Improvement of enzymatic performance of Asclepias curassavica L. proteases by immobilization. Application to the synthesis of an antihypertensive peptide
| Autor: | Gerardo Enrique Camí, Fanny Guzmán, Sonia Esther Barberis, Anabella Lucía Origone, Ronny Martinez, Andrés Illanes, Cintia Claudia Bernal, Constanza Silvina Liggieri |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Asclepias curassavica Proteases Immobilized enzyme Bioengineering Peptide INGENIERÍAS Y TECNOLOGÍAS 01 natural sciences Applied Microbiology and Biotechnology Biochemistry Biotecnología Industrial ANTIHYPERTENSIVE PEPTIDE 03 medical and health sciences 010608 biotechnology Reactivity (chemistry) 030304 developmental biology chemistry.chemical_classification 0303 health sciences PROTEASES IMMOBILIZATION biology ASCLEPIAS CURASSAVICA L ENZYMATIC SYNTHESIS OF PEPTIDES POROUS SILICA SUPPORTS Bioprocesamiento Tecnológico Biocatálisis Fermentación biology.organism_classification Enzyme chemistry Covalent bond Yield (chemistry) |
| Zdroj: | Process Biochemistry. 95:36-46 |
| ISSN: | 1359-5113 |
| DOI: | 10.1016/j.procbio.2020.05.013 |
| Popis: | The aim of this work was to study different immobilization strategies on silica supports in order to obtain robust biocatalysts from latex proteases of Asclepias curassavica L., a South American native plant. Immobilized enzyme performance was evaluated under harsh reaction conditions such as the synthesis of the antihypertensive peptide N-α-CBZ-Val-Gly-OH. Proteases from A. curassavica, named asclepain, were immobilized (0.51–5.56 mg of protein/ g of support) in non-functionalized silica (S), in glyoxyl-silica (GS) and in octyl-glyoxyl-silica (OGS), by adsorption, and multi-point covalent attachment on mono and hetero-functional supports, respectively, under previously determined optimal immobilization conditions. Immobilization yields were expressed as activity yield (Ya) and protein yield (Yp). Asclepain-OGS showed the highest Ya (178 ± 1.62 %) meaning an expressed activity 1.8 times higher than the offered activity, while Yp was 75 ± 0.4 %. Ya for asclepain-S and -GS were 64 ± 1.45 % and 16 ± 0.37 %, respectively. Best results were attributed to the ability of OGS support to guide the enzyme before covalent attachment, increasing its reactivity. Asclepain-OGS led to product yield of 95.5 ± 0.14 %, five times higher than soluble asclepain in the synthesis of N-α-CBZ-Val-Gly-OH, after 3 h in 30 % methanol in 0.1 M Tris-HCl buffer pH 8. Fil: Origone, Anabella Lucía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Física Aplicada "Dr. Jorge Andrés Zgrablich". Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto de Física Aplicada "Dr. Jorge Andrés Zgrablich"; Argentina. Universidad de La Serena; Chile. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Laboratorio de Bromatología; Argentina Fil: Barberis, Sonia Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Física Aplicada "Dr. Jorge Andrés Zgrablich". Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto de Física Aplicada "Dr. Jorge Andrés Zgrablich"; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Laboratorio de Bromatología; Argentina Fil: Illanes, Andrés. Pontificia Universidad Católica de Valparaíso. Escuela de Ingeniería Bioquímica; Chile Fil: Guzmán, Fanny. Pontificia Universidad Católica de Valparaíso; Chile Fil: Camí, Gerardo Enrique. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina Fil: Liggieri, Constanza Silvina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentina. Universidad Maimónides. Centro de Estudios Biomédicos, Básicos, Aplicados y Desarrollo. Departamento de Ciencias Biológicas y Biomédicas; Argentina Fil: Martínez, Ronny. Universidad de La Serena; Chile Fil: Bernal, Cintia Claudia. Universidad de La Serena; Chile |
| Databáze: | OpenAIRE |
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