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
{"title":"亲和单片低温凝胶柱用于BanLec纯化的创新方法","authors":"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","doi":"10.1007/s10924-025-03610-8","DOIUrl":null,"url":null,"abstract":"<div><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 (− 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<sup>−1</sup>), swelling capacity (17.7 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 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<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 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.</p><h3>Graphical Abstract</h3><div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":659,"journal":{"name":"Journal of Polymers and the Environment","volume":"33 7","pages":"3428 - 3442"},"PeriodicalIF":5.0000,"publicationDate":"2025-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Affinity Monolithic Cryogel Column for an Innovative Approach in BanLec Purification\",\"authors\":\"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\",\"doi\":\"10.1007/s10924-025-03610-8\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><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 (− 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<sup>−1</sup>), swelling capacity (17.7 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 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<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 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.</p><h3>Graphical Abstract</h3><div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>\",\"PeriodicalId\":659,\"journal\":{\"name\":\"Journal of Polymers and the Environment\",\"volume\":\"33 7\",\"pages\":\"3428 - 3442\"},\"PeriodicalIF\":5.0000,\"publicationDate\":\"2025-05-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Polymers and the Environment\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s10924-025-03610-8\",\"RegionNum\":3,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"ENGINEERING, ENVIRONMENTAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Polymers and the Environment","FirstCategoryId":"5","ListUrlMain":"https://link.springer.com/article/10.1007/s10924-025-03610-8","RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"ENGINEERING, ENVIRONMENTAL","Score":null,"Total":0}
Affinity Monolithic Cryogel Column for an Innovative Approach in BanLec Purification
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.
期刊介绍：
The Journal of Polymers and the Environment fills the need for an international forum in this diverse and rapidly expanding field. The journal serves a crucial role for the publication of information from a wide range of disciplines and is a central outlet for the publication of high-quality peer-reviewed original papers, review articles and short communications. The journal is intentionally interdisciplinary in regard to contributions and covers the following subjects - polymers, environmentally degradable polymers, and degradation pathways: biological, photochemical, oxidative and hydrolytic; new environmental materials: derived by chemical and biosynthetic routes; environmental blends and composites; developments in processing and reactive processing of environmental polymers; characterization of environmental materials: mechanical, physical, thermal, rheological, morphological, and others; recyclable polymers and plastics recycling environmental testing: in-laboratory simulations, outdoor exposures, and standardization of methodologies; environmental fate: end products and intermediates of biodegradation; microbiology and enzymology of polymer biodegradation; solid-waste management and public legislation specific to environmental polymers; and other related topics.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
