Rhodococcus erythropolis strain Y2, isolated from soil by enrichment culture using 1-chlorobutane, was able to utilize a range of halogenated aliphatic compounds as sole sources of carbon and energy. The ability to utilize 1-chlorobutane was conferred by a single halidohydrolase-type haloalkane dehalogenase. The presence of the single enzyme in cell-free extracts was demonstrated by activity stain polyacrylamide gel electrophoresis. The purified enzyme was a monomeric protein with a relative molecular mass of 34 kDa and demonstrated activity against a broad range of haloalkanes, haloalcohols and haloethers. The highest activity was found towards a, o disubstituted chloro-and bromo-C2-C6 alkanes and 4-chlorobutanol. The K,,, value of the enzyme for 1-chlorobutane was 0.26 mM. A comparison of the R. erythropolis Y2 haloalkane halidohydrolase with other haloalkane dehalogenases is discussed on the basis of biochemical properties and N-terminal amino acid sequence data.

Rhodococcus erythropolis Y2 produced two types of dehalogenase: a hydrolytic enzyme, that is an halidohydrolase, which was induced by C3 to C6 1-haloalkane substrates, and at least one oxygenase-type dehalogenase induced by C7 to C16 1-haloalkanes and n-alkanes. The oxygenase-type activity dehalogenated C4 to C18 1-chloroalkanes with an optimum activity towards 1-chlorotetradecane. The halidohydrolase catalysed the dehalogenation of a wide range of 1- and alpha,omega-disubstituted haloalkanes and alpha,omega-substituted haloalcohols. In resting cell suspensions of hexadecane-grown R. erythropolis Y2 the oxygenase-type dehalogenase had a specific activity of 12.9 mU (mg protein)-1 towards 1-chlorotetradecane (3.67 mU mg-1 towards 1-chlorobutane) whereas the halidohydrolase in 1-chlorobutane-grown batch cultures had a specific activity of 44 mU (mg protein)-1 towards 1-chlorobutane. The significance of the two dehalogenase systems in a single bacterial strain is discussed in terms of their contribution to the overall catabolic potential of the organism.

A microprocessor controlled flow injection system is described for the determination of vitamin C in foodstuffs. The system is based on amperometric detection at a wall-jet electrode coupled with an ascorbate oxidase packed bed. A commercially available Cartesian robotic auto-sampler-dilutor is used as a means of fully automating the sample handling and dilution. Dithiothreitol (DTT) is used to reduce dehydroascorbic acid to ascorbic acid and to stabilize ascorbic acid standard solutions. Initially, the system was connected in series with a high-performance liquid chromatography column and ultraviolet (UV) detector to allow identification of possible interferents and to allow comparative evaluation of results. The system showed a linear response to the concentration of L-ascorbic acid in the range 1-200 micrograms ml(-1) and was capable of detecting total vitamin C in a range of foodstuffs at a sample throughput of 15 samples h(-1). Correlations to existing methods of 0.98 were obtained.