The orientationally controlled assembly of genetically modified enzymes in an amperometric biosensor

A kinetic analysis of three modified novel nitroreductases

“…Constructed plasmids were transformed into E. coli Rosetta pLysS (DE3k) host cells for protein overexpression and purified as described previously (Gwenin et al 2007). Purifications were visualised by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) 12 % w/v gels in a Lamelli buffer system using a BIO-RAD Protean 3 electrophoresis apparatus.…”

“…Increasing awareness of the threat of terrorist activity to national and global security has led to an urgent need to provide a rapid screening system for the detection of explosive compounds. In response an amperometric ''nanodog'' biosensor has been developed that exploits the properties of an immobilised E. coli K12 NfsB NTR, to facilitate explosive vapour detection in the part per trillion range (ppt) (Gwenin et al 2007(Gwenin et al , 2008. The ability of the sensor to differentiate between different explosive compounds by using an array of enzymes each with a discernible signature response is under development.…”

“…)cys 6 that has 6 cysteine codons inserted into the vector structure upstream of its multi-cloning site and downstream of the histidine tags. This vector has been constructed to facilitate enzyme immobilisation to form the biosensor (Gwenin et al 2007). The three enzymes were over-expressed from E. coli Rosetta (pLysS DE3k) with and without cysteine tags, purified, and characterised.…”

Enzyme‐Based Biosensors: Synthesis and Applications

“…Apart from the toxic compounds mentioned above, enzyme -based biosensors are also extensively used to monitor other some important pollutants, including nitroaromatic compounds [173,174] , heavy metals [31] , and typical infectious agents [42,134] , and have achieved considerable success. However, poor stability of the biosensor and short lifetime, mainly caused by electrode fouling and enzyme inactivation and desorption from immobilization supports, are both very important limitations because such challenges will not only impede biosensor transport, but also increase per -measurement cost.…”

Synthesis High Sensivity ZnO Cholesterol Biosensor with One Dimensional Nanostructures

“…Neurohumours of the neurotransmitter systems are also involved in conveying tactile signals [1]. The development of biosensors started in 1962, and it was based on the enzyme electrode concept proposed by Clark and Lyons [2]. This enzyme electrode concept is about fixing glucose oxidase (GOD) on a dissolved oxygen electrode, once the GOD contacts with glucose, the glucose will be oxidized.…”