Amperometric Biosensors for Detection of Sugars Based on the Electrical Wiring of Different Pyranose Oxidases and Pyranose Dehydrogenases with Osmium Redox Polymer on Graphite Electrodes

Abstract

Electrical wiring of different types of pyranose oxidase (P2O) (fungal wild type, recombinant wild type with a hexahistidine tag, mutant form E542K with a hexa-histidine tag) from Trametes multicolor, and recombinant P2O from Coriolus sp. overexpressed in Escherichia coli as well as of pyranose dehydrogenase (PDH) from Agaricus meleagris and Agaricus xanthoderma with an osmium redox polymer (poly(1-vinylimidazole) 12 -[Os(4,4'-dimethyl-2,2'-dipyridyl) 2 -Cl 2 ] 2þ/þ ) on graphite electrodes was carried out. After optimization studies using glucose as substrate, the biosensors, which showed the best characteristics in terms of linear range, detection limit and sensitivity were selected, viz. wild type P2O from T. multicolor and PDH from A. meleagris. These two enzymes were used and investigated for their selectivity for a number of different sugars.

Engineering of pyranose 2-oxidase for modified oxygen reactivity

“…Pyranose 2-oxidase is a well investigated oxidoreductase for industrial applications in the area of food technology and biosensors/biofuel cells [14,[52][53][54][55]. Reduced oxygen reactivity is a desirable property for some applications because of reduced The reactivity of flavoproteins with oxygen is of considerable scientific interest, and the determinants of oxygen activation and reactivity are the subject of numerous studies [39][40][41][42][43][44].…”

Offline glucose biomonitoring in yeast culture by polyamidoamine/ cysteamine‐modified gold electrodes

“…Different to GOx, this enzyme catalysis the glucose at carbon-2 with high affinity and generates 2-keto sugars and H 2 O 2 as shown below 28 :…”

Engineered Pyranose 2‐Oxidase: Efficiently Turning Sugars into Electrical Energy

“…As judged from the catalytic efficiency, some of these alternative electron acceptors are in fact better substrates for the enzyme than oxygen, suggesting that P2Ox can play an important role in the reduction of quinones during the process of ligninolysis in vivo [20,21]. This excellent reactivity of P2Ox with alternative electron acceptors and a range of different sugar substrates, like dglucose, d-galactose and d-xylose, can be employed for applications in enzymatic biosensors and BFCs, which has already been shown before [5]. There are two main advantages when using P2Ox in BFCs rather than GOx, namely the lack of any anomeric preference and the much broader substrate spectrum, which is useful for small scale energy production from e.g.…”

“…Samples were injected with an injector (Rheodyne, type 7125 LabPRO, Cotati, CA, USA) supplied with an injection loop of 100 mL. A 50 mM ethanolamine buffer at pH 8.5 was used as the carrier buffer in the flow system for test-runs with the Os redox polymer and a 50 mM KH 2 PO 4 -buffer at pH 6.5 for analyses with 1,4-BQ [5]. The carrier buffer was thoroughly degassed under vacuum to prevent microbubbles to appear in the flow system and pumped at a flow rate of 0.5 mL · min À1 (Minipuls 2, Gilson, Villier-le Bel, France).…”

“…The transport of the electrons can occur directly by coupling the enzyme to the electrodes, in a process called direct electron transfer (DET) [2], however only a very restricted number of oxidoreductases show facile DET properties [3]. In mediated electron transfer, on the other hand, certain redox active compounds, denoted as mediators, such as quinones [4], ferrocenes or osmium redox polymers [5], largely facilitate electron transfer between the active site of the enzyme and the electrode through collecting the electrons from the prosthetic group of the enzyme and transferring them to an electrode of suitable material.…”