prikaz prve stranice dokumenta Development of a fast, robust and sensitive spectrophotometric assay for measuring hydroxycinnamic acid decarboxylation reactions
  Download
PDF 1.35 MB
master's thesis
Development of a fast, robust and sensitive spectrophotometric assay for measuring hydroxycinnamic acid decarboxylation reactions
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2023. urn:nbn:hr:159:342678
Larva, Mirna
University of Zagreb
Faculty of Food Technology and Biotechnology
Department of Biochemical Engineering
Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology

Institutional repository: Repository of the Faculty of Food Technology and Biotechnology

Cite this document

Larva, M. (2023). Development of a fast, robust and sensitive spectrophotometric assay for measuring hydroxycinnamic acid decarboxylation reactions (Master's thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:342678

Larva, Mirna. "Development of a fast, robust and sensitive spectrophotometric assay for measuring hydroxycinnamic acid decarboxylation reactions." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2023. https://urn.nsk.hr/urn:nbn:hr:159:342678

Larva, Mirna. "Development of a fast, robust and sensitive spectrophotometric assay for measuring hydroxycinnamic acid decarboxylation reactions." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2023. https://urn.nsk.hr/urn:nbn:hr:159:342678

Larva, M. (2023). 'Development of a fast, robust and sensitive spectrophotometric assay for measuring hydroxycinnamic acid decarboxylation reactions', Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 04 August 2024, https://urn.nsk.hr/urn:nbn:hr:159:342678

Larva M. Development of a fast, robust and sensitive spectrophotometric assay for measuring hydroxycinnamic acid decarboxylation reactions [Master's thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2023 [cited 2024 August 04] Available at: https://urn.nsk.hr/urn:nbn:hr:159:342678

M. Larva, "Development of a fast, robust and sensitive spectrophotometric assay for measuring hydroxycinnamic acid decarboxylation reactions", Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2023. Available at: https://urn.nsk.hr/urn:nbn:hr:159:342678

Metadata
TitleDevelopment of a fast, robust and sensitive spectrophotometric assay for measuring hydroxycinnamic acid decarboxylation reactions
Title (croatian)Razvoj brze, robusne i osjetljive spektrofotometrijske metode za mjerenje reakcija dekarboksilacija hidroksicimetnih kiselina
AuthorMirna Larva
MentorAnita Slavica (mentor)
Committee memberRenata Teparic (predsjednik povjerenstva)
Committee memberBlaženka Kos (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Biochemical Engineering)
(Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology)
Zagreb
Defense date and country2023-05-11, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Biocatalytic decarboxylation of bio-based hydroxycinnamic acids yields p-hydroxystyrene derivates, which are important precursors of antioxidants, epoxy-coatings, adhesives and other polymeric materials. Bacillus subtilis phenolic acid decarboxylase (BsPAD) is a cofactor-independent enzyme that catalyses the cleavage of carbon dioxide from hydroxycinnamic acids such as p-coumaric, caffeic, ferulic and sinapic acid with high catalytic efficiency. Current methods to kinetically characterise the decarboxylation reactions rely on HPLC, MS, GC or NMR, which require extensive sample work-up while measuring only endpoint concentrations. In this work, we use a fast, robust and sensitive spectrophotometric assay that allows following decarboxylation reactions in real-time while avoiding product extraction. Absorption spectra of the pure reaction products were recorded after verifying the completeness of the reactions by TLC and NMR analyses. An optimized assay procedure was used to measure the kinetic constants (KM and Vmax values) of BsPAD for three substrates, which indicates the broad applicability of the assay.
Abstract (croatian)
Biokatalitičkom dekarboksilacijom prirodnih hidroksicimetnih kiselina dobiju se derivati p-hidroksistirena, koji su važni prekursori u proizvodnji antioksidansa, epoksi premaza, adheziva i drugih polimernih materijala. Dekarboksilaza fenolnih kiselina iz bakterije Bacillus subtilis (BsPAD) je enzim neovisan o kofaktoru, koji katalizira izdvajanje ugljičnog dioksida iz p-hidroksicimetnih kiselina poput p kumarinske, kofeinske, ferulinske i sinapinske kiseline uz visoku katalitičku učinkovitost. Metode koje se koriste za kinetičku karakterizaciju reakcija dekarboksilacije se temelje na korištenju HPLC, MS, GC ili NMR, i zahtijevaju opsežnu pripremu uzoraka, a njima se određuje samo krajnja koncentracija produkata ovih reakcija. U ovom diplomskom radu razvijena je brza, robusna i osjetljiva spektrofotometrijska metoda kojom se reakcija dekarboksilacije prati u realnom vremenu i kod koje nije potrebna ekstrakcija dobivenih produkata. Apsorpcijski spektri produkata snimljeni su nakon potvrde o kompletnoj dekarboksilaciji supstrata i to dvjema metodama - TLC i NMR. Optimizirana metoda korištena je za određivanje kinetičkih konstanti (KM i Vmax) za tri supstrata BsPAD, što ukazuje na široku primjenu ove razvijene metode.
Keywords
Keywords (croatian)
Languageenglish
URN:NBNurn:nbn:hr:159:342678
Study programmeTitle: Molecular Biotechnology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0) icon
Created on2024-07-09 10:07:32