Effect of the support and experimental conditions in the intensity of the multipoint covalent attachment of proteins on glyoxyl-agarose supports: Correlation between enzyme–support linkages and thermal stability

2007 ◽  
Vol 40 (5) ◽  
pp. 1160-1166 ◽  
Author(s):  
Justo Pedroche ◽  
Maria del Mar Yust ◽  
Cesar Mateo ◽  
Roberto Fernández-Lafuente ◽  
Julio Girón-Calle ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Covalent Attachment ◽  
Experimental Conditions ◽  
Multipoint Covalent Attachment ◽  
Effect Of The Support

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

scholarly journals A two-jet arc plasma: matrix effects and ways to their suppression

2019 ◽  
Vol 85 (1II)) ◽  
pp. 139-144
Author(s):  
N. P. Zaksas ◽  
A. F. Veryaskin
Keyword(s):  
Thermal Stability ◽  
Particle Size ◽  
Biological Samples ◽  
Matrix Effects ◽  
Certified Reference Materials ◽  
Organic Matrix ◽  
Graphite Powder ◽  
Powdered Sample ◽  
Experimental Conditions ◽  
Emission Analysis

A two-jet plasma is used for direct atomic emission analysis of powdered samples. It is characterized by relatively weak matrix effects, which allows using unified calibration samples on the basis of graphite powder for analysis of the samples with inorganic, organic, and organomineral matrix. In the present paper the effects limiting the usage of the unified approach due to different thermal stability and evaporation efficiency of the samples are discussed. The understated concentrations of a set of elements (Al, Ba, Ca, La, Mg, Mn, Sr, Ti, and Y) were obtained in analysis of certified reference materials of geological samples. It was shown that determination of rare earth elements should be carried out in the region behind the jet confluence providing their complete evaporation. For other elements, registration of the spectra in this region improves the results to some extent but they do not achieve the certified values. To speed up evaporation of these elements, the experimental conditions were chosen for plasma chemical reactions which provide conversion of the matrix elements into more volatile compounds. Addition of ammonium hydrofluoride to powdered sample considerably increased the line intensities of Al and Ca strongly associated with the silicon matrix. Incomplete evaporation was observed in analysis of biological samples with particle size more than 100 μm. A decrease in consumption of carrier argon is quite enough for effective decomposition of the organic matrix in plasma; the value of gas consumption depends on thermal stability and particle size of the sample. Preliminary sample carbonization is another way to improve evaporation of biological samples.

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

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.

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