Protein kinase ASK1 (apoptosis signal-regulating kinase 1) is a member of the mitogen- activated protein kinase kinase kinase (MAP3K) family and plays a crucial role in immune and stress responses. Since the increased activity of ASK1 has been linked to the development of several diseases including cancer, cardiovascular and neurodegenerative diseases, this enzyme is a promising target for therapeutical intervention in these pathologies. The molecule of ASK1 consists of 1374 amino acid residues, but catalytic activity possesses only a kinase domain located approximately in the middle of the molecule. The activity of ASK1 is regulated by interactions with various proteins including the 14-3-3 protein. This protein recognizes a phosphorylated motif around Ser966 at the C-terminus of the catalytic domain of ASK1. This binding interaction inhibits ASK1 through unknown mechanism. ASK1 under stress conditions, such as oxidative stress, is dephosphorylated at Ser966 and the 14-3-3 protein dissociates. This dissociation is then one of the factors that lead to the activation of ASK1. The aim of this diploma thesis was to prepare a complex of the catalytic domain of ASK1 with the 14-3-3 protein for subsequent structural studies. Both proteins were expressed in E. coli cells and successfully purified. In...

Naudojant chronopotenciometrijos metodą buvo tirta sidabro elektronusodinimo iš sulfitinių tirpalų kinetika. Eksperimentiniai tyrimai patvirtino negausius žinomus literatūrinius duomenis, kad sidabro elektrokristalizacija iš sulfitinių elektrolitų yra apsprendžiama lėta sidabro ad-atomų kristalizacija galvaninės dangos paviršiuje. Tai ypač akivaizdu pusiausvyrinio potencialo srityje. Nustatyta, kad patikimiausi rezultatai, tiriant krūvio pernešimo stadiją, gaunami maksimaliai nutolus nuo pusiausvyros potencialo. Tai lengviausia realizuoti chronopotenciometrijos sąlygomis, o nustatytas mainų srovės tankis svyruoja tarp 0,9 (kai tirpalo sudėtis 0,003 ir 0,115 ) iki 2,46 (kai tirpalo sudėtis 0,1 ir 0,77 ). Patikimiausi rezultatai buvo gauti matuojant mainų srovės tankių priklausomybes nuo laisvų sulfito jonų koncentracijos izopotencialių ( 0,300 ) tirpalų serijoje. Visuose izopotencialiuose tirpaluose buvo nustatyti krūvio pernašos viršįtampiai -60 – -160 intervale. Naudojantis lygtimi ir mainų srovės tankio priklausomybės nuo laisvų sulfito jonų koncentracijos grafiku iš eksperimentinių duomenų nuolinkio kampo apskaičiuotas reakcijos laipsnis 0,67. Atsižvelgiant į galimas paklaidas, tikėtina, kad jis yra lygus 1. Gauti rezultatai rodo, kad artimas vienetui reakcijos laipsnis sulfito jonų atžvilgiu atitinka teiginį, jog betarpiškai fazių sąlyčio riboje redukuojasi kompleksinė dalelė . Atsiradusios paklaidos greičiausiai yra susiję su tirpalo komponentų... [toliau žr. visą tekstą] / Silver electrodeposition from sulfite electrolytes is considered to be one of the most promising processes to replace the extremely toxic traditional cyanide-based silver plating solutions with the additional advantage, that sulfite electrolytes can be successfully applied for silver deposition for micro-electro-mechanical systems (MEMSs). Mechanism of metal complexes electroreduction is an important result, especially when it may have a wide range of applications in the future. However, electrodeposition also has several shortcomings, including sulfite ion oxidation by oxygen in air and relatively negligible cathodic current densities, which are too low for qualitative galvanotechnique. There are very few works dedicated to the chemistry and electrochemistry of silver sulfite complexes. Most publications related with this subject are concerned with the questions of applicability. The main objective of this study was the investigation of the silver sulfite complexes electroreduction kinetics. To achieve this goal, we studied the dependence of the exchange current density on the free sulfite ions concentration in a series of isopotential solutions. Our studies also allowed to determine the mechanism of this electrochemical charge transfer reaction.

Silver electrodeposition from sulfite electrolytes is considered to be one of the most promising processes to replace the extremely toxic traditional cyanide-based silver plating solutions with the additional advantage, that sulfite electrolytes can be successfully applied for silver deposition for micro-electro-mechanical systems (MEMSs). Mechanism of metal complexes electroreduction is an important result, especially when it may have a wide range of applications in the future. However, electrodeposition also has several shortcomings, including sulfite ion oxidation by oxygen in air and relatively negligible cathodic current densities, which are too low for qualitative galvanotechnique. There are very few works dedicated to the chemistry and electrochemistry of silver sulfite complexes. Most publications related with this subject are concerned with the questions of applicability. The main objective of this study was the investigation of the silver sulfite complexes electroreduction kinetics. To achieve this goal, we studied the dependence of the exchange current density on the free sulfite ions concentration in a series of isopotential solutions. Our studies also allowed to determine the mechanism of this electrochemical charge transfer reaction. / Naudojant chronopotenciometrijos metodą buvo tirta sidabro elektronusodinimo iš sulfitinių tirpalų kinetika. Eksperimentiniai tyrimai patvirtino negausius žinomus literatūrinius duomenis, kad sidabro elektrokristalizacija iš sulfitinių elektrolitų yra apsprendžiama lėta sidabro ad-atomų kristalizacija galvaninės dangos paviršiuje. Tai ypač akivaizdu pusiausvyrinio potencialo srityje. Nustatyta, kad patikimiausi rezultatai, tiriant krūvio pernešimo stadiją, gaunami maksimaliai nutolus nuo pusiausvyros potencialo. Tai lengviausia realizuoti chronopotenciometrijos sąlygomis, o nustatytas mainų srovės tankis svyruoja tarp 0,9 (kai tirpalo sudėtis 0,003 ir 0,115 ) iki 2,46 (kai tirpalo sudėtis 0,1 ir 0,77 ). Patikimiausi rezultatai buvo gauti matuojant mainų srovės tankių priklausomybes nuo laisvų sulfito jonų koncentracijos izopotencialių ( 0,300 ) tirpalų serijoje. Visuose izopotencialiuose tirpaluose buvo nustatyti krūvio pernašos viršįtampiai -60 – -160 intervale. Naudojantis lygtimi ir mainų srovės tankio priklausomybės nuo laisvų sulfito jonų koncentracijos grafiku iš eksperimentinių duomenų nuolinkio kampo apskaičiuotas reakcijos laipsnis 0,67. Atsižvelgiant į galimas paklaidas, tikėtina, kad jis yra lygus 1. Gauti rezultatai rodo, kad artimas vienetui reakcijos laipsnis sulfito jonų atžvilgiu atitinka teiginį, jog betarpiškai fazių sąlyčio riboje redukuojasi kompleksinė dalelė . Atsiradusios paklaidos greičiausiai yra susiję su tirpalo komponentų... [toliau žr. visą tekstą]

Water pollution by organic contaminants and other anthropogenic substances is one of the major global problems nowadays. The aquatic contaminants include pesticides, pharmaceuticals and personal care products (PPCPs), steroid hormones and synthetic dyes. Traditional processes such as UV irradiation at disinfection doses, coagulation, flocculation, precipitation, microfiltration and ultrafiltration are ineffective for the complete removal of organic contaminants in water, whereas the so called advanced oxidation processes (AOPs) are very effective in the oxidation of numerous organic compounds. The most widely used AOPs include heterogeneous photocatalysis based on near UV or solar visible irradiation, electrolysis, the Fenton's reaction, ozonation, ultrasound and wet air oxidation. AOPs are based on the generation of highly reactive hydroxyl radicals (oOH), which will attack the organic pollutants. However, the lifetimes of these radicals are so short to utilize them effectively. Therefore, the direct radical generations by non-thermal plasmas are widely researched. Three different types of plasma discharges (Dielectric Barrier Discharge, DBD, Gliding Arc Discharge, GAD, and Electrohydraulic Spark Discharge, ESD) were applied to investigate the degradation kinetics of the target pollutants in water. Firstly, the degradation of atrazine, verapamil and hydrocortisone was conducted successfully in the DBD reactor. After 90 min DBD treatment all the target pollutants were almost completely removed. Presence of the intermediates was confirmed by HPLC/UV analysis. The efficiency of dielectric barrier discharge on the degradation of atrazine and hydrocortisone was investigated. Atrazine was almost completely degraded after 90 min DBD treatment. The rate constant of this process was 0.029 min-1 and the corresponding half-life time was 24 min. After 90 min treatment time 54% of the atrazine was converted to CO2. Hydrocortisone was also completely degraded after 90 min DBD treatment. The rate constant of this process was 0.050 min-1 and the corresponding half-life time was 14 min. After five hours of treatment 21% of the hydrocortisone was converted to CO2. Structure of the intermediates was identified by HPLC/MS analysis. Plausible mechanisms for the degradation of target pollutants were also proposed. Degradation kinetics and degradation mechanism of verapamil in water under GAD was investigated. In the final part, the degradation kinetics of verapamil and atrazine by using electrohydraulic spark discharge and ozonation was investigated. During spark discharge, 87% of verapamil and 83% of atrazine were degraded within 40 min. The rate constants of the degradation processes were 0.065 min-1 and 0.094 min-1 for verapamil and atrazine respectively. The power introduced in the discharge was 60 W and is very low compared to the power of GAD. As a result, relatively high energy yields 5.1 × 10-2 g/kWh and 2.3 × 10-2 g/kWh were achieved for verapamil and atrazine respectively. Verapamil was completely removed by ozonation within 1.5 min., while atrazine was almost completely removed within 4 min. The rate constants of the degradation processes were 2.56 min-1 and 0.769 min-1 for verapamil and atrazine respectively. The power applied during ozonation was very low (20 W), compared to plasma discharges, leading to high energy yields 9.4 g/kWh and 1.6 g/kWh for verapamil and atrazine respectively. The degradation kinetics of verapamil and atrazine in water under DBD, GAD, ESD and ozonation were compared. The shortest half-lives of verapamil and atrazine were observed during ozonation. So, we can say that among our AOP systems, ozonation is the most efficient in degrading verapamil and atrazine in water.

In an effort to increase the thermodynamic stability and the kinetic inertness of the complexes, the five new azamacrobicyclic ligands derived from TACN, cyclen, and cyclam have been prepared. The ligands were decorated with phosphinate or phosphonate pendant arms to maintain fast complexation. Since the ascending importance of targeted diagnostic and therapy, the bone-targeted non-bridged cyclam derivative with phosphinate-bis(phosphonate) pendant arm (H5TE1PBP) has also been synthesized. The ligands were studied with respect to their application. The bridged TACN (H2bpbtacn) and cyclen (H4bpbcen) ligands show high macrocyclic basicity (logK1 = 12.25 and 12.70, respectively). The thermodynamic stability of H2bpbtacn with Cu(II) ion is more than ten orders of magnitude lower than that of the NOTA ligand. The stability constants of H4bpbcen with Cu(II) and Zn(II) ions are comparable to those given for the DOTA. The stability of Ln(III)-bpbcen complexes is 7-10 orders of magnitude lower compared to DOTA complexes. For both ligands, the lower thermodynamic stability of the complexes is attributed to the high rigidity of the ligand structure. The bridged cyclam derivatives with phosphonate (H4TE2P), bis(phosphinate) (H4TE2bpin), or phosphinate (H2TE2PH) pendants are characterized by high stability of...

This thesis describes research that has been carried out during the years 2009-2013 as a part of my PhD project related to the method of selected ion flow tube mass spectrometry (SIFT-MS) and its application in interdisciplinary areas of research. SIFT-MS is a method that allows accurate quantification of trace gases and vapours presented in humid air with the focus on human breath; without any sample preparation and in real time. The thesis is divided into several parts. The first part reviews the history of mass spectrometry as a background for the quantitative analytical methods as PTR-MS and SIFT-MS. The second part discusses the detailed history of development of SIFT-MS, starting from principles of selected ion flow tube (SIFT) technique that has been used for study of ion-molecule reactions in the gas phase and forms the basis of SIFT-MS. The next part discusses volatile organic compounds of different biological origin: bacterial, plant and human breath metabolites that can be analyzed in real time using SIFT-MS. The main part "Results and Discussion" is divided into several subsections that serve as commentaries to the enclosed research papers published in peer reviewed journals. The first is a detailed step by step overview of the kinetics of ion molecule reactions which is the basis of...

Slow complexation is one of the major limitations of current macrocyclic chela- tors utilized in nuclear medicine for complexation of metal radionuclides. This property can be improved by ligand design. Among metal radioisotopes, the copper ones (e.g. 60Cu, 61Cu, 62Cu, 64Cu and 67Cu) have become commonly available in recent years and cyclam-derived ligands are the most suitable ligands for Cu2+ complexation. To alter the complexation rate, bis(phosphinic acid) group is promising unit as it is able to complex metal ions in acidic solu- tions. However, geminal bis(phosphinates) represent poorly studied group of compounds and, thus, it is challenging to uncover their properties. Several simple amino-bis(phosphinates) were synthesized and its acid-base and coordi- nation properties were studied. Unlike structurally similar geminal bis(phosphonates), the title compounds showed negligible adsorption onto hydroxyapatite (commonly used model of bone tissue). The obtained knowledge dealing with geminal bis(phosphinates) was utilized in synthe- sis of two novel cyclam derivatives. The ligands (bearing either one geminal bis(phosphinate) or one geminal phosphino-phosphonate pendant arm) were synthesized by highly efficient procedure. The corresponding Cu2+ complexes are formed with very high rates and show...

<p>Hemijskim transformacijama se iz biljnih ulja dobijaju vredni &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; derivati, poput epoksidovanih biljnih ulja, koji se<br />koriste u hemijskoj i polimernoj industriji. Kvalitet, pa<br />time i primena epoksidovanih biljnih ulja, zavise od<br />sadržaja epoksidnih grupa u derivatizovanom ulju, koji bi<br />trebalo da je &scaron;to veći. Kako epoksidne grupe nastaju<br />oksidacijom dvostrukih veza u trigliceridima kao<br />dominantnoj grupi jedinjenja u biljnim uljima, pogodna<br />sirovina za epoksidovanje su visoko nezasićena ulja,<br />kakvo je laneno.<br />Proizvodnja epoksidovanih biljnih ulja zahteva izbor<br />takvih procesnih uslova pri kojima bi se postigli &scaron;to<br />potpunija konverzija dvostrukih veza i &scaron;to veća<br />selektivnost procesa u odnosu na epoksidnu grupu. Zato je<br />kao cilj ove doktorske disertacije postavljeno određivanje<br />optimalnih vrednosti procesnih uslova epoksidovanja<br />lanenog ulja persirćetnom kiselinom formiranom in situ iz<br />sirćetne kiseline i 30% vodenog rastvora vodonik<br />peroksida u prisustvu jonoizmenjivačke smole kao<br />katalizatora. Određivanje je izvedeno primenom<br />metodologije odzivne povr&scaron;ine, kao i kori&scaron;ćenjem u ovoj<br />disertaciji predloženih kinetičkih modela ispitivanog<br />reakcionog sistema, u oba slučaja sa maksimumom<br />relativnog prinosa epoksida kao funkcijom cilja.<br />Ispitivanje uticaja procesnih uslova, i to temperature,<br />molskog odnosa reaktanata, količine katalizatora i brzine<br />me&scaron;anja, na tok procesa epoksidovanja je bilo osnov za<br />definisanje graničnih vrednosti procesnih uslova unutar<br />kojih je tražen maksimum prinosa epoksida. Oblast dugih<br />vremena reagovanja, koja nije od interesa za industriju, je<br />izbegnuta adekvatnim izborom temperature.<br />Pri optimalnim vrednostima procesnih uslova<br />epoksidovanja lanenog ulja, određenim primenom<br />metodologije odzivne povr&scaron;ine, postignuto je dobro<br />slaganje očekivane i eksperimentalno određene vrednosti<br />maksimalnog relativnog prinosa epoksida, sa odstupanjem<br />od 3,28%.<br />Za potrebe određivanja optimalnih uslova izvođenja<br />procesa epoksidovanja biljnih ulja kori&scaron;ćenjem kinetičkih<br />modela, razvijena su tri pseudohomogena modela<br />ispitivanog trofaznog multireakcionog sistema. Pored<br />kinetike osnovnih reakcija formiranja persirćetne kiseline i&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; reakcije epoksidovanja dvostrukih veza triglicerida biljnog ulja,&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; kao i kinetike sporedne reakcije otvaranja epoksidne<br />grupe sa sirćetnom kiselinom, u predloženim modelima je<br />uzeta u obzir i raspodela sirćetne i persirćetne kiseline<br />između uljne i vodene faze sistema. Opisan je i uticaj<br />masno-kiselinskog sastava ulja, odnosno broja dvostrukih<br />veza u masno-kiselinskim lancima triglicerida, na kinetiku<br />reakcija. Za koeficijent raspodele sirćetne kiseline između<br />tečnih faza reakcionog sistema predložena je empirijska<br />korelacija koja je dala dobro slaganje izračunatih<br />vrednosti sa eksperimentalnim podacima. Kinetički<br />parametri modela su određeni fitovanjem<br />eksperimentalnih podataka o promenama količina<br />dvostruke veze i epoksidne grupe sa vremenom izvođenja<br />procesa epoksidovanja. Na osnovu statističkih pokazatelja<br />uspe&scaron;nosti fitovanja eksperimentalnih podataka, potvrđena<br />je prepostavka da je pseudohomogeni model publikovan u<br />literaturi unapređen uzimanjem u obzir pomenutih<br />fenomena raspodele komponenata reakcione sme&scaron;e i<br />masno-kiselinskog sastava sirovine pri modelovanju<br />reakcionog sistema epoksidovanja biljnih ulja<br />persirćetnom kiselinom.<br />Kori&scaron;ćenjem predloženih pseudohomogenih modela<br />reakcionog sistema za određivanje optimalnih uslova<br />izvođenja procesa epoksidovanja lanenog ulja in situ<br />formiranom persirćetnom kiselinom u prisustvu<br />jonoizmenjivačke smole, dobijeno je odstupanje od 5,51%<br />očekivane od eksperimentalno određene vrednosti<br />relativnog prinosa epoksida.<br />Bolje slaganje predviđene sa eksperimentalno određenom<br />vredno&scaron;ću relativnog prinosa epoksida u kontrolnom<br />eksperimentu je dobijeno primenom metodologije odzivne<br />povr&scaron;ine u poređenju sa kori&scaron;ćenjem kinetičkih modela pri<br />određivanju optimalnih vrednosti procesnih uslova. To je i<br />očekivano, s obzirom da regresiona jednačina kori&scaron;ćena u<br />okviru metodologije odzivne povr&scaron;ine bolje fituje relativni<br />prinos epoksida. Standardna devijacija relativnog prinos<br />epoksida za regresionu jednačinu je 8,9 puta niža od one<br />izračunate za kinetički model koji najbolje predviđa<br />optimalne procesne uslove epoksidovanja lanenog ulja<br />persirćetnom kiselinom.</p> / <p>Vegetable oils can be transformed into added value<br />products by various chemical modifications, such as<br />epoxidation. The epoxidized vegetable oils have a<br />wide range of applications in the chemical and<br />polymer industry. The quality, and consequently the<br />application, of epoxidized vegetable oil is influenced<br />by the epoxy group content. Since the epoxy groups<br />are formed by the oxidation of double bonds in<br />triglycerides, the main constituent of vegetable oils,<br />highly unsaturated vegetable oils, such as linseed<br />oil, are desirable raw materials.<br />The manufacturing of epoxidized vegetable oils<br />requires the optimization of the process conditions<br />in order to achieve complete conversion of double<br />bonds and high selectivity of the process in respect<br />to the epoxy groups. Therefore, the aim of this<br />doctoral thesis is to determine the optimal process<br />conditions for the epoxidation of linseed oil with<br />peracetic acid, formed in situ from acetic acid and<br />30% hydrogen peroxide in the presence of an ion<br />exchange resin as the catalyst. The optimal process<br />conditions were determined by response surface<br />methodology, as well as by using developed pseudohomogeneous<br />kinetic models that describe the<br />investigated reaction system. For both optimization<br />methods, the relative epoxy yield was selected as an<br />objective function to be maximized.<br />The effects of process conditions, such as<br />temperature, molar ratio of reactants, catalyst<br />amount and steering speed, on the kinetics of the<br />epoxidation were studied in order to define<br />constraints for the optimization. To avoid long<br />reaction times, which are not of interest in<br />manufacturing, an adequate temperature range was<br />selected. Under the optimized process conditions for the<br />epoxidation of linseed oil, which were determined<br />by response surface methodology, good agreement<br />between the calculated and experimentally<br />determined relative epoxy yields was achieved<br />within 3.28%.<br />Three models describing the three-phase multireaction<br />system of vegetable oil epoxidation with<br />peracetic acid were developed and further used for<br />the optimization. The models are pseudohomogeneous<br />with respect to the catalyst. Besides<br />the kinetics of the main reactions of peracetic acid<br />and epoxy group formation, the models take into<br />account the side reaction of the epoxy group opening<br />with acetic acid. The partitioning of the acetic acid<br />and peracetic acid between the oil and aqueous<br />phases is considered. In two proposed models, the<br />effect of fatty acid composition on the kinetics of the<br />process is also described by considering the number<br />of double bonds in the fatty acid chains. The<br />developed empirical correlation for the partition<br />coefficient for acetic acid between the liquid phases<br />shows good agreement between the calculated and<br />experimental data. The kinetic parameters of the<br />proposed pseudo-homogeneous models were<br />determined by fitting the experimentally determined<br />changes of the double bond and epoxy group<br />amounts with reaction time of the epoxidation.<br />Statistical values of the models` parameters<br />determination confirmed the hypothesis that the<br />pseudo-homogeneous model proposed in the<br />literature can be improved by considering the<br />partitioning phenomena and the effect of the oil fatty<br />acid composition on the kinetics of the vegetable<br />oils epoxidation with peracetic acid.<br />Under the optimized process conditions for the<br />epoxidation of linseed oil with peracetic acid formed<br />in situ in the presence of the ion exchange resin,<br />which were determined by using proposed pseudohomogeneous<br />models, the experimentally<br />determined relative epoxy yield was 5.51% lower<br />than the calculated.<br />Better agreement between the calculated and<br />experimentally determined values for the relative<br />epoxy yield, achieved under the optimal process<br />conditions, is obtained when the response surface<br />methodology (RSM) was applied as opposed to<br />when the kinetic models were used for the<br />determination of the optimal process conditions.<br />This is in accordance with better fitting of the<br />relative epoxy yield by RSM regression equation<br />than by kinetics models. Standard deviation of the<br />relative epoxy yield for RSM regression equation is 8.9 times lower than the standard deviation for the<br />most successful kinetic model used for prediction of<br />the optimal process conditions for the epoxidation of<br />the linseed oil by peracetic acid.</p>

<p>Promene komponenata i strukture mleka ispitane su tokom fermentacije kori&scaron;ćenjem inokuluma kombuhe kultivisane na crnom čaju zaslađenom saharozom<br />u koncentraciji 10%. Fermentacija mleka sa 2,2% mlečne masti izvr&scaron;ena je na dve različite temperature (37&deg;C i 42&deg;C) i uzorci su analizirani na sledećim pH vrednostima: 6,1; 5,8; 5,4; 5,1; 4,8 i 4,6. Urađena je determinacija i identifikacija produkata fermentacije laktoze kao &scaron;to su: &scaron;ećeri, organske kiseline i masne kiseline. Ispitane su teksturalne i reolo&scaron;ke karakteristike i mikrostruktura uzoraka. Pored toga, utvrđene su sličnosti i razlike uticaja inokuluma kombuhe na proces fermentacije mleka u poređenju sa delovanjem jogurtne, odnosno probiotske starter kulture.<br />Tokom fermentacije mleka u različitim fazama procesa pri odabranim pH vrednostima na 42&deg;C transformi&scaron;e se 14,6% laktoze odnosno 18,2% na 37&deg;C. Sadržaj galaktoze i glukoze koje nastaju fermentacijom i hidrolizom laktoze raste između prve i druge tačke fermentacije (pH=6,07 i pH=5,8). Dominantne organske kiseline tokom fermentacije su L&ndash;mlečna kiselina i sirćetna kiselina. Nije utvrđena značajna razlika u sadržaju masnih kiselina tokom fermentacije mleka kombuhom na 37&deg;C i 42&deg;C. Sadržaj palmitinske kiseline u mleku i uzorcima tokom fermentacije je najveći, zatim slede miristinska, stearinska i oleinska kiselina.<br />Praćenjem procesa fermentacije mleka uz primenu kombuhe na 37&deg;C i 42&deg;C od početne pH vrednosti 6,07 do zavr&scaron;ne 4,6, najznačajnije promene teksturalnih karakteristika (čvrstoće, konzistencije, kohezivnosti i indeksa viskoziteta) i reolo&scaron;kih svojstava zabeležene su između pH=5,4 i 5,1, &scaron;to je u korelaciji sa mikrostrukturom.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Promene viskoziteta u svim uzorcima pokazuju istu regresionu liniju, sa različitim koeficijentima i visokom vredno&scaron;ću r2, osim uzorka proizvedenog primenom jogurtne kulture na pH 5,4. Uzorak dobijen kori&scaron;ćenjem inokuluma kombuhe imao je najveće vrednosti kompleksnog modula, &scaron;to je rezultiralo boljim reolo&scaron;kim karakteristikama gotovog proizvoda.<br />Da bi se definisao empirijski model procesa fermentacije laktoze u mleku delovanjem kombuhe formulisana su prethodno dva matematička modela za kinetiku fermentacije saharoze u tradicionalnom kombuha napitku (na crnom čaju) &ndash; jedan koji opisuje promenu koncentracije saharoze tokom fermentacije, i drugi koji opisuje brzinu fermentacije. Promena koncentracije laktoze na ispitivanim temperaturama 37&deg;C i 42&deg;C sastoji se od dve lag faze između kojih je faza izraženog pada koncentracije. Krive zasićenja pokazuju sigmoidalnu kinetiku na nižim koncentracijama laktoze, ukazujući na kompleksni ne&ndash;Michaelis&ndash;Mentenov tip kinetike.<br />Generalno može se istaći da su primenom inokuluma kombuhe tokom fermentacije mleka u različitim fazama procesa ustanovljene specifične promene komponenata i strukture u poređenju sa efektima delovanja jogurtne odnosno probiotske starter kulture.</p><p>&nbsp;</p> / <p>Changes of milk components and structure were examined during fermentation by kombucha inoculum cultivated on black tea switened with saccharose in a concentration of 10%. The fermentation of milk with 2.2% of milk fat was carried out at two different temperatures (37&deg;C i 42&deg;C) and samples were analyzed at the following pH values: 6.1; 5.8; 5,4; 5.1; 4.8 and 4.6. Determination and identification of the lactose fermentation products, such as: sugars, organic acids and fatty acids were carried out. Textural and rheological characteristics and microstructure of the samples were investigated. Furthermore, the effect of kombucha on the milk fermentation process was compared with the effect of yoghurt and probiotic starter culture.<br />During milk fermentation at various stages of the process 14.6% of lactose content was transformed at 42&deg;C and 18.2% at 37&deg; C. The galactose and glucose content, which are formed by lactose hydrolyses and fermentation, increased between first and second pH point of fermentation (pH = 6.07 and pH = 5.8). Dominant organic acids during fermentation are L&ndash;lactic acid and acetic acid. There is no significant difference between samples in fatty acids content during fermentation on 37&deg;C and 42&deg;C. The level of palmitic acid in milk and samples was the highest of all fatty acids, followed by myristic, stearic, and oleic acid.<br />The most significant changes in textural properties (firmness, consistency, cohesiveness and viscosity index) and viscosity during milk fermentation by kombucha at 37&ordm;C and 42&deg;C, from the initial pH value 6.07 to a final 4.6, were recorded between pH=5.4 and 5.1, which is in correlation with microstructure.<br />Viscosity changes in all samples showed the same regression line with the different coefficients and a rather high r2 except for the sample produced with standard yoghurt culture at the pH 5.4. Samples produced with kombucha had the highest values of the complex modulus, which indicates better rheological characteristics of the final product.<br />In order to define an empirical model of lactose fermentation process in milk by kombucha, two mathematical models (one for the change of saccharose concentration, during its fermentation by kombucha, and the other for the rate of the mentioned fermentation) previously were formulated. Change of lactose concentrations at 37&ordm;C and 42&ordm;C consists of two retaining stages and very steep descend in-between. Saturation curves show a sigmoidal kinetics at low lactose concentrations, indicating a complex non&ndash;Michaelis&ndash;Menten type kinetics.<br />Generally it can be concluded that specific changes in the components and structure of milk by application of kombucha inoculum during fermentation at different stages of the process were established in comparison with the effects of yoghurt and probiotic starter culture.</p>

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