Akyilmaz Erol, Yaşa Ihsan, Dinçkaya Erhan
Department of Biochemistry, Faculty of Science, Ege University, 35100 Bornova-Izmir, Turkey.
Anal Biochem. 2006 Jul 1;354(1):78-84. doi: 10.1016/j.ab.2006.04.019. Epub 2006 May 2.
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.
一种基于固定在明胶中的酿酒酵母的新型安培型全细胞生物传感器被开发用于选择性测定维生素B1(硫胺素)。该生物传感器通过使用明胶和交联剂戊二醛将酿酒酵母细胞固定在用作基本电极系统的溶解氧(DO)探头的聚四氟乙烯膜上,并与数字式氧气计结合构建而成。细胞在培养基中由维生素B1诱导,并且在没有葡萄糖的情况下细胞将其用作碳源。因此,当将维生素B1溶液注入全细胞生物传感器系统时,细胞呼吸活性的增加源于代谢活性,并导致与维生素B1浓度相关的DO探头间隔表面的DO浓度降低。该生物传感器的响应时间为3分钟,其最佳工作条件为pH 7.0、50mM Tris-HCl和30℃。在5.0×10(-3)至10(-1) microM之间,DO浓度降低与维生素B1浓度之间获得了线性关系。在该生物传感器的应用研究中,对维生素片中维生素B1的灵敏测定进行了研究。