Microscale biosensor for measurement of volatile fatty acids in anoxic environments

A microscale biosensor for acetate, propionate, isobutyrate, and lactate is described. The sensor is based on the bacterial respiration of low-molecular-weight, negatively charged species with a concomitant reduction of NO3- to N2O. A culture of denitrifying bacteria deficient in N2O reductase was immobilized in front of the tip of an electrochemical N2O microsensor. The bacteria were separated from the outside environment by an ion-permeable membrane and supplied with nutrients (except for electron donors) from a medium reservoir behind the N2O sensor. The signal of the sensor, which corresponded to the rate of N2O production, was proportional to the supply of the electron donor to the bacterial mass. The selectivity for volatile fatty acids compared to other organic compounds was increased by selectively enhancing the transport of negatively charged compounds into the sensor by electrophoretic migration (electrophoretic sensitivity control). The sensor was susceptible to interference from O-2, N2O, NO2-, H2S, and NO3-. Interference from NO3- was low and could be quantified and accounted for. The detection limit was equivalent to about I muM acetate, and the 90% response time was 30 to 90 s. The response of the sensor was not affected by changes in pH between 5.5 and 9 and was also unaffected by changes in salinity in the range of 2 to 32parts per thousand. The functioning of the sensor over a temperature span of 7 to 30degreesC was investigated. The concentration range for a linear response was increased five times by increasing the temperature from 7 to 19.5degreesC. The life span of the biosensor varied between I and 3 weeks after manufacturing.