| Autor: |
Schuhmann Wolfgang, Dmytruk Olena V, Smutok Oleh V, Dmytruk Kostyantyn V, Sibirny Andriy A |
| Jazyk: |
angličtina |
| Rok vydání: |
2011 |
| Předmět: |
|
| Zdroj: |
BMC Biotechnology, Vol 11, Iss 1, p 58 (2011) |
| Druh dokumentu: |
article |
| ISSN: |
1472-6750 |
| DOI: |
10.1186/1472-6750-11-58 |
| Popis: |
Abstract Background The detection and quantification of uric acid in human physiological fluids is of great importance in the diagnosis and therapy of patients suffering from a range of disorders associated with altered purine metabolism, most notably gout and hyperuricaemia. The fabrication of cheap and reliable urate-selective amperometric biosensors is a challenging task. Results A urate-selective microbial biosensor was developed using cells of the recombinant thermotolerant methylotrophic yeast Hansenula polymorpha as biorecognition element. The construction of uricase (UOX) producing yeast by over-expression of the uricase gene of H. polymorpha is described. Following a preliminary screening of the transformants with increased UOX activity in permeabilized yeast cells the optimal cultivation conditions for maximal UOX yield namely a 40-fold increase in UOX activity were determined. The UOX producing cells were coupled to horseradish peroxidase and immobilized on graphite electrodes by physical entrapment behind a dialysis membrane. A high urate selectivity with a detection limit of about 8 μM was found. Conclusion A strain of H. polymorpha overproducing UOX was constructed. A cheap urate selective microbial biosensor was developed. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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