1. Effect of green food processing technology on the enzyme activity in spelt flourMaja Leitgeb, Željko Knez, Gordana Hojnik Podrepšek, 2022, izvirni znanstveni članek Opis: In this research, a new approach to enzyme inactivation in flour was presented by supercritical
technology, considered a sustainable technology with lower energy consumption compared
to other technologies that use ultra-high temperature processing. Total protein concentration and
the activity of enzymes -amylase, lipase, peroxidase, polyphenol oxidase, and protease were determined
in flour pre-treated with scCO2. During the study, it was observed that the activity of enzymes
such as lipase and polyphenol oxidase, was significantly reduced under certain conditions of scCO2
treatment, while the enzymes -amylase and protease show better stability. In particular, polyphenol
oxidase was effectively inactivated below the 60% of preserved activity at 200 bar and 3 h, whereas
-amylase under the same conditions retained its activity. Additionally, the moisture content of the
scCO2-treated spelt flour was reduced by 5%, and the fat content was reduced by 58%, while the
quality of scCO2-treated flour was maintained. In this regard, the sustainable scCO2 process could be
a valuable tool for controlling the enzymatic activity of spelt flour since the use of scCO2 technology
has a positive effect on the quality of flour, which was verified by the baking performance of spelt
flour with the baked spelt bread as an indicator of quality. Ključne besede: food treatment, spelt flour, proteins, supercritical carbon dioxide, enzyme inactivation, defatted flour, carbon dioxide solubility Objavljeno v DKUM: 17.04.2025; Ogledov: 0; Prenosov: 1
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2. Influence of supercritical carbon dioxide on the activity and conformational changes of α-amylase, lipase, and peroxidase in the solid state using white wheat flour as an exampleMilena Ivanović, Željko Knez, Maja Leitgeb, 2023, izvirni znanstveni članek Ključne besede: enzyme inactivation, supercritical carbon dioxide, a-amilaze, lipase, antioxidants, horseradish peroxidase, structural changes, CD spectroscopy Objavljeno v DKUM: 22.04.2024; Ogledov: 183; Prenosov: 15
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3. Use of non-conventional cell disruption method for extraction of proteins from black yeastsMaja Čolnik, Mateja Primožič, Željko Knez, Maja Leitgeb, 2016, izvirni znanstveni članek Opis: The influence of pressure and treatment time on cells disruption of different black yeasts and on activities of extracted proteins using supercritical carbon dioxide process was studied. The cells of three different black yeasts Phaeotheca triangularis, Trimatostroma salinum, and Wallemia ichthyophaga were exposed to supercritical carbon dioxide (SC $CO_2$) by varying pressure at fixed temperature (35°C). The black yeasts cell walls were disrupted, and the content of the cells was spilled into the liquid medium. The impact of SC $CO_2$ conditions on secretion of enzymes and proteins from black yeast cells suspension was studied. The residual activity of the enzymes cellulase, $beta$-glucosidase, $alpha$-amylase, and protease was studied by enzymatic assay. The viability of black yeast cells was determined by measuring the optical density of the cell suspension at 600 nm. The total protein concentration in the suspension was determined on UV–Vis spectrophotometer at 595 nm. The release of intracellular and extracellular products from black yeast cells was achieved. Also, the observation by an environmental scanning electron microscopy shows major morphological changes with SC $CO_2$-treated cells. The advantages of the proposed method are in a simple use, which is also possible for heat-sensitive materials on one hand and on the other hand integration of the extraction of enzymes and their use in biocatalytical reactions. Ključne besede: P. triangularis, W. ichtyophaga, T. salinum, supercritical carbon dioxide, enzyme activity, cells Objavljeno v DKUM: 10.07.2017; Ogledov: 1546; Prenosov: 406
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4. Phase equilibrium measurements and data correlation for the ternary system oleic acid + 1-octanol + carbon dioxideChiara Giulia Laudani, Mateja Primožič, Željko Knez, Maja Leitgeb, 2009, izvirni znanstveni članek Opis: A study of the high-pressure phase equilibria in the ternary system oleic acid/1- octanol/supercritical CO2 was performed to obtain information for optimization of enzymatic synthesis of n-octyl oleate. Equilibrium data were measured at temperatures of 308.15, 323.15 and 343.15 K over a pressure range from 1 to 25 MPa. Two different apparatuses: a Variable Volume View Cell and a Batch Stirred Tank were used employing synthetic and static-analytic measurement methods, respectively. The measured solubility of dense CO2 in the binary liquid mixture oleic acid/1-octanol covered the range from 0.0781 to 0.7686 of CO2 molar fraction. CO2 solubility increased with increasing pressure up to 10 MPa. At higher pressures, no significant increase was observed. The Soave-Redlich-Kwong equation-of-state (SRK-EoS) with quadratic mixing rules was successfully used for data correlation in the whole P-T-x region studied. Ključne besede: data correlation, 1-octanol, oleic acid, supercritical carbon dioxide, Soave-Redlich-Kwong equation-of-state, vapor-liquid equilibria Objavljeno v DKUM: 05.07.2017; Ogledov: 1558; Prenosov: 120
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5. Enzymatic esterification of glycerol and stearic acid in non-conventional mediaZsófia Csanádi, Katalin Bélafi-Bakó, Eszter Szentgyörgyi, László Gubicza, Željko Knez, Maja Leitgeb, 2010, izvirni znanstveni članek Opis: Ionic liquids as trihexyl-tetradecyl-phophonium-dicyanamide (Cyphos 105) and cocosalkyl-pentaethoxi-methyl-ammonium-methosulfate (Ammoeng 100) were applied for the esterification of stearic acid and glycerol using Candida antarctica lipase (Novozyme 435). When only ILs were applied as solvents at 1:15 initial substrate molar ratio the conversion was 76 and 78 % in the case of two kinds of ILs, respectively. Mixed the ILs and supercritical $CO_2$ the conversion reached 79 and 86 %. The conversion was found highest in supercritical $CO_2$, reached 90 %. Moreover formation of glycerol-di-stearate is much lower in the case of ILs comparing with $SCCO_2$. Ključne besede: glycerol-mono-stearate, Lipase, ionic liquids, supercritical $CO_2$, carbon dioxide Objavljeno v DKUM: 10.07.2015; Ogledov: 1488; Prenosov: 203
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7. HIGH PRESSURE PHASE EQUILIBRIUM OF THE BINARY SYSTEM TWEEN 80 AND CARBON DIOXIDENatalija Novak, 2014, diplomsko delo Opis: In the frame of the diploma thesis we researched the phase equilibrium of the binary system Tween 80 and CO2. Equilibrium compositions of the system have been determined with a static analytical method at temperature 40, 60 and 80 °C and pressures up to 300 bar. The obtained data was plotted in isothermal diagrams where mass percentage is presented as a function of pressure.
In this work also the viscosity of Tween 80 was determined in the temperature range between 20 and 80°C and shear rates up to 180 s-1. The measurements were obtained by using coaxial cylinder and compared with available literature data. Ključne besede: supercritical fluids, phase equilibrium, carbon dioxide, Tween 80 Objavljeno v DKUM: 22.04.2014; Ogledov: 2035; Prenosov: 93
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8. Thermodynamic properties of coenzyme Q10 in supercritical carbon dioxideDarja Pečar, Valter Doleček, 2007, izvirni znanstveni članek Opis: The densities of coenzyme Q10 in supercritical carbon dioxide were measured at 308.15, 313.15, 323.15 and 333.15 K and in the pressure range from 10 to 40 MPa with the vibrating tube densimeter. The reliability of the technique has been verified in our previous works and in the present work it was checked comparing our experimental densities of pure carbon dioxide with literature data. The partial molar volumes, isothermal compressibilities, isobaric thermal expansivities, internal pressures and cluster sizes were calculated from obtained densities. Ključne besede: koenzim Q10, density measurements, partial molar volume, isothermal compressibility, isobaric thermal expansivity, internal pressure, supercritical carbon dioxide Objavljeno v DKUM: 01.06.2012; Ogledov: 1927; Prenosov: 148
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9. Chemical composition of Juniperus communis L. fruits supercritical CO2 extracts: dependence on pressure and extraction timeBranislava Barjaktarović, Milan Sovilj, Željko Knez, 2005, izvirni znanstveni članek Opis: Ground fruits of the common juniper (Juniperus communis L), with a particle size range from 0.2500.400 mm, forming a bed of around 20.00 +/- 0.05 g, were extracted with supercritical CO2 at pressures of 80,90, and 100 bars and at a temperature of 40 De. The total amount of extractable substances or global yield (mass of extract/mass of raw material) for the supercritical fluid extraction process varled from 0.65 to 4.00"10 (wt). At each Investigated pressure, supercriticaI CO2 extract fractions collected In successive time intervals over the course of the extraction were analyzed by capillary gas chromatography, using flame ionization (GO-FIO) and mass spectrometric detection (GC-MS). More than 200 constituents were detected In the extracts, and the contents of 50 compounds were reported in the work. Dependence of the percentage yields of monoterpene, sesquiterpene, oxygenated monoterpene, and oxygenated sesquiterpene hydrocarbon groups on the extraction time was investigated, and conditions that favored the yielding of each terpene groups were emphasized. At all pressures, monoterpene hydrocarbons were almost completely extracted from the berries in the first 0.6 h. It was possible to extract oxygenated monoterpenes at 100 bar in 0.5 h and at 90 bar in 1.2 h. Contrary to that, during an extraction period of 4 h at 80 bar, it was possible to extract only 75% of the maximum yielded value of oxygenated monoterpene at 100 bar. Intensive extraction of sesquiterpenes could be by no means avoided at any pressure, but at the beginning of the process (the first 0.5 h) at 80 bar, they were extracted about a and 3 times slower than at 100 and 90 bar, respectively. Oxygenated sesquiterpenes were yielded at fast, constant extraction rates at 100 and 90 bar In 1.2 and 3 h, respectively. This initial fast extraction period was consequently foIlowed by much slower extraction of oxygenated sesquiterpenes. Ključne besede: chemical processing, high pressure technology, CO2, supercritical fluid extraction, pressure, extraction time, Juniperus communis, oxygenated terpenes, juniper berry oil, carbon dioxide Objavljeno v DKUM: 01.06.2012; Ogledov: 2229; Prenosov: 59
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10. Lipase-catalyzed transesterification of (R,S)-1-phenylethanol in SC $CO_2$ and in SC $CO_2$/ionic liquid systemsMuzafera Paljevac, Željko Knez, Maja Leitgeb, 2009, izvirni znanstveni članek Opis: Commercial immobilized lipase B from Candida antarctica (CALB) was successfully applied to catalyzing the transesterification of (R,S)-1-phenylethanol in supercritical carbon dioxide and in supercritical carbon dioxide/ionic liquid biphasic system. Firstly, the variables affecting the performance of CALB in transesterification reactions in supercritical carbon dioxide, such as CALB concentration, temperature and pressure, were studied. An increase in the conversion and in the reaction rate was observed as the CALB/substrate ratio, temperature and pressure, were increased from 4.3 to 19.9, from 40 °C to 80 °C and from 8 MPa to 10 MPa, respectively. Further increase in temperature from 80 °C to 120 °C and pressure from 10 MPa to 30 MPa resulted in lower conversion and lower initial reaction rate. Furthermore, different vinyl esters were used as acyl donors for CALB-catalyzed transesterification of (R,S)-1-phenylethanol in supercritical carbon dioxide. The highest initial reaction rate was attained with vinyl butyrate, although 50% conversion was attained faster when vinyl acetate was used as acyl donor.
Secondly, in transesterification of (R,S)-1-phenylethanol, performed in supercritical carbon dioxide/ionic liquid biphasic system, influence of concentration of 1-butyl-3-methylimidazolium tetrafluoroborate [bmim][BF$_4$] was studied. The addition of 50 mmol (70% w/w reaction mixture) of [bmim][BF$_4$] to the reaction system gave the best result in terms of transesterification rate. Ključne besede: candida antarctica lipase B, transesterification, (R, S)-1-phenylethanol, ionic liquids, vinyl acetate, supercritical carbon dioxide Objavljeno v DKUM: 31.05.2012; Ogledov: 2480; Prenosov: 241
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