Conductometric Biosensor Based on Flavocytochrome b2 for L-Lactate Determination

2011 ◽  
Vol 9 (6) ◽  
pp. 2388-2391 ◽  
Author(s):  
O. Berezhetska ◽  
Y. Korpan ◽  
M. Gonchar
Keyword(s):  
Flavocytochrome B2 ◽  
Conductometric Biosensor

scholarly journals “Green” Prussian Blue Analogues as Peroxidase Mimetics for Amperometric Sensing and Biosensing

Biosensors ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 193
Author(s):  
Galina Z. Gayda ◽  
Olha M. Demkiv ◽  
Yanna Gurianov ◽  
Roman Ya. Serkiz ◽  
Halyna M. Klepach ◽  
...  
Keyword(s):  
Prussian Blue ◽  
Noble Metals ◽  
High Sensitivity ◽  
Graphite Electrodes ◽  
Flavocytochrome B2 ◽  
Prussian Blue Analogues ◽  
Scientific Investigations ◽  
Electrochemically Active ◽  
Amperometric Sensing ◽  
Prussian Blue Analogs

Prussian blue analogs (PBAs) are well-known artificial enzymes with peroxidase (PO)-like activity. PBAs have a high potential for applications in scientific investigations, industry, ecology and medicine. Being stable and both catalytically and electrochemically active, PBAs are promising in the construction of biosensors and biofuel cells. The “green” synthesis of PO-like PBAs using oxido-reductase flavocytochrome b2 is described in this study. When immobilized on graphite electrodes (GEs), the obtained green-synthesized PBAs or hexacyanoferrates (gHCFs) of transition and noble metals produced amperometric signals in response to H2O2. HCFs of copper, iron, palladium and other metals were synthesized and characterized by structure, size, catalytic properties and electro-mediator activities. The gCuHCF, as the most effective PO mimetic with a flower-like micro/nano superstructure, was used as an H2O2-sensitive platform for the development of a glucose oxidase (GO)-based biosensor. The GO/gCuHCF/GE biosensor exhibited high sensitivity (710 A M−1m−2), a broad linear range and good selectivity when tested on real samples of fruit juices. We propose that the gCuHCF and other gHCFs synthesized via enzymes may be used as artificial POs in amperometric oxidase-based (bio)sensors.

Functional replacement of yeast flavocytochrome b2 with bacterial l-lactate dehydrogenase

2010 ◽  
Vol 110 (3) ◽  
pp. 269-272 ◽  
Author(s):  
Hiroaki Sakai ◽  
Kazuhiro Nagahama ◽  
Hisataka Taguchi ◽  
Takashi Akamatsu ◽  
Shigeru Morimura ◽  
...  
Keyword(s):  
Lactate Dehydrogenase ◽  
Flavocytochrome B2 ◽  
Functional Replacement

Isolation and characterisation of the cytochrome domain of flavocytochrome b2 expressed independently in E. coli

1991 ◽  
Vol 43 (2-3) ◽  
pp. 312
Author(s):  
C.A. Brunt ◽  
M.C. Cox ◽  
A. Thurgood ◽  
G.R. Moore ◽  
G.A. Reid ◽  
...  
Keyword(s):  
E Coli ◽  
Flavocytochrome B2

scholarly journals The importance of the interdomain hinge in intramolecular electron transfer in flavocytochrome b2

1993 ◽  
Vol 291 (1) ◽  
pp. 89-94 ◽  
Author(s):  
P White ◽  
F D C Manson ◽  
C E Brunt ◽  
S K Chapman ◽  
G A Reid
Keyword(s):  
Electron Transfer ◽  
Steady State ◽  
Cytochrome C ◽  
Kinetic Properties ◽  
Stopped Flow ◽  
Wild Type ◽  
Wild Type Enzyme ◽  
Cytochrome C Reductase ◽  
Flavocytochrome B2 ◽  
Cytochrome C Reduction

The two distinct domains of flavocytochrome b2 (L-lactate:cytochrome c oxidoreductase) are connected by a typical hinge peptide. The amino acid sequence of this interdomain hinge is dramatically different in flavocytochromes b2 from Saccharomyces cerevisiae and Hansenula anomala. This difference in the hinge is believed to contribute to the difference in kinetic properties between the two enzymes. To probe the importance of the hinge, an interspecies hybrid enzyme has been constructed comprising the bulk of the S. cerevisiae enzyme but containing the H. anomala flavocytochrome b2 hinge. The kinetic properties of this ‘hinge-swap’ enzyme have been investigated by steady-state and stopped-flow methods. The hinge-swap enzyme remains a good lactate dehydrogenase as is evident from steady-state experiments with ferricyanide as acceptor (only 3-fold less active than wild-type enzyme) and stopped-flow experiments monitoring flavin reduction (2.5-fold slower than in wild-type enzyme). The major effect of the hinge-swap mutation is to lower dramatically the enzyme's effectiveness as a cytochrome c reductase; kcat. for cytochrome c reduction falls by more than 100-fold, from 207 +/- 10 s-1 (25 degrees C, pH 7.5) in the wild-type enzyme to 1.62 +/- 0.41 s-1 in the mutant enzyme. This fall in cytochrome c reductase activity results from poor interdomain electron transfer between the FMN and haem groups. This can be demonstrated by the fact that the kcat. for haem reduction in the hinge-swap enzyme (measured by the stopped-flow method) has a value of 1.61 +/- 0.42 s-1, identical with the value for cytochrome c reduction and some 300-fold lower than the value for the wild-type enzyme. From these and other kinetic parameters, including kinetic isotope effects with [2-2H]lactate, we conclude that the hinge plays a crucial role in allowing efficient electron transfer between the two domains of flavocytochrome b2.

Sol–gel-derived thick-film conductometric biosensor for urea determination in serum

2000 ◽  
Vol 404 (2) ◽  
pp. 195-203 ◽  
Author(s):  
Won-Yong Lee ◽  
Seung-Ryeol Kim ◽  
Tae-Han Kim ◽  
Kang Shin Lee ◽  
Min-Chol Shin ◽  
...  
Keyword(s):  
Thick Film ◽  
Sol Gel ◽  
Urea Determination ◽  
Conductometric Biosensor

Kinetic testing of a hypothetical complex between flavocytochrome b2 and cytochrome c

1995 ◽  
Vol 59 (2-3) ◽  
pp. 438
Author(s):  
D.M. Short ◽  
F.D.C. Manson ◽  
G.A. Reid ◽  
S.K. Chapman
Keyword(s):  
Cytochrome C ◽  
Flavocytochrome B2 ◽  
Kinetic Testing

Altering the Electron Acceptor Specificity of Flavocytochrome b2

1999 ◽  
Vol 27 (1) ◽  
pp. A45-A45
Author(s):  
Fraser Welsh ◽  
Stuart Rivers ◽  
Stephen K. Chapman ◽  
Graeme A. Reid
Keyword(s):  
Electron Acceptor ◽  
Flavocytochrome B2 ◽  
Acceptor Specificity
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