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<p>Banana lectin (BanLec) is a glycoprotein with the ability to bind reversibly to carbohydrates present on the cell surface. It has several biotechnological applications, including antiviral, antitumor, antibacterial and anticancer activities. This study aimed was to purify the BanLec present in the fruit of the plantain (Musa spp.) using a supermacroporous monolithic column functionalized with D-glucosamine and to characterize its biological properties. To this end, a cryogel based on acrylamide and bis-acrylamide was developed under freezing conditions (−&#xa0;12&#xa0;°C) and then functionalized with D-glucosamine (pAAm-Glucosamine). The matrix was characterized in terms of its physical, morphological and hydrodynamic structure, including analyses of porosity (92.4%, with pore diameters between 4&#xa0;µm and 9&#xa0;µm), degree of expansion (21.4 L.kg<sup>−1</sup>), swelling capacity (17.7&#xa0;kg.kg<sup>−1</sup>), Fourier transform infrared spectroscopy (FTIR), residence time distribution (DTR), apparent axial dispersion coefficient (Dax), height of equivalent theoretical plates (HETP) and flow permeability (1.74 × 10<sup>−12</sup> m<sup>2</sup>), as well as scanning electron microscopy (SEM). BanLec was purified from 100&#xa0;g of crude plantain extract using the ÄKTA Pure 25 L system in triplicate. The purified lectin was characterized by SDS-PAGE, HPLC and haemagglutination assays to assess its biological properties. The yield obtained was 0.053&#xa0;mg of protein per gram of fruit, with positive reactivity in the hemagglutination test (titer of 64 HU and specific activity of 69.94 HU mg.mL<sup>−1</sup>). BanLec showed thermal stability and resistance to pH variations between 3.0 and 7.0, as well as specificity for glucose, glucosamine and mannose in carbohydrate inhibition tests. Analysis by SDS-PAGE revealed a band with a molecular mass of approximately 14.4&#xa0;kDa, confirmed by the ÄKTA system and by HPLC, in which elution resulted in a single peak with a molecular mass of 20.48&#xa0;kDa. The results obtained confirm that the approach used for the extraction and purification of BanLec was highly efficient, preserving its biological activity as demonstrated by hemagglutination assays. These results not only highlight the effectiveness of the method used, but also reinforce the promising potential of BanLec for various biotechnological applications.</p> Graphical Abstract <p></p>

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Affinity Monolithic Cryogel Column for an Innovative Approach in BanLec Purification

  • 期刊论文

  • 发表日期：

    2025年5月30日

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  • Ivonéa Soares do Nascimento,
  • Charline Soares dos Santos Rolim,
  • Alexandre Araújo Pimentel,
  • Jennifer Renata Brasil dos Santos,
  • Yara Gomes de Souza,
  • Ted Possidônio dos Santos,
  • Jonathan Barbosa Santos,
  • Renata Cristina Ferreira Bonomo,
  • Leandro Soares Santos,
  • Rafael da Costa Ilhéu Fontan

摘要

Banana lectin (BanLec) is a glycoprotein with the ability to bind reversibly to carbohydrates present on the cell surface. It has several biotechnological applications, including antiviral, antitumor, antibacterial and anticancer activities. This study aimed was to purify the BanLec present in the fruit of the plantain (Musa spp.) using a supermacroporous monolithic column functionalized with D-glucosamine and to characterize its biological properties. To this end, a cryogel based on acrylamide and bis-acrylamide was developed under freezing conditions (− 12 °C) and then functionalized with D-glucosamine (pAAm-Glucosamine). The matrix was characterized in terms of its physical, morphological and hydrodynamic structure, including analyses of porosity (92.4%, with pore diameters between 4 µm and 9 µm), degree of expansion (21.4 L.kg−1), swelling capacity (17.7 kg.kg−1), Fourier transform infrared spectroscopy (FTIR), residence time distribution (DTR), apparent axial dispersion coefficient (Dax), height of equivalent theoretical plates (HETP) and flow permeability (1.74 × 10−12 m2), as well as scanning electron microscopy (SEM). BanLec was purified from 100 g of crude plantain extract using the ÄKTA Pure 25 L system in triplicate. The purified lectin was characterized by SDS-PAGE, HPLC and haemagglutination assays to assess its biological properties. The yield obtained was 0.053 mg of protein per gram of fruit, with positive reactivity in the hemagglutination test (titer of 64 HU and specific activity of 69.94 HU mg.mL−1). BanLec showed thermal stability and resistance to pH variations between 3.0 and 7.0, as well as specificity for glucose, glucosamine and mannose in carbohydrate inhibition tests. Analysis by SDS-PAGE revealed a band with a molecular mass of approximately 14.4 kDa, confirmed by the ÄKTA system and by HPLC, in which elution resulted in a single peak with a molecular mass of 20.48 kDa. The results obtained confirm that the approach used for the extraction and purification of BanLec was highly efficient, preserving its biological activity as demonstrated by hemagglutination assays. These results not only highlight the effectiveness of the method used, but also reinforce the promising potential of BanLec for various biotechnological applications.

Graphical Abstract