Tyrosinase-based phenol remediation

Dissertation for the obtention of the Master Degree in Biotechnology

Nanobiotechnology is an emerging area full of potentialities to explore and reshape instituted knowledge. In this work, the main goal was to design and test biological remediation systems (a total of three) taking advantage of nanotechnology features, as a step towards future progress of sense-and-shoot combined systems. This task involved the development of active bionanoconjugates (BNC) with two different tyrosinases (Agaricus bisporus - AbT, and Tricoderma reesei - TrT) to be implemented in three suggested phenol remediation systems. AuNP-MUA (gold nanoparticles capped with11-mercapto-1-undecanoic acid) 1:5000 colloid was shown to be stable up to 100 mM ionic strength (sodium cloride) and able to form active BNCs with AbT. TrT:AuNP ratio for BNC formation is recommended as 100, supported by zeta potential and agarose gel mobility experimental data which, in turn, atone with the proposed theoretical estimation. The incubation of 100 ppm phenol with BNC-MUA-AbT for 60 minutes achieved 50% substrate conversion (the efficiency of the systems were also investigated using ultravioletvisible spectrophotometry (UV-Vis) complemented by standard addition method and solid-phase microextraction - gas chromatography (SPME-GC)).