification of glycoconjugates and glycoproteins [38,39].…”

Section: Introductionmentioning

confidence: 99%

Laccase-Oriented Immobilization Using Concanavalin A as an Approach for Efficient Glycoproteins Immobilization and Its Application to the Removal of Aqueous Phenolics

2022

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An expanding number of human activities are contributing to the rising levels of aromatic compounds, which pose a major threat to the ecosystem. However, readily available microbial enzymes might be used to remediate contaminated wastewater in an economical and environmentally benign manner. In this study, an efficient method of laccase-oriented immobilization on modified Immobead 150P was proposed. The oriented immobilization technique using aminated laccase exceeds in both protein loading onto the carrier (4.26 mg/g) and immobilization yield (93.57%) due to the availability of more active sites. The oriented aminated laccase preserves 100% and 95% of its original activity after six and ten cycles of operation, respectively. The thermal stability performance of the oriented enzyme was the best among both free and random immobilized forms, since it was able to conserve 79% and 44% of its initial activity after 6 h at 50 °C and 60 °C, respectively. The ideal pH of oriented immobilized laccase was altered from 3.0 to 4.0, and it was more stable than both free and random immobilized laccases at pH 7.0. Finally, the integration of the adsorption capacity of Immobead 150P and the biodegradation ability of laccase promises the efficient removal of aqueous phenolics. Oriented immobilized laccase may provide a significant new approach for wastewater treatment, according to these findings.

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“…Lectins have mainly been obtained from the seeds of leguminous plants but also from many other plant and animal tissues (Santos et al, 2005(Santos et al, , 2009Rameshwaram & Nadimpalli, 2008;Wong & Ng, 2006). They are valuable tools in biotechnological research (Fraguas et al, 2004) and biomedical applications (Kelly et al, 2005;Favacho et al, 2007). An important application of immobilised lectins reported in the literature (Aniulyte et al, 2006) is the isolation of biomolecules (Kelly et al, 2005) i.e.…”

Section: Introductionmentioning

confidence: 99%

“…An important application of immobilised lectins reported in the literature (Aniulyte et al, 2006) is the isolation of biomolecules (Kelly et al, 2005) i.e. plasma glycoproteins (lecithin-cholesterol acytransferase) (Lima *Corresponding author, e-mail: regina@deb.uminho.pt et al, 1997), polysaccharides (Franco-Fraguas et al, 2003), trypsin inhibitor (Paiva et al, 2003), and enzymes (peroxidases) (Fraguas et al, 2004). Studies in the biomedical field reported the isolation of lectins with mitogenic activity on human lymphocytes (Maciel et al, 2004), antifungal activity (Yan et al, 2005), and activity against the proliferation of human cancer cell lines (Kaur et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Interaction of Moringa oleifera seed lectin with humic acid

et al. 2011

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The aim of this work was to characterise the affinity of protein preparations from Moringa oleifera seeds, specifically extract (seeds homogenised with 0.15 M NaCl), fraction (extract precipitated with 390 mg mL−1 of ammonium sulphate) and cMoL (coagulant M. oleifera lectin) to bind humic acids using a haemagglutinating activity assay with rabbit erythrocytes and a radial diffusion assay in agarose gel. Specific haemagglutinating activity (SHA) decreased by 94 % for the extract and cMoL and by 50 % for the fraction in the presence of humic acid. Precipitation bands were observed in the diffusion gel. Both results suggested humic acid-cMoL binding. Carbohydrates, potassium, and calcium ions and pH affected the SHA of cMoL. As an example of application, cMoL was immobilised on a column packed with sepharose receiving 20 mg mL−1 of carbon humic acid solution, 30 mg of humic acid per gram of support was removed. This result suggested that protein preparations might be used in water treatment to remove humic acids.

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Preparation of high-density Concanavalin A adsorbent and its use for rapid, high-yield purification of peroxidase from horseradish roots

Cited by 28 publications

References 5 publications

Polymer-coated magnetic nanoparticles for the efficient capture of Mycobacterium tuberculosis (Mtb)

Polymer-coated magnetic nanoparticles for the efficient capture of Mycobacterium tuberculosis (Mtb)

Laccase-Oriented Immobilization Using Concanavalin A as an Approach for Efficient Glycoproteins Immobilization and Its Application to the Removal of Aqueous Phenolics

Interaction of Moringa oleifera seed lectin with humic acid

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