Enzyme immobilization on microfibrous or nanofibrous materials and their application in biotechnology
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Tato diplomová práce popisuje vývoj vhodných laboratorních postupů pro imobilizaci enzymů na nanovlákenné nosiče. Dále popisuje analytické metody pro rychlé a snadné stanovení jejich katalytické aktivity. Výzkum se zabývá různými technikami imobilizace a změnami parametrů procesu za účelem dosažení nejlepšího výsledku, kterým je kompromis mezi aktivitou a stabilitou imobilizovaného enzymu. Nejvyšší aktivity přes 150 U/g bylo dosaženo při kovalentním navázání lakázy z Trametes versicolor na nanovlákenné směsi polyamid 6/chitosan a polykaprolakton/silk fibroin. Nejvyšší aktivitu vykazovala lakáza imobilizovaná na polyamid 6 adsorpcí následovanou zesítěním. Vybrané vzorky byly použity pro degradaci dvou modelových endokrinních disruptorů (BPA a EE2). Tyto vzorky prokázaly výbornou katalytickou aktivitu během několika degradačních cyklů. Nanovlákna se osvědčila jako vhodný nosič pro imobilizaci enzymů s aplikací na čistění odpadních vod.
This diploma thesis describes a development of optimal laboratory techniques for enzyme immobilization on nanofibrous matrices and analytical methods for simple and fast determination of their enzymatic activity. The research deals with a screening of different immobilization procedures and adjusting parameters of the process to achieve the best result as a compromise between a high activity and satisfactory stability of the immobilized enzyme. The highest activities over 150 U/g were reached using polyamide 6/chitosan and polycaprolactone/silk fibroin blend nanofibers for covalent attachment while the operational stability showed laccase from Trametes versicolor immobilized on polyamide 6 nanofibers by adsorption followed by crosslinking. Selected samples were used for a degradation of two model endocrine disrupting chemicals (BPA and EE2). They showed excellent catalytic efficiency within several degradation cycles. Nanofibers appeared to be an optimal matrix for enzyme immobilization with application for wastewater treatment.
This diploma thesis describes a development of optimal laboratory techniques for enzyme immobilization on nanofibrous matrices and analytical methods for simple and fast determination of their enzymatic activity. The research deals with a screening of different immobilization procedures and adjusting parameters of the process to achieve the best result as a compromise between a high activity and satisfactory stability of the immobilized enzyme. The highest activities over 150 U/g were reached using polyamide 6/chitosan and polycaprolactone/silk fibroin blend nanofibers for covalent attachment while the operational stability showed laccase from Trametes versicolor immobilized on polyamide 6 nanofibers by adsorption followed by crosslinking. Selected samples were used for a degradation of two model endocrine disrupting chemicals (BPA and EE2). They showed excellent catalytic efficiency within several degradation cycles. Nanofibers appeared to be an optimal matrix for enzyme immobilization with application for wastewater treatment.
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lakáza, imobilizace, nanovlákna, chitosan, polyamid 6, silk fibroin, polykaprolakton, čištění odpadních vod, laccase, immobilization, nanofibers, chitosan, polyamide 6, silk fibroin, polycaprolactone, wastewater treatment