Uklanjanje fenola i boja iz otpadne vode prirodnim i rekombinantnim oksidativnim enzimima
Removal of phenols and dyes from wastewater using native and recombinant oxidative enzymes
Author
Lončar, Nikola L.
Mentor
Vujčić, Zoran
Committee members
Vujčić, Miroslava
Gavrović-Jankulović, Marija

Metadata
Show full item recordAbstract
Boje i halogenovani fenoli koji imaju dezaktivirano aromatiĉno jezgro ĉine
znaĉajnu kategoriju veoma toksiĉnih i teško razgradljivih zagaĊivaĉa u raznim
industrijskim granama. Glavni cilj ove disertacije je bio dobivanje jeftinih rastvornih i
imobilizovanih enzima za uklanjanje fenola i boja iz otpadnih voda. Korišćeno je ĉetiri
prirodna (nativna) enzima (polifenoloksidaza iz krompira, tri lakaze na sporama
izolovanih sojeva Bacillus amyloliquefaciens, lakaza iz gljive Trametes versicolor,
kisele i bazne izoforme peroksidaze iz rena) i dva rekombinantna enzima
(hloroperoksidaza iz Caldariomyces fumago proizvedena u Aspergillus niger i lakaza iz
soja B. amyloliquefaciens 12B1 proizvedena u Escherichia coli).
Djelimiĉno preĉišćena polifenoloksidaza (PPO) iz krompira je imobilizovana na
razliĉitim nosaĉima. Od dobijenih biokatalizatora, tri sa najvećim aktivnostima PPO,
Eupergit C250L-PPO, Celit-PPO i CelulozaM-PPO, su testirani u reaktoru za uklanjanje
fenola, p-hlorfenola i p-bromfenola.... U sluĉaju 2,5 mM supstrata sa Eupergit C250LPPO,
postignuto je oko 45% razgradnje p-bromfenola, dok su p-hlorfenol i fenol
razgraĊeni 35% odnosno 20%. Testirana je i sposobnost višestruke upotrebe Eupergit
C250L-PPO imobilizata za uklanjanje p-hlorfenola. Iz eksperimenata sinteze novog
nosaĉa sa pipcima i njegove primjene za imobilizaciju PPO može se zakljuĉiti da je
imobilizovana PPO bila znatno otpornija na denaturaciju u odnosu na solubilni enzim.
Biokatalizator je testiran u šaržnom reaktoru za uklanjanje p-hlorfenola i p-bromfenola
iz vodenih rastvora. Postignuto je uklanjanje pomenutih fenola preko 90% pri
koncentraciji fenola 100 mg/L. Za oba halogenfenola TC-PPO je pokazao stepen
uklanjanja od preko 90% u prva tri ciklusa, nakon ĉega efikasnost opada do 60% nakon
šest ciklusa od po 8 ĉasova. Rastvornom PPO pod optimizovanim uslovima moguće je
ukloniti 93-99.9% boje nakon tretmana u trajanju od 1 ĉas sa 424-1700 U/mL PPO,
zavisno od boje. Pokazano je da je optimalno pH za proces obezbojavanja bilo 3,0.
Obezbojavanje je postignuto uz formiranje nerastvornih polimera koji su uklonjeni
filtrovanjem ili centrifugiranjem. Formiranje polimera potvrĊeno je infracrvenomspektroskopijom...
Dyes and phenols containing halogens which tend to deactivate the aromatic
nuclei constitute a significant category of highly toxic and difficult-to-degrade
pollutants from a wide variety of industries. Main goal of this dissertation was the
production of inexpensive soluble and immobilized enzymes for removal of phenols and
dyes from wastewater. Four native enzymes (potato polyphenoloxidase, three laccases
on spores of isolated strains of Bacillus amyloliquefaciens, laccase from Trametes
versicolor, acidic and basic isoforms of horseradish peroxidase) and two recombinant
enzymes (chloroperoxidase from Caldariomyces fumago produced in Aspergillus niger
and laccase from B. amyloliquefaciens 12B1 strain produced in Escherichia coli).
Partially purified potato polyphenol oxidase (PPO) was immobilized onto
different commercial and laboratory produced carriers. The three of the obtained
biocatalysts, with the highest PPO activities, namely Eupergit C250L-PPO, Celite-PPO
and CelluloseM-PPO, ...were tested in the batch reactor for phenol, p-chlorophenol and pbromophenol
removal. In the case of 2.5 mM substrates with Eupergit C250L-PPO,
around 45% removal of p-bromophenol was achieved, while p-chlorophenol and phenol
were removed 35% and 20%, respectively. The reusability of Eupergit C250L-PPO for
the removal of p-chlorophenol has been tested. From experiments of synthesis of new
tentacle carrier and its application for PPO immobilization we can conclude that
immobilized PPO was more resistant to denaturation when compared with its soluble
counterpart. Biocatalyst was tested in the batch reactor for p-chlorophenol and pbromophenol
removal from aqueous solution. More than 90% removal was achieved for
both halogenophenols at concentration of 100 mg/l from aqueous solution. For both
halogenophenols TC-PPO works with over 90% removal during first three cycles which
decrease to 60% removal efficiency after six cycles each of 8 hours duration. With
soluble PPO, under optimized conditions 93-99.9% removal of dyes was achieved after
1h using 424 – 1700 U/mL of PPO, depending on dye. Optimum pH for decolorization
process was found to be 3.0. Decolorization was accomplished via insoluble polymers
formations that were separated by filtration or centrifugation. Polymer formation was
confirmed with infrared spectroscopy...
Faculty:
Универзитет у Београду, Хемијски факултетDate:
06-12-2012Projects:
- Production, purification and characterization of enzymes and small molecules and their application as soluble or immobilized in food biotechnology, biofuels production and environmental protection (RS-172048)
- Interactions of natural products, their derivatives and coordination compounds with proteins and nucleic acids (RS-172055)