Stabilization of enzymes by multipoint covalent immobilization on supports activated with glyoxyl groups.
Stabilization of enzymes via immobilization techniques is a valuable approach in order to convert a necessary protocol (immobilization) into a very interesting tool to improve key enzyme properties (stabilization). Multipoint covalent attachment of each immobilized enzyme molecule may promote a very interesting stabilizing effect. The relative distances among all enzyme residues involved in immobilization has to remain unaltered during any conformational change induced by any distorting agent. Amino groups are very interesting nucleophiles placed on protein surfaces. The immobilization of enzyme through the region having the highest amount of amino groups (Lys residues) is key for a successful stabilization. Glyoxyl groups are small aliphatic aldehydes that form very unstable Schiff's bases with amino groups and they do not seem to be useful for enzyme immobilization at neutral pH. However, under alkaline conditions, glyoxyl supports are able to immobilize enzymes via a first multipoint covalent immobilization through the region having the highest amount of Lysine groups. Activation of supports with a high surface density of glyoxyl groups and the performance of very intense enzyme-support multipoint covalent attachments are here described.
Institute of Catalysis, CSIC, CAMPUS UAM-Cantoblanco, Madrid, Spain.
Fernando
F
López-Gallego
Gloria
G
Fernandez-Lorente
Javier
J
Rocha-Martin
Juan M
JM
Bolivar
Cesar
C
Mateo
Jose M
JM
Guisan
D000588:Amines / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D006038:Glyoxylates / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000737:chemistry; D010399:Penicillin Amidase / Q000737:chemistry*; D011485:Protein Binding; D012545:Schiff Bases / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry*
D000588:Amines; D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D029968:Escherichia coli Proteins; D006038:Glyoxylates; D012545:Schiff Bases; C070399:glyoxyl agarose; D012685:Sepharose; D010399:Penicillin Amidase; D008239:Lysine