Immobilisation of horseradish peroxidase on EupergitC for the enzymatic elimination of phenol.
In this study, three different approaches for the covalent immobilisation of the horseradish peroxidase (HRP) onto epoxy-activated acrylic polymers (EupergitC) were explored for the first time, direct HRP binding to the polymers via their oxirane groups, HRP binding to the polymers via a spacer made from adipic dihydrazide, and HRP binding to hydrazido polymer surfaces through the enzyme carbohydrate moiety previously modified by periodate oxidation. The periodate-mediated covalent immobilisation of the HRP on hydrazido EupergitC was found to be the most effective method for the preparation of biocatalysts. In this case, a maximum value of the immobilised enzyme activity of 127 U/g(support) was found using an enzyme loading on the support of 35.2mg/g(support). The free and the immobilised HRP were used to study the elimination of phenol in two batch reactors. As expected, the activity of the immobilised enzyme was lower than the activity of the free enzyme. Around 85% of enzyme activity is lost during the immobilisation. However, the reaction using immobilised enzyme showed that it was possible to reach high degrees of phenol removal (around 50%) using about one hundredth of the enzyme used in the soluble form.
Pramparo
L
L
Departament d'Enginyeria Quimica, Escola Tècnica Superior d'Enginyeria Química, Universitat Rovira i Virgili, Av Països Catalans 26, 43007 Tarragona, Spain.
Stüber
F
F
Font
J
J
Fortuny
A
A
Fabregat
A
A
Bengoa
C
C
D003432:Cross-Linking Reagents / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000378:metabolism*; D008722:Methods; D010504:Periodic Acid / Q000737:chemistry; D019800:Phenol / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry*; D011485:Protein Binding
D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D011108:Polymers; D010504:Periodic Acid; D019800:Phenol; C065843:Eupergit C; C009288:metaperiodate; D006735:Horseradish Peroxidase