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master's thesis
Enzimska karakterizacija piruvat-kinaze bakterije Bacillus subtilis
Zagreb: University of Zagreb, Faculty of Science, 2012. urn:nbn:hr:217:787265
Kovač, Kristina
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Kovač, K. (2012). Enzimska karakterizacija piruvat-kinaze bakterije Bacillus subtilis (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:787265

Kovač, Kristina. "Enzimska karakterizacija piruvat-kinaze bakterije Bacillus subtilis." Master's thesis, University of Zagreb, Faculty of Science, 2012. https://urn.nsk.hr/urn:nbn:hr:217:787265

Kovač, Kristina. "Enzimska karakterizacija piruvat-kinaze bakterije Bacillus subtilis." Master's thesis, University of Zagreb, Faculty of Science, 2012. https://urn.nsk.hr/urn:nbn:hr:217:787265

Kovač, K. (2012). 'Enzimska karakterizacija piruvat-kinaze bakterije Bacillus subtilis', Master's thesis, University of Zagreb, Faculty of Science, accessed 27 December 2024, https://urn.nsk.hr/urn:nbn:hr:217:787265

Kovač K. Enzimska karakterizacija piruvat-kinaze bakterije Bacillus subtilis [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2012 [cited 2024 December 27] Available at: https://urn.nsk.hr/urn:nbn:hr:217:787265

K. Kovač, "Enzimska karakterizacija piruvat-kinaze bakterije Bacillus subtilis", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2012. Available at: https://urn.nsk.hr/urn:nbn:hr:217:787265

Metadata
TitleEnzimska karakterizacija piruvat-kinaze bakterije Bacillus subtilis
Title (english)Enzymatic characterization of Bacillus subtilis pyruvate kinase
AuthorKristina Kovač
MentorDamjan Franjević (mentor)
MentorIvan Mijaković (sumentor)
Committee memberDamjan Franjević (član povjerenstva)
Committee memberIvan Mijaković (član povjerenstva)
Committee memberZdravko Dolenec (član povjerenstva)
Committee memberBranka Pevalek-Kozlina (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2012, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Piruvat-kinaza je ključni enzim glikolize i metabolizma ugljika. Katalizira završni korak glikolitičkog puta, prenoseći fosforilnu skupinu fosfoenolpiruvata (PEP) na ADP, stvarajući ATP i piruvat. Reakcija je ireverzibilna, te predstavlja važno regulatorno mjesto u glikolizi. U ovom istraživanju piruvat-kinaza je izolirana iz aerobne Gram-pozitivne bakterije Bacillus subtillis, te pročišćena metodom afinitetne kromatografije do homogenosti. Enzimska rekacija se odvijala pri pH 7.5, uz prisustvo supstrata (PEP i ADP), te iona - K+ i Mg2+ koji s ADP-om tvori Mg2+-ADP kompleks, koji djeluje kao pravi substrat enzimu. Cilj istraživanja je bio utvrditi kinetičke parametre – Km i Vm piruvat-kinaze. Eksperimentalni podaci dobiveni spektrofotometrijskom metodom prikazani su Michealis-Menten modelom enzimske kinetike, te su pomoću linearizacijskih prikaza izračunate vrijednosti Km=1,2138 mM, Vm=227,062 μmol NADH/min/μg proteina za ADP, te Km=0,9081 mM, Vm=97,1125 μmol NADH/min/μg proteina za PEP. Navedeni su rezultati dobiveni Hanes-Woolf linearizacijskim prikazom.
Abstract (english)
Pyruvate kinase is key enzyme in glycolysis and carbon metabolim. It catalyzes the final step of glycolytic pathway, transferring phosphoryl group of phosphoenolpyruvate (PEP) to ADP, yielding ATP and pyruvate. Reaction is irreversible and it represents important regulatory point in glycolysis. In this research pyruvate kinase is isolated from aerobic Gram-positive bacteria Bacillus subtilis and purified to homogenity. Enzymatic reaction proceeded at pH 7.5 in presence of substrates (ADP and PEP) and ions - K+ and Mg2+. Mg2+ creates a Mg2+-ADP complex with ADP, which acts as a true substrate of the enzyme. Goal of this research was to identify kinetic parameters - Km i Vm of pyruvate kinase. Experimental data given by spectrofotometric method are shown with Michaelis-Menten model of enzyme kinetics. By means of linear plotting, values were calculated to Km=1,2138 mM, Vm=227,062 μmol NADH/min/μg protein for ADP and Km=0,9081 mM, Vm=97,1125 μmol NADH/min/μg protein for PEP. Mentioned results were given by Hanes-Woolf linear plot.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:787265
Study programmeTitle: Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra edukacije biologije (magistar/magistra edukacije biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
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Created on2018-04-18 11:34:27