Investigations on the effects of pH, temperature, type and concentration of respiration substrates and oxygen tension on the reduction rate of derivatives of 1-phenylazo-2-naphthol and of a variety of textile dyes served as a basis for establishing a bioassay for strictly reproducible measurements of the microbial reduction rate of azo dyes. Standard organism was a strain of Bacillus cereus isolated from soil. Dye reduction occurred with the standard organism and other facultatively or obligatory aerobic bacteria in exclusively anoxic conditions. In principle, first order kinetics of decolorization were found. Reduction products may however inhibit the reaction. All dyes not measurably reduced by living cells of B. cereus were decolorized by cell extracts of the same species. Dyes adsorbed by the cell walls were in most cases reduced at slow rates and did not influence the simultaneous reduction of non-adsorbable dyes in the medium. The observations confirm the hypothesis advanced by Gingell and Walker (1971) of an intracellular, non-enzymatic reduction of azo compounds by reduced flavin nucleotides. The rate of permeation of the dyes through the cell membrane is the primordial ratelimiting step in the microbial decolorization of azo dyes. Sulfonic acid substitution seems to be an effective inhibitor of permeation.
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