title: His6-OPH enzyme-based bio-hybrid material for organophosphate detection.
abstract: In this work, we report on the development of a bio-sensing film for the detection of organophosphorous compounds using sol-gel technology. A novel sol-gel immobilization method employing tetraethoxysilane/3-glycidoxypropyltrimethoxysilane/water hybrid material was developed and used to immobilize the hexahistidine-tagged organophosphorous hydrolase enzyme (His(6)-OPH). Bio-sensing layers with encapsulated His(6)-OPH of various structures (water/silane, precursor ratios) have been prepared. The optimal (P = 5:1, R = 188) bio-sensing layers retained 90% of the initial enzyme activity. Furthermore, the bio-sensing layer prepared by this method was able to maintain its activity at or above 80% of its initial activity for 2 weeks. The bio-hybrid film also showed excellent reusability and improved activity at neutral pH in comparison to the same enzyme in solution.
authors.affiliation: Faculty of Mechanical Engineering, Centre of Sensor Technology, University of Maribor, Smetanova 17, 2000 Maribor, Slovenia.
authors.forename: Nina
authors.initials: N
authors.lastname: Frančič
authors.forename: Aljoša
authors.initials: A
authors.lastname: Košak
authors.forename: Ilya
authors.initials: I
authors.lastname: Lyagin
authors.forename: Elena N
authors.initials: EN
authors.lastname: Efremenko
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Lobnik
mesh_terms: D043303:Aryldialkylphosphatase / Q000737:chemistry / Q000378:metabolism*; D015374:Biosensing Techniques / Q000379:methods*; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D006639:Histidine / Q000737:chemistry / Q000378:metabolism*; D007306:Insecticides / Q000032:analysis; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D009943:Organophosphorus Compounds / Q000032:analysis*; D010261:Paraoxon / Q000032:analysis; D044367:Phase Transition; D012680:Sensitivity and Specificity
chemical_list: D004800:Enzymes, Immobilized; C471213:His-His-His-His-His-His; D007306:Insecticides; D009842:Oligopeptides; D009943:Organophosphorus Compounds; D006639:Histidine; D043303:Aryldialkylphosphatase; D010261:Paraoxon