De Lise, Federica (2018) Glycosidases and nanovesicles: novel biological tools for biotechnological applications. [Tesi di dottorato]

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Item Type: Tesi di dottorato
Lingua: English
Title: Glycosidases and nanovesicles: novel biological tools for biotechnological applications.
Creators:
CreatorsEmail
De Lise, Federicafederica.delise@unina.it
Date: 5 January 2018
Number of Pages: 172
Institution: Università degli Studi di Napoli Federico II
Department: dep03
Dottorato: phd012
Ciclo di dottorato: 30
Coordinatore del Corso di dottorato:
nomeemail
Sannia, Giovannigiovanni.sannia@unina.it
Tutor:
nomeemail
Di Donato, AlbertoUNSPECIFIED
Date: 5 January 2018
Number of Pages: 172
Uncontrolled Keywords: Glycosidases, OMVs, PP1Y, Purification
Settori scientifico-disciplinari del MIUR: Area 05 - Scienze biologiche > BIO/10 - Biochimica
Area 05 - Scienze biologiche > BIO/19 - Microbiologia generale
Date Deposited: 15 Jan 2018 09:50
Last Modified: 14 Mar 2019 11:54
URI: http://www.fedoa.unina.it/id/eprint/12271

Abstract

Biocatalysis represents a versatile and valuable tool for industrial biotechnologies. The use of enzymes as biocatalysts has reached its present industrial level, due to their optimal reaction selectivity, high reaction rates, high product purity, and a significant decrease in the generation of chemical waste. The use of enzymes in industrial applications has been limited by several factors, mainly the high cost of enzymes, their instability, and their availability in small amounts. To overcome these problems, the quest for new catalytic activities and the development of new technical approaches, such as enzyme immobilization, to improve their stability and practical applications, remain a central focus of the current biotechnological research. In this PhD thesis, the biotechnological potential of a novel bacterial glycosidase (-RHA), and the characterization of two possible immobilization systems, bacterial OMVs and eukaryotic EVs is reported. More in detail, an optimized expression and purification procedure allowed to characterize a novel -RHA from the microorganism Novosphingobium sp. PP1Y, which resulted to be appealing from a biotechnological point of view for its interesting catalytic behaviour. Moreover, mutagenesis experiments, allowed a preliminary identification of the amminoacidic residues responsible for the catalytic activity of rRHA-P, which could be further mutagenized to fine-tune rRHA-P catalytic efficiency on selected substrates. In addition, two different potential scaffolds from bacteria (OMVs) and from an eukaryotic cell line (EVs) were isolated and characterized. Both systems resulted to be appealing either for enzyme immobilization or for drug-delivery strategies. In particular, OMVs isolated from N. sp. PP1Y were characterized by a peculiar biochemical composition, which showed some differences with the originating whole cells. EVs isolated from human macrophages resulted to have differential effects on inflammation activation, and their potential as a valid alternative to bacterial OMVs for the development of novel delivery Biosystems is discussed.

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