Fourteen mono-ate dyes were used to study the effects of substitution patterns on the biodegradabilty of dimethyl-hydroxy-azobenzene 4'-sulfonic acids by Streptomyces chromofuscus A11. Two substitution patterns were analyzed: (i) all possible substitution patterns of the two methyl and hydroxy substitution groups, 2-hydroxy (3,5; 4,5; 5,6) dimethyl and 4-hydroxy (2,3; 2,5; 2,6; 3,5) dimethyl isomers of azobenzene 4'-sulfonic acid; and (ii) replacement of the sulfonic group with a carboxylic group in these sulfonated azo dyes, Tile structural pattern of the hydroxy group in para position relative to the azo linkage and of two methyl substitution groups in ortho position relative to the hydroxy group was the most susceptible to degradation, Replacement of the sulfonic group with a carboxylic group enhanced overall dye degradability by S. chromofuscus A11.