De Luca, Viviana (2016) Heterologous expression and biochemical characterization of innovative bacterial carbonic anhydrases involved in the carbon cycle and microbial virulence. [Tesi di dottorato]

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Item Type: Tesi di dottorato
Lingua: English
Title: Heterologous expression and biochemical characterization of innovative bacterial carbonic anhydrases involved in the carbon cycle and microbial virulence.
Creators:
CreatorsEmail
De Luca, Vivianaviviana.deluca@unina.it
Date: 29 March 2016
Number of Pages: 114
Institution: Università degli Studi di Napoli Federico II
Department: Biologia
Scuola di dottorato: Scienze biologiche
Dottorato: Biologia avanzata
Ciclo di dottorato: 28
Coordinatore del Corso di dottorato:
nomeemail
Gaudio, Lucianogaudio@unina.it
Tutor:
nomeemail
Scudiero, RosariaUNSPECIFIED
Capasso, ClementeUNSPECIFIED
Date: 29 March 2016
Number of Pages: 114
Uncontrolled Keywords: carbonic anhydrase; carbon capture and storage; bioreactor; biomimetic capture; enzyme immobilization; pathogens; sulfonamides; anions; antiinfective; CA inhibitors; bacterial CAs
Settori scientifico-disciplinari del MIUR: Area 05 - Scienze biologiche > BIO/06 - Anatomia comparata e citologia
Area 05 - Scienze biologiche > BIO/10 - Biochimica
Date Deposited: 08 Apr 2016 09:25
Last Modified: 31 Oct 2016 09:22
URI: http://www.fedoa.unina.it/id/eprint/10767

Abstract

The conversion of carbon dioxide (CO2) to bicarbonate (HCO3-) and protons (H+) is a physiologically relevant reaction in all life kingdoms. The uncatalyzed hydration-dehydration reaction CO2 + H2O ⇋ HCO3- + H+ is slow at physiological pH and thus, in biological systems, the reaction is accelerated by enzymatic catalysts, called carbonic anhydrases (CAs, EC 4.2.1.1). CA isozymes have been found in virtually all mammalian tissues and cell types, where they function in CO2 transport and other physiological processes. Here, are described the biochemical characterization of thermostable and thermoactive CAs from extremophiles belonging to the genus Sulfurihydrogenibium. Moreover, it has been carried out a wide study concerning the inhibition profiles of CAs identified in the genome of pathogens causing disease in humans, such as Vibrio cholerae and Plasmodium falciparum. CAs identified in the genome of S. yellowstonense and S. azorense were found to be active at high temperatures and preserved its activity after immobilization on paramagnetic or polymeric supports. It has been demonstrated that these CAs are good candidates in the CO2 capture process. CAs from the pathogens aforementioned were biochemically characterized and an extensive inhibition profile was carried out using the classical CAs inhibitors such as sulfonamides and anions. These studies have contributed to the search of antibiotics with a novel mechanism of action. Additionally, the present thesis has contributed to the discovery of the η-CA, a new genetic families of CAs; to the introduction of a new technique, named protonography, useful for the identification of CA activity on a polyacrylamide gel; and to the phylogenetic analysis of the α-, β- and γ-CAs identified in the genome of the Gram-positive and Gram-negative bacteria.

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