title: Methylamine-sensitive amperometric biosensor based on (His)6-tagged Hansenula polymorpha methylamine oxidase immobilized on the gold nanoparticles.
abstract: A novel methylamine-selective amperometric bienzyme biosensor based on recombinant primary amine oxidase isolated from the recombinant yeast strain Saccharomyces cerevisiae and commercial horseradish peroxidase is described. Two amine oxidase preparations were used: free enzyme (AMO) and covalently immobilized on the surface of gold nanoparticles (AMO-nAu). Some bioanalytical parameters (sensitivity, selectivity, and storage stability) of the developed biosensors were investigated. The sensitivity for both sensors is high: 1450 ± 113 and 700 ± 30 A(-1) ·M(-1) ·m(-2) for AMO-nAu biosensor, respectively. The biosensors exhibit the linear range from 15 μM to 150 μM (AMO-nAu) and from 15 μM to 60 μM (AMO). The developed biosensor demonstrated a good selectivity toward methylamine (MA) (signal for dimethylamine and trimethylamine is less than 5% and for ethylamine 15% compared to MA output) and reveals a satisfactory storage stability. The constructed amperometric biosensor was used for MA assay in real samples of fish products in comparison with chemical method. The values obtained with both approaches different methods demonstrated a high correlation.
authors.affiliation: Institute of Cell Biology, NAS of Ukraine, Drahomanov Street 14/16, Lviv 79005, Ukraine.
authors.forename: Nataliya Ye
authors.initials: NY
authors.lastname: Stasyuk
authors.affiliation: Institute of Cell Biology, NAS of Ukraine, Drahomanov Street 14/16, Lviv 79005, Ukraine.
authors.forename: Oleh V
authors.initials: OV
authors.lastname: Smutok
authors.affiliation: Institute of Cell Biology, NAS of Ukraine, Drahomanov Street 14/16, Lviv 79005, Ukraine.
authors.forename: Andriy E
authors.initials: AE
authors.lastname: Zakalskiy
authors.affiliation: Institute of Cell Biology, NAS of Ukraine, Drahomanov Street 14/16, Lviv 79005, Ukraine.
authors.forename: Oksana M
authors.initials: OM
authors.lastname: Zakalska
authors.affiliation: Institute of Cell Biology, NAS of Ukraine, Drahomanov Street 14/16, Lviv 79005, Ukraine ; Institute of Applied Biotechnology and Basic Sciences, University of Rzeszow, Sokolowska Street 26, 36-100 Kolbuszowa, Poland.
authors.forename: Mykhailo V
authors.initials: MV
authors.lastname: Gonchar
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D004800:Enzymes, Immobilized / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry*; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D008744:Methylamines / Q000032:analysis*; D010088:Oxidoreductases / Q000737:chemistry*; D010843:Pichia / Q000201:enzymology*
chemical_list: D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D008744:Methylamines; D006046:Gold; C027451:methylamine; D010088:Oxidoreductases