Epoxy Sepabeads: A Novel Epoxy Support for Stabilization of Industrial Enzymes via Very Intense Multipoint Covalent Attachment

2002 ◽  
Vol 18 (3) ◽  
pp. 629-634 ◽  
Author(s):  
C. Mateo ◽  
O. Abian ◽  
G. Fernandez-Lorente ◽  
J. Pedroche ◽  
R. Fernandez-Lafuente ◽  
...  
Keyword(s):  
Covalent Attachment ◽  
Industrial Enzymes ◽  
Multipoint Covalent Attachment

Advances in the design of new epoxy supports for enzyme immobilization–stabilization

2007 ◽  
Vol 35 (6) ◽  
pp. 1593-1601 ◽  
Author(s):  
C. Mateo ◽  
V. Grazú ◽  
B.C.C. Pessela ◽  
T. Montes ◽  
J.M. Palomo ◽  
...  
Keyword(s):  
Physical Adsorption ◽  
Immobilized Enzymes ◽  
Covalent Immobilization ◽  
Covalent Attachment ◽  
Industrial Enzymes ◽  
Protein Surface ◽  
Strong Stabilization ◽  
Support Surfaces ◽  
Multipoint Covalent Attachment ◽  
Epoxy Groups

Multipoint covalent immobilization of enzymes (through very short spacer arms) on support surfaces promotes a very interesting ‘rigidification’ of protein molecules. In this case, the relative positions of each residue of the enzyme involved in the immobilization process have to be preserved unchanged during any conformational change induced on the immobilized enzyme by any distorting agent (heat, organic solvents etc.). In this way, multipoint covalent immobilization should induce a very strong stabilization of immobilized enzymes. Epoxy-activated supports are able to chemically react with all nucleophile groups placed on the protein surface: lysine, histidine, cysteine, tyrosine etc. Besides, epoxy groups are very stable. This allows the performance of very long enzyme–support reactions, enabling us to get very intense multipoint covalent attachment. In this way, these epoxy supports seem to be very suitable to stabilize industrial enzymes by multipoint covalent attachment. However, epoxy groups exhibit a low intermolecular reactivity towards nucleophiles and hence the enzymes are not able to directly react with the epoxy supports. Thus a rapid physical adsorption of enzymes on the supports becomes a first step, followed by an additional rapid ‘intramolecular’ reaction between the already adsorbed enzyme and the activated support. In this situation, a suitable first orientation of the enzyme on the support (e.g. through regions that are very rich in nucleophiles) is obviously necessary to get a very intense additional multipoint covalent immobilization. The preparation of different ‘generations’ of epoxy supports and the design of different protocols to fully control the first interaction between enzymes and epoxy supports will be reviewed in this paper. Finally, the possibilities of a directed immobilization of mutated enzymes (change of an amino acid by cysteine on specific points of the protein surface) on tailor-made disulfide-epoxy supports will be discussed as an almost-ideal procedure to achieve very intense and very efficient rigidification of a desired region of industrial enzymes.

The presence of thiolated compounds allows the immobilization of enzymes on glyoxyl agarose at mild pH values: New strategies of stabilization by multipoint covalent attachment

2009 ◽  
Vol 45 (6-7) ◽  
pp. 477-483 ◽  
Author(s):  
Juan M. Bolivar ◽  
Fernando López-Gallego ◽  
Cesar Godoy ◽  
Dasciana S. Rodrigues ◽  
Rafael C. Rodrigues ◽  
...  
Keyword(s):  
Covalent Attachment ◽  
Ph Values ◽  
Immobilization Of Enzymes ◽  
Multipoint Covalent Attachment ◽  
New Strategies

Immobilization and Stabilization of Proteins by Multipoint Covalent Attachment on Novel Amino-Epoxy-Sepabeads®

2006 ◽  
pp. 153-162 ◽  
Author(s):  
Cesar Mateo ◽  
Benevides C. C. Pessela ◽  
Valeria Grazu ◽  
Fernando López-Gallego ◽  
Rodrigo Torres ◽  
...  
Keyword(s):  
Covalent Attachment ◽  
Multipoint Covalent Attachment

Stabilization by multipoint covalent attachment of a biocatalyst with polygalacturonase activity used for juice clarification

2016 ◽  
Vol 208 ◽  
pp. 252-257 ◽  
Author(s):  
Yuly A. Ramírez Tapias ◽  
Cintia W. Rivero ◽  
Fernando López Gallego ◽  
José M. Guisán ◽  
Jorge A. Trelles
Keyword(s):  
Covalent Attachment ◽  
Juice Clarification ◽  
Multipoint Covalent Attachment ◽  
Polygalacturonase Activity

Hyperstabilization of a thermophilic esterase by multipoint covalent attachment

1995 ◽  
Vol 17 (4) ◽  
pp. 366-372 ◽  
Author(s):  
R. Fernandez-Lafuente ◽  
D.A. Cowan ◽  
A.N.P. Wood
Keyword(s):  
Covalent Attachment ◽  
Thermophilic Esterase ◽  
Multipoint Covalent Attachment

scholarly journals Co-Immobilization and Co-Localization of Oxidases and Catalases: Catalase from Bordetella Pertussis Fused with the Zbasic Domain

Catalysts ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 810
Author(s):  
Paz García-García ◽  
Javier Rocha-Martin ◽  
Jose M. Guisan ◽  
Gloria Fernandez-Lorente
Keyword(s):  
Hydrogen Peroxide ◽  
Bordetella Pertussis ◽  
Specific Activity ◽  
Amine Oxidase ◽  
Bovine Liver ◽  
Exchange Chromatography ◽  
Covalent Attachment ◽  
Opposite Hemisphere ◽  
Cationic Exchange ◽  
Multipoint Covalent Attachment

Oxidases catalyze selective oxidations by using molecular oxygen as an oxidizing agent. This process promotes the release of hydrogen peroxide, an undesirable byproduct. The instantaneous elimination of hydrogen peroxide can be achieved by co-immobilization and co-localization of the oxidase and an auxiliary catalase inside the porous structure of solid support. In this paper, we proposed that catalase from Bordetella pertussis fused with a small domain (Zbasic) as an excellent auxiliary enzyme. The enzyme had a specific activity of 23 U/mg, and this was almost six-fold higher than the one of the commercially available catalases from bovine liver. The Zbasic domain was fused to the four amino termini of this tetrameric enzyme. Two domains were close in one hemisphere of the enzyme molecule, and the other two were close in the opposite hemisphere. In this way, each hemisphere contained 24 residues with a positive charge that was very useful for the purification of the enzyme via cationic exchange chromatography. In addition to this, each hemisphere contained 10 Lys residues that were very useful for a rapid and intense multipoint covalent attachment on highly activated glyoxyl supports. In fact, 190 mg of the enzyme was immobilized on one gram of glyoxyl-10% agarose gel. The ratio catalase/oxidase able to instantaneously remove more than 93% of the released hydrogen peroxide was around 5–6 mg of catalase per mg of oxidase. Thirty milligrams of amine oxidase and 160 mg of catalase were co-immobilized and co-localized per gram of glyoxyl-agarose 10BCL (10% beads cross-linked) support. This biocatalyst eliminated biogenic amines (putrescine) 80-fold faster than a biocatalyst of the same oxidase co-localized with the commercial catalase from bovine liver.

Immobilization of Proteins on Glyoxyl Activated Supports: Dramatic Stabilization of Enzymes by Multipoint Covalent Attachment on Pre-Existing Supports

2015 ◽  
Vol 19 (999) ◽  
pp. 1-1 ◽  
Author(s):  
Gloria Fernandez-Lorente ◽  
Fernando Lopez-Gallego ◽  
Juan M. Bolivar ◽  
Javier Rocha-Martin ◽  
Sonia Moreno-Perez ◽  
...  
Keyword(s):  
Covalent Attachment ◽  
Multipoint Covalent Attachment
Sign in / Sign up

Export Citation Format

Share Document