title: Selective oxidation of glycerol to 1,3-dihydroxyacetone by covalently immobilized glycerol dehydrogenases with higher stability and lower product inhibition.
abstract: Glycerol dehydrogenase (GlyDH) catalyzes the regioselective oxidation of glycerol to yield 1,3-dihydroxyacetone (DHA); an important building block in chemical industry. Three recombinant GlyDHs from Geobacillus stearothermophilus, from Citrobacter braakii and from Cellulomonas sp. were stabilized by covalent immobilization. The highest activity recoveries (40-50%) of the insoluble preparations were obtained by immobilizing these enzymes in presence of polyethylene glycol (PEG). Noteworthy, these immobilized preparations were more stable and less inhibited by DHA than their soluble counterparts. In particular, GlyDH from G.stearothermophilus immobilized on agarose activated with both amine and glyoxyl groups and crosslinked with dextran aldehyde was 3.7-fold less inhibited by DHA than its soluble form and retained 100% of its initial activity after 18h of incubation at 65°C and pH 7. This is one of the few examples where the same immobilization protocol has minimized enzyme product inhibition and maximized thermal stability.
authors.affiliation: Departamento de Biocatálisis, Instituto de Catálisis y Petroleoquímica-CSIC, Campus UAM, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Rocha-Martin
authors.affiliation: Departamento de Biocatálisis, Instituto de Catálisis y Petroleoquímica-CSIC, Campus UAM, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Andreína
authors.initials: A
authors.lastname: Acosta
authors.affiliation: Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Departamento de Biología Molecular, Campus UAM, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Jose
authors.initials: J
authors.lastname: Berenguer
authors.affiliation: Departamento de Biocatálisis, Instituto de Catálisis y Petroleoquímica-CSIC, Campus UAM, Cantoblanco, 28049 Madrid, Spain. Electronic address: jmguisan@icp.csic.es.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Guisan
authors.affiliation: Biofunctional Nanomaterials Unit, CIC BiomaGUNE, Paseo Miramon 182, 20009 San Sebasitan, Spain; IKERBASQUE, Basque Foundation for Science, Bilbao, Spain. Electronic address: flopez.ikerbasque@cicbiomagune.es.
authors.forename: Fernando
authors.initials: F
authors.lastname: Lopez-Gallego
mesh_terms: D040143:Cellulomonas / Q000201:enzymology; D002954:Citrobacter / Q000201:enzymology; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D017931:DNA Primers / Q000235:genetics; D004098:Dihydroxyacetone / Q000096:biosynthesis* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004800:Enzymes, Immobilized / Q000378:metabolism; D004926:Escherichia coli; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D005990:Glycerol / Q000378:metabolism*; D007218:Industrial Microbiology / Q000379:methods*; D020128:Inhibitory Concentration 50; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D011092:Polyethylene Glycols
chemical_list: D003432:Cross-Linking Reagents; D017931:DNA Primers; D004800:Enzymes, Immobilized; D011092:Polyethylene Glycols; D010088:Oxidoreductases; D004098:Dihydroxyacetone; D005990:Glycerol
keywords: Dihydroxyacetone and protein stabilization; Enzyme immobilization; Glycerol dehydrogenase