Whole cell immobilized amperometric biosensor based on Saccharomyces cerevisiae for selective determination of vitamin B1 (thiamine).
A new amperometric whole cell biosensor based on Saccharomyces cerevisiae immobilized in gelatin was developed for selective determination of vitamin B1 (thiamine). The biosensor was constructed by using gelatin and crosslinking agent glutaraldehyde to immobilize S. cerevisiae cells on the Teflon membrane of dissolved oxygen (DO) probe used as the basic electrode system combined with a digital oxygen meter. The cells were induced by vitamin B1 in the culture medium, and the cells used it as a carbon source in the absence of glucose. So, when the vitamin B1 solution is injected into the whole cell biosensor system, an increase in respiration activity of the cells results from the metabolic activity and causes a decrease in the DO concentration of interval surface of DO probe related to vitamin B1 concentration. The response time of the biosensor is 3 min, and the optimal working conditions of the biosensor were carried out as pH 7.0, 50mM Tris-HCl, and 30 degrees C. A linear relationship was obtained between the DO concentration decrease and vitamin B1 concentration between 5.0 x 10(-3) and 10(-1) microM. In the application studies of the biosensor, sensitive determination of vitamin B1 in the vitamin tablets was investigated.
Akyilmaz
Erol
E
Department of Biochemistry, Faculty of Science, Ege University, 35100 Bornova-Izmir, Turkey. erol.akyilmaz@ege.edu.tr
Yaşa
Ihsan
I
Dinçkaya
Erhan
E
D015374:Biosensing Techniques / Q000379:methods*; D002138:Calibration; D004305:Dose-Response Relationship, Drug; D004563:Electrochemistry; D004867:Equipment Design; D005780:Gelatin / Q000737:chemistry; D005976:Glutaral / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D010100:Oxygen / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D013831:Thiamine / Q000032:analysis*; D013997:Time Factors
D005780:Gelatin; D010100:Oxygen; D005976:Glutaral; D013831:Thiamine