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BIODEGRADATION OF DYES IN TEXTILE EFFLUENTS BY COVALENTLY CO-IMMOBILIZED MULTI-ENZYME OXIDASE SYSTEM TO SYNTHETIC

BIODEGRADATION OF DYES IN TEXTILE EFFLUENTS BY COVALENTLY CO-IMMOBILIZED MULTI-ENZYME OXIDASE SYSTEM TO SYNTHETIC POLYMER BEADS. L.Yotova, F.Tileva, R.Betcheva. INTRODUCTION. Dyes in textile effluents - role of dyeing (printing) technologies and dye structures

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BIODEGRADATION OF DYES IN TEXTILE EFFLUENTS BY COVALENTLY CO-IMMOBILIZED MULTI-ENZYME OXIDASE SYSTEM TO SYNTHETIC

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  1. BIODEGRADATION OF DYES IN TEXTILE EFFLUENTS BY COVALENTLY CO-IMMOBILIZED MULTI-ENZYME OXIDASE SYSTEM TO SYNTHETIC POLYMER BEADS. L.Yotova, F.Tileva, R.Betcheva

  2. INTRODUCTION • Dyes in textile effluents - • role of dyeing (printing) technologies and dye structures • Biotreatment of dyes in effluents – • classical aerobic sewage treatment process – adsorption on the sludge that • depends on the dye solubility • acid dyes – their high solubility leads to low adsorption ( depending on the degree of sulfonation) • direct dyes – high level of adsorption not depending on sulfonation • reactive dyes – very low degree of adsorption • basic dyes – high adsorption • disperse dyes – adsorption in the high to medium range • Reactive dyes • high consumption, applied in high concentrations in some technologies • relatively high concentrations in textile effluents • small molecules, high water solubility • decolourization • dearomatization

  3. Biodegradation of Sulfonated Azo Dyes (Goszczynski et all J. of Bacteriology, p.1339 -1347, 1994) Pathways for the aerobic microbial degradation of man-made sulfonated azo dyes by peroxidases from Phanaerochete chrisosporium Primarily asymmetric cleavage of azo linkage is supposed that leads to obtaining of sulfonamide and phenolic structures

  4. IMMOBILIZATION OF ENZYME SYSTEMS BY COVALENT BONDS • Advantages of covalently immobilized enzyme systems • immobilized whole cells keep a high level concentration of intracellular inducible multi-enzyme system • activity of these systems remains relatively high as far as the immobilization does not affect the cell integrity • possibility for use in reactors for continuous processes

  5. STUDY OBJECTIVE POSSUBILITIES OF REACTIVE DYE BIODEGRADATION BY COVALENTLY CO-IMMOBILIZED MULTI-ENZYME OXIDASE SYSTEM TO SYNTHETIC POLYMER BEADS SUBSTRATES INVESTIGATED phenol model dye - Phenol Red comercial dye - Bezactiv Brill.Red V-GG

  6. IMMOBILIZATION of the FUNGI STRAIN of Trichosporon cutaneum and Phanaerochete chrisosporium • carrier – activated acrylamide copolymer • treatment of granules with cell suspension • suspending in synthetic nutrient medium • ( 24 hours treatment) • washing • control for free cells ( spectrophotometrically) • microscopy

  7. Table 1

  8. PHENOL TRANSFORMATION ABILITY OF IMMOBILIZED CELLS • adaptation in a phenol medium (0,1g/l) • phenol transformation at 0,35 g/l , 0,65 g/l • and 1g/l substrate concentration • HPLC measurements of phenol concentration • (absorbance at 256 nm)

  9. KINETICS OF PHENOL CONVERTION Cells Trichosporon cutaneum 0,35 g/l phenol (Immobilized cells proliferate, remaining stable and active for 22 cycles) 0,65 g/l phenol 1,00 g/l phenol

  10. BIODEGRADATION OF PHENOL BY MULTYENZYME ENZYME PRODUCED IN IMMOBILZEDTrichosporon cutaneum CELLS Hydroxylation of phenolic structures Dearomatization by oxidase/hydrolase system in Trichosporon Cutaneum

  11. STUDY OF Phanarochete chrysosporium BEHAVIOUR • immobilized cells showed a relatively slow phenol degradation compared to Trichosporon cutaneum • source of exo-oxidase enzyme system containing lccase, lignin peroxidase and manganese peroxidase • ultrafiltration and immobilization of the above enzyme system • characterization of the immobilized system with respect of manganese peroxidase activity • Michaelis constant of the reaction with Phenol Red has the same range for free and immobilized manganese peroxidase

  12. Table 2 Phenol Red

  13. KINETICS OF BEZAKTIV Brill.RED V-GG DECOLOURIZATION BY THE IMMOBILIZED ENZYME SYSTEM PRODUCED OF Phanaerochete chrisosporium I step - dye water solution treatment (decolourization) II step – possibility for dearomatization with immobilized Trichosporon cutaneum cells should be additionally investigated

  14. CONCLUSIONS • Fungi can be immobilized at high concentration covalently to synthetic beads retaining a high level of stability and activity • Immobilized cells are viable and can be used as a source of multy-enzyme systems • Trichosporon cutaneum immobilized cells can be applied for phenolic structures dearomatization • Phanaerochete chrisosporiumoxidase multy-enzyme immobilized on synthetic beads decolourize monoazo dyes in water solution • Trichosporon cutaneum immobilized cells treatment of decolourized solutions have a potential to be used for dearomatization of above solutions

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