Izolacija NADH oksidaze iz Lactobacillus sanfranciscencis

undergraduate thesis

• Title (english): Isolation of NADH oxidase from Lactobacillus sanfranciscencis
• Author: Elena Josić
• Mentor: Zvjezdana Findrik Blažević (mentor)
• Committee member: Zvjezdana Findrik Blažević (predsjednik povjerenstva)
• Committee member: Martina Sudar (član povjerenstva)
• Committee member: Marija Vuković Domanovac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2016-09-13, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering; Reaction Engineering

Abstract

Oksidaze koje ovise o prisustvu nikotinamid koenzima su od sve većeg interesa prilikom pripreme kiralnih molekula, bilo redukcijom prokiralnih prekursora ili iz oksidativne racemične smjese. Regeneracija oksidiranog i reduciranog oblika nikotinamid kofaktora je od iznimne važnosti jer je upotreba kofaktora u stehiometrijskim količinama izrazito skupa, a nemoguće ga je sintetizirati ili zamijeniti nekim jeftinijim produktom. U ovom radu opisane su metode regeneracije koenzima. Iako svi rezultati pokazuju da su trenutno razvijene metode pogodne za regeneraciju koenzima, većina njih zahtjeva daljnju optimizaciju kako bi proces postao održiv. U ovom radu je posebna pažnja posvećena NADH oksidazi iz Lactobacillus sanfranciscensis kao biokatalizatoru za regeneraciju koenzima. Karakterizacijom NADH oksidaze iz Lactobacillus sanfranciscensis ustanovljeno je enzim najaktivniji između pH 7.0 i 8.5, da producira vodu kao nusprodukt, a specifična aktivnost pročišćenog enzima iznosi 221 U/mg (pri pH 7.0). U radu će biti riječi o uzgoju mikroorganizma za njegovu izolaciju, metodama pročišćavanja, kao i o karakterizaciji enzima.

Abstract (english)

Oxidases which depend on nicotinamide coenzymes are of increasing interest for the preparation of chiral compounds, either by reduction of a prochiral precursor or by oxidative resolution of their racemate. The regeneration of oxidized and reduced nicotinamide cofactors is a crucial step because the use of these cofactors in stoichiometric amounts is too expensive for application and cannot be replaced by some cheaper compounds. This work describes the methods of cofactor regeneration. Although results indicate that those methods are suitable for the regeneration procedure, most of the processes need additional optimization to make them sustainable. In this work special attention is given to NADH oxidase from Lactobacillus sanfranciscensis as a biocatalyst for cofactor regenereration. The characterization of NADH oxidase from Lactobacillus sanfranciscensis has shown that the enzyme is the most active between pH 7.0 and 8.5. The specific activity of the purified enzyme is 221 U/mg (pH 7.0). This paper addresses the cultivation of the microorganism required for its isolation, enzyme purification methods, as well as the characterization of the enzyme.

• Keywords: NADH oksidaza; Lactobacillus sanfranciscensis
• Keywords (english): NADH oxidase; Lactobacillus sanfranciscensis
• Language: croatian
• URN:NBN: urn:nbn:hr:149:268345
• Study programme: Title: Environmental Engineering - Undergraduate study; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/ prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka ekoinženjerstva (sveučilišni/a prvostupnik/ prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka ekoinženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2018-02-05 11:07:38