Effect of amine length in the interference of the multipoint covalent immobilization of enzymes on glyoxyl agarose beads
| Autor: | Justo Pedroche, Roberto Fernandez-Lafuente, El-Hocine Siar, Mª del Carmen Millán, Roberto Morellon-Sterling, Diandra de Andrades, Sabrina Ait Braham |
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| Přispěvatelé: | Ministerio de Ciencia e Innovación (España), Universidad de Sevilla. Departamento de Bioquímica Médica y Biología Molecular e Inmunología, Ministerio de Ciencia e Innovacion CTQ2017-86170-R, Ministerio de Educación |
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
0301 basic medicine Steric effects Immobilized enzyme Multipoint covalent attachment Bioengineering 01 natural sciences Applied Microbiology and Biotechnology Aldehyde 03 medical and health sciences 010608 biotechnology Enzyme Stability medicine Organic chemistry Enzyme immobilization Steric hindrances Glyoxyl agarose Amines chemistry.chemical_classification Chymotrypsin biology Sepharose Glyoxylates Enzyme stabilization General Medicine Hydrogen-Ion Concentration Trypsin Enzymes Immobilized 030104 developmental biology Enzyme chemistry Covalent bond biology.protein Amine gas treating Biotechnology medicine.drug |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | 1 Tabla.-- 17 Figuras Trypsin, chymotrypsin, penicillin G acylase and ficin extract have been stabilized by immobilization on glyoxyl agarose, adding different aliphatic compounds bearing a primary amine group during the immobilization: ethyl amine, butyl amine, hexyl amine (at concentrations ranging from 0 to 20 mM) and octyl amine (from 0 to 10 mM) to analyze their effects on the immobilized enzyme stability. As expected, the presence of amines reduced the intensity of the enzyme-support multipoint covalent attachment, and therefore the enzyme stability. However, it is clear that this effect is higher using octyl amine for all enzymes (in some cases the enzyme immobilized in the presence of 10 mM octyl amine was almost inactivated while the reference kept over 50 % of the initial activity). This way, it seems that the most important effect of the presence of aminated compounds came from the generation of steric hindrances to the enzyme/support multi-reaction promoted by the ammines that are interacting with the aldehyde groups. In some instances, just 1 mM of aminated compounds is enough to greatly decrease enzyme stability. The results suggested that, if the composition of the enzyme extract is unknown, to eliminate small aminated compounds may be necessary to maximize the enzyme-support reaction. We gratefully recognize the financial support from Ministerio de Ciencia e Innovación-Spanish Government (project number CTQ2017-86170-R). RMS thank to Ministerio de Educacion -Spanish Government for a FPU fellowship, SAB and EHS thank Algerian Ministry of higher education and scientific research for their fellowships. The help and suggestions from Dr. Ángel Berenguer (Departamento de Química Inorgánica, Universidad de Alicante) are gratefully recognized. |
| Databáze: | OpenAIRE |
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