Construction of uricase-overproducing strains of Hansenula polymorpha and its application as biological recognition element in microbial urate biosensor.
BMC Biotechnol
; 11: 58, 2011 May 25.
Article
em En
| MEDLINE
| ID: mdl-21612631
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Pichia
/
Urato Oxidase
/
Ácido Úrico
/
Técnicas Biossensoriais
Limite:
Humans
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article