Pathways for the degradation of 3,5-dimethyl-4-hydroxy-azobenzene-4'-sulfonic acid (I) and 3-methoxy-4-hydroxyazobenzene-4'-sulfonamide (II) by the manganese peroxidase and ligninase of Phanerochaete chrysosporium and by the peroxidase of Streptomyces chromofuscus have been proposed. Twelve metabolic products were found, and their mechanisms of formation were explained. Preliminary oxidative activation of the dyes resulted in the formation of cationic species, making the molecules vulnerable to the nucleophilic attack of water. Two types of hydrolytic cleavage were observed. Asymmetric splitting gave rise to quinone and diazene derivatives, while symmetric splitting resulted in the formation of quinone monoimine and nitroso derivatives. These unstable intermediates underwent further redox, oxidation, and hydrolytic transformation, eventually furnishing 11 organic products and ammonia.

Twenty-two azo dyes were used to study the influence of substituents on azo dye biodegradability and to explore the possibility of enhancing the biodegradabilities of azo dyes without affecting their properties as dyes by changing their chemical structures. Streptomyces spp. and Phanerochaete chrysosporium were used in the study. None of the actinomycetes (Streptomyces rochei A10, Streptomyces chromofuscus All, Streptomyces diastaticus A12, S. diastaticus A13, and S. rochei A14) degraded the commercially available Acid Yellow 9. * Corresponding author. t Publication no. 91512 of the Idaho Agricultural Experiment Station. enzyme back to its native state. Recently, we reported a new approach to enhancing aerobic azo dye transformation by P. chrysosponum and 3605

Five "4C-radiolabeled azo dyes and sulfanilic acid were synthesized and used to examine the relationship between dye substitution patterns and biodegradability (mineralization to C02) by a white-rot fungus and an actinomycete. 4-Amino-[U-14Cjbenzenesulfonic acid and 4-(3-sulfo-4-aminophenylazo)-[U-'4Clbenzenesulfonic acid were used as representative compounds having sulfo groups or both sulfo and azo groups. Such compounds are not known to be present in the biosphere as natural products. The introduction of lignin-like fragments into the molecules of 4-amino-[U-14Clbenzenesulfonic acid and 4-(3-sulfo-4-aminophenylazo)-[U-14C]benzenesulfonic acid by coupling reactions with guaiacol (2-methoxyphenol) resulted in the formation of the dyes 4-(3-methoxy-4-hydroxyphenylazo)-[U-14C]benzenesulfonic acid and 4-(2-sulfo-3'-methoxy-4'-hydroxy-azobenzene4-azo)-[U-14Clbenzenesulfonic acid, respectively. The synthesis of acid azo dyes 4-(2hydroxy-l-naphthylazo)-[U-14Cjbenzenesulfonic acid and 4-(4-hydroxy-l-naphthylazo)-[U-_4C]benzenesulfonic acid also allowed the abilities of these microorganisms to mineralize these commercially important compounds to be evaluated. Phanerochaete chrysosporium mineralized all of the sulfonated azo dyes, and the substitution pattern did not significantly influence the susceptibility of the dyes to degradation. In contrast, Streptomyces chromofuscus was unable to mineralize aromatics with sulfo groups and both sulfo and azo groups. However, it mediated the mineralization of modified dyes containing lignin-like substitution patterns. This work showed that lignocellulolytic fungi and bacteria can be used for the biodegradation of anionic azo dyes, which thus far have been considered among the xenobiotic compounds most resistant to biodegradation. Very specific structural changes in the azo dye molecules enhanced their biodegradability.

Actinomycete strains isolated from 2,4,6-trinitrotoluene (TNT)-contaminated and uncontaminated environments were compared for TNT tolerance and abilities to transform TNT. Regardless of previous TNT exposure history, no significant differences in TNT tolerance were seen among strains. Selected strains did not significantly mineralize [ 14 C]TNT. The actinomycetes did, however, transform TNT into reduced intermediates. The data indicate that, in actinomycete-rich aerobic environments like composts, actinomycetes will transform TNT into intermediates which are known to form recalcitrant polymers.

Eleven actinomycetes, isolated from the gut of worker termites (Macrotermes, Armitermes, Microcerotermes, Odontotermes), were identified as Streptomyces chromofuscus, S. chromogenus, S. diastaticus, and S. rochei. Their ability to grow on natural lignocellulosic substrates was tested in solid state fermentation experiments using wheat straw (C/N = 49.8) as a sole carbon source. Weight loss was 4.7–20.9% of the initial substrate, after 5 weeks at 30 °C; lignin and cellulose content decreased 2.0–16.1 and 3.5–32.9%, respectively. When the 11 Streptomyces were grown on wheat straw pretreated with (NH4)HCO3 (C/N = 28.2), weight loss was 9.3–29.9% of the initial substrate, indicating an overall enhancement of lignocellulose degradation. Weight, lignin, and cellulose losses were enhanced when S. chromofuscus (strain A2 and A11) and S. rochei A4 were grown on pretreated wheat straw instead of the untreated substrate. With S. rochei A10 the weight loss and lignin degradation were enhanced, while cellulolysis was slightly depressed. Weight loss and cellulose degradation were both enhanced when the remaining strains were grown on pretreated wheat straw. In this case, lignin degradation was depressed (S. chromofuscus A6 and A8, S. diastaticus A12, S. rochei A14) or remained essentially the same (S. diastaticus A3 and S. chromogenus A7). Key words: Streptomyces, wheat straw, degradation, lignin, cellulose.