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Amino acid-based hydrophobic affinity cryogel for protein purification from ora-pro-nobis (Pereskia aculeata Miller) leaves.
Cristina Oliveira Neves, Isabelle; Aparecida Rodrigues, Adrise; Teixeira Valentim, Thamires; Cristina Freitas de Oliveira Meira, Ana; Henrique Silva, Sérgio; Ayra Alcântara Veríssimo, Lizzy; Vilela de Resende, Jaime.
Afiliação
  • Cristina Oliveira Neves I; Department of Food Science, Federal University of Lavras, Lavras, Minas Gerais 37200-900, Brazil.
  • Aparecida Rodrigues A; Department of Food Science, Federal University of Lavras, Lavras, Minas Gerais 37200-900, Brazil.
  • Teixeira Valentim T; Department of Food Science, Federal University of Lavras, Lavras, Minas Gerais 37200-900, Brazil.
  • Cristina Freitas de Oliveira Meira A; Department of Food Science, Federal University of Lavras, Lavras, Minas Gerais 37200-900, Brazil.
  • Henrique Silva S; Department of Food Science, Federal University of Lavras, Lavras, Minas Gerais 37200-900, Brazil.
  • Ayra Alcântara Veríssimo L; Department of Food Science, Federal University of Lavras, Lavras, Minas Gerais 37200-900, Brazil. Electronic address: lizzy.alcantara@ufla.br.
  • Vilela de Resende J; Department of Food Science, Federal University of Lavras, Lavras, Minas Gerais 37200-900, Brazil.
J Chromatogr B Analyt Technol Biomed Life Sci ; 1161: 122435, 2020 Dec 15.
Article em En | MEDLINE | ID: mdl-33246278
The surfaces of the polyacrylamide cryogels were coated with L-tryptophan (cryogel-Trp) or L-phenylalanine (cryogel-Phe) to enhance crude leaf extract-derived ora-pro-nobis (OPN) protein binding via pseudo-specific hydrophobic interactions. Cryogels functionalized with amino acids were prepared and characterized through morphological, hydrodynamic, and thermal analyses. The adsorption capacities of cryogel-Phe and cryogel-Trp were evaluated in terms of type (sodium sulfate or sodium phosphate) and concentration (0.02 or 0.10 mol∙L-1) of saline solution, pH (4.0, 5.5, or 7.0), and NaCl concentration (0.0 or 0.5 mol∙L-1). The cryogel-Phe presented a higher adsorptive capacity, achieving its maximum value (q = 92.53 mg∙g-1) when the crude OPN crude leaf extract was diluted in sodium sulfate 0.02 mol∙L-1 + NaCl 0.50 mol∙L-1, at pH = 7.0. The dilution rate significantly (p < 0.05) affected the recovered protein amount after the adsorption and elution processes, reaching 94.45% when the feedstock solution was prepared with a crude extract 5 times. The zeta potential for the eluted OPN proteins was 5.76 mV (pH = 3.23) for both dilution rates. The secondary structure composition mainly included ß-sheets (46.50%) and α-helices (13.93%). The cryogel-Phe exhibited interconnected pores ranging 20-300 µm in size, with a Young modulus of 1.51 MPa, and thermal degradation started at 230 °C. These results indicate that the cryogel-Phe exhibited satisfactory properties as promising chromatography support for use in high-throughput purification of crude leaf extract-derived OPN proteins.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas de Plantas / Cromatografia de Afinidade / Cactaceae / Aminoácidos Aromáticos / Criogéis Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Assunto da revista: ENGENHARIA BIOMEDICA Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda
Texto completo
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas de Plantas / Cromatografia de Afinidade / Cactaceae / Aminoácidos Aromáticos / Criogéis Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Assunto da revista: ENGENHARIA BIOMEDICA Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda