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undergraduate thesis
Izolacija i pročišćavanje halogenhidrin-dehalogenaze iz stanica genetski modificirane Escherichia coli
Zagreb: University of Zagreb, Faculty of Chemical Engineering and Technology, 2020. urn:nbn:hr:149:702999
Mihaljević, Iva
University of Zagreb
Faculty of Chemical Engineering and Technology

Institutional repository: Repository of Faculty of Chemical Engineering and Technology University of Zagreb

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Mihaljević, I. (2020). Izolacija i pročišćavanje halogenhidrin-dehalogenaze iz stanica genetski modificirane Escherichia coli (Undergraduate thesis). Zagreb: University of Zagreb, Faculty of Chemical Engineering and Technology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:149:702999

Mihaljević, Iva. "Izolacija i pročišćavanje halogenhidrin-dehalogenaze iz stanica genetski modificirane Escherichia coli." Undergraduate thesis, University of Zagreb, Faculty of Chemical Engineering and Technology, 2020. https://urn.nsk.hr/urn:nbn:hr:149:702999

Mihaljević, Iva. "Izolacija i pročišćavanje halogenhidrin-dehalogenaze iz stanica genetski modificirane Escherichia coli." Undergraduate thesis, University of Zagreb, Faculty of Chemical Engineering and Technology, 2020. https://urn.nsk.hr/urn:nbn:hr:149:702999

Mihaljević, I. (2020). 'Izolacija i pročišćavanje halogenhidrin-dehalogenaze iz stanica genetski modificirane Escherichia coli', Undergraduate thesis, University of Zagreb, Faculty of Chemical Engineering and Technology, accessed 09 July 2024, https://urn.nsk.hr/urn:nbn:hr:149:702999

Mihaljević I. Izolacija i pročišćavanje halogenhidrin-dehalogenaze iz stanica genetski modificirane Escherichia coli [Undergraduate thesis]. Zagreb: University of Zagreb, Faculty of Chemical Engineering and Technology; 2020 [cited 2024 July 09] Available at: https://urn.nsk.hr/urn:nbn:hr:149:702999

I. Mihaljević, "Izolacija i pročišćavanje halogenhidrin-dehalogenaze iz stanica genetski modificirane Escherichia coli", Undergraduate thesis, University of Zagreb, Faculty of Chemical Engineering and Technology, Zagreb, 2020. Available at: https://urn.nsk.hr/urn:nbn:hr:149:702999

Metadata
TitleIzolacija i pročišćavanje halogenhidrin-dehalogenaze iz stanica genetski modificirane Escherichia coli
Title (english)Isolation and purification of halohydrin dehalogenase from the genetically modified Escherichia coli
AuthorIva Mihaljević
MentorZvjezdana Findrik Blažević (mentor)
Committee memberZvjezdana Findrik Blažević (predsjednik povjerenstva)
Committee memberMartina Sudar (član povjerenstva)
Committee memberMarija Vuković Domanovac (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Chemical Engineering and Technology
Zagreb
Defense date and country2020-09-22, Croatia
Scientific / art field,
discipline and subdiscipline		TECHNICAL SCIENCES
Chemical Engineering
Abstract
U ovom radu provedena je izolacija i pročišćavanje enzima halogenhidrin-dehalogenaze (HheC). Najprije je uzgojena biomasa genetski modificiranih stanica Escherichia coli s visokim udjelom HheC. Nakon razbijanja bakterijskih stanica ultrazvukom, odvojeni su proteini od ostatka staničnog materijala centrifugiranjem te je dobiven sirovi enzimski ekstrakt. Kako bi se utvrdila uspješnost eksperimenta, mjerena je koncentracija proteina metodom po Bradfordu i aktivnost enzima HheC spektrofotometrijskim testom. Proces izolacije HheC je bitan za biokatalizu jer je otkriveno da ovaj enzim ima sposobnost kataliziranja reakcije otvaranja epoksida s malim “neprirodnim” anionskim nukleofilima, kao što su azid, cijanid, cijanat, tiocijanat, nitrit i formijat. Također, bitna primjena HHDH enzima je u proizvodnji optički čistih C3 ili C4 prekursora, na primjer, (R)-4-cijano-3-hidroksibutanoata za sintezu statina.
Abstract (english)
In this work, isolation and purification of the enzyme halohydrin dehalogenase (HheC) were carried out. Firstly, biomass of genetically modified Escherichia coli cells with a high HheC percentage was grown. After the disruption of bacterial cells by ultrasound, the proteins were separated from the rest of the cellular material by centrifugation and the crude enzyme extract was obtained. To determine the success of the experiment, the Bradford method for protein quantification and the spectrophotometric assay for HheC enzyme activity were used. The HheC isolation process is important for biocatalysis because it has been found that this enzyme has the ability to catalyze the epoxide ring-opening reaction with small "unnatural" anionic nucleophiles such as azide, cyanide, cyanate, thiocyanate, nitrite and formate. Also, the essential application of HHDH enzymes is in the production of optically pure C3 or C4 precursors, for example, (R)-4-cyano-3-hydroxybutanoate for statin synthesis.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:149:702999
ProjectNumber: IP-2018-01-4493
Title: Enzimska sinteza fluoriranih kiralnih građevnih blokova
Acronym: EnzyFluor
Leader: Maja Majerić-Elenkov
Jurisdiction: Croatia
Funder: HRZZ
Funding stream: IP
Study programmeTitle: Applied Chemistry - Undergraduate study; specializations in: Environmental chemistry and green technologies, Advanced materials and technologies, Applied organic chemistry
Course: Environmental chemistry and green technologies
Study programme type: university
Study level: undergraduate
Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) primijenjene kemije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) primijenjene kemije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2022-10-01
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Created on2023-05-05 12:04:58