A biosensor for NO3- containing immobilized dentrifying bacteria and a
reservoir of liquid growth medium for the bacteria was constructed. T
he bacteria did not have a N2O reductase and therefore reduced NO3- to
N2O, which was then subsequently quantified by a built-in electrochem
ical transducer for N2O, The only agents interfering with the determin
ation of NO3- were NO2- and N2O. The sensitivity for NO2- was identica
l to the one for NO3- whereas the sensitivity for N2O was 2.4 times hi
gher than for NO3-. Diffusive supply of electron donors to the bacteri
a from the built-in reservoir of growth medium ensured that the biosen
sor could work for 2-4 days. The tip diameter was down to 20 mu m, and
the sensors exhibited perfectly linear responses to nitrate in both f
reshwater and seawater. The detection limit was similar to 1 mu M. The
90% response time to changes in NO3- concentration was from 15 to 60
s at room temperature and about mice that at 6 degrees C, which was th
e lowest temperature for successful operation, The new NO3- biosensor
is a very useful tool for the study of nitrogen metabolism in nature.