Immobilisation of laccase on Eupergit supports and its application for the removal of endocrine disrupting chemicals in a packed-bed reactor

• Published in: Biodegradation (Dordrecht) Vol. 23; no. 3; pp. 373 - 386
• Main Authors: Lloret, L., Hollmann, F., Eibes, G., Feijoo, G., Moreira, M. T., Lema, J. M.
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01-06-2012; Springer; Springer Nature B.V
• Subjects: Adsorption; Animals; Aquatic Pollution; Biodegradation; Biodegradation of pollutants; Biological and medical sciences; Biomedical and Life Sciences; Bioreactors; Biotechnology; Chemicals; Color removal; Endocrine disruptors; Endocrine Disruptors - metabolism; Environment and pollution; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - metabolism; Estradiol; Ethinyl estradiol; Fundamental and applied biological sciences. Psychology; Fungal Proteins - chemistry; Fungal Proteins - metabolism; Geochemistry; Industrial applications and implications. Economical aspects; Kinetics; Laccase - chemistry; Laccase - metabolism; Life Sciences; Microbiology; Myceliophthora thermophila; Original Paper; Oxidases; Packed bed reactors; Polymers - chemistry; Reactors; Soil Science & Conservation; Sordariales - enzymology; Sorption; Temperature; Terrestrial Pollution; Waste Management/Waste Technology; Waste Water Technology; Water Management; Water Pollution Control; Eupergit; Estrone; 17β-estradiol; Covalent immobilisation; 17α-ethinylestradiol; Laccase; Biodegradation; Ethinylestradiol; Packed bed; Support; Enzyme; Immobilization; Endocrine disruptor; 17x-ethinylestradiol; Covalent bond; Oxidoreductases; Reactor; Application
• ISSN: 0923-9820; 1572-9729
• DOI: 10.1007/s10532-011-9516-7

Laccase from
Myceliophthora thermophila
was covalently immobilised on Eupergit C and Eupergit C 250L yielding specific activities of up to 17 and 80 U/g, respectively. Due to its superior activity, Eupergit C 250L was chosen for further research. The somewhat lower catalytic efficiency (based on the ratio between the turnover number and the Michaelis constant, k
cat
/K
M
) of the immobilised enzyme in comparison with that of the free enzyme was balanced by its increased stability and broader operational window related to temperature and pH. The feasibility of the immobilised laccase was tested by using a packed bed reactor (PBR) operating in consecutive cycles for the removal of Acid Green 27 dye as model substrate. High degrees of elimination were achieved (88, 79, 69 and 57% in 4 consecutive cycles), while the levels of adsorption on the support varied from 18 to 6%, proving that dye removal took place mainly due to the action of the enzyme. Finally, a continuous PBR with the solid biocatalyst was applied for the treatment of a solution containing the following endocrine disrupting chemicals: estrone (E1), 17β-estradiol (E2) and 17α-ethinylestradiol (EE2). At steady-state operation, E1 was degraded by 65% and E2 and EE2 were removed up to 80% and only limited adsorption of these compounds on the support, between 12 and 22%, was detected. In addition, a 79% decrease in estrogenic activity was detected in the effluent of the enzymatic reactor while only 14% was attained by inactivated laccase.