RESUMO
A microbial biosensor based on thick film technology was developed. The microorganisms, Arthrobacter nicotianae, were immobilized in Ca-alginate directly on the electrode surface. For the stability of the calcium alginate gel the addition of 0.5 mM CaCl2 to the assay buffer was necessary. The respiratory activity of the microorganisms was monitored by oxygen consumption at -600 mV vs. Ag/AgCl reference electrode. The sensor was used in a batch system and was applied to the determination of free fatty acids in milk. Short-chain fatty acids (C4:0-C12:0) were the preferential substrates, with butyric acid being the main substrate. Consequently, the concentration of free short-chain fatty acids was represented as the butyric acid equivalent. The sensor showed linearity over the concentration range 9.5-165.5 microM (correlation coefficient, r = 0.99920). The response time of the sensor was approximately 3 min. No additional dialysis membrane was necessary, which led to a high sensitivity of the sensor and fast response times. Recovery rates of 98-113% were found for butyric acid in milk samples using the sensor without any additional membrane and a sample dilution of 200 by the assay. Two widespread disadvantages of microbial sensors, long response times and long times to return to the baseline signal after use, could be overcome.
