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V diplomskem delu je predstavljena biotehnološka proizvodnja karotenoidov in njihova možna aplikacija. Karotenoide ločimo glede na kemijsko strukturo na karotene in ksantofile. Lahko so sintetizirani v času normalne celične rasti (primarni karotenoidi) ali kot posledica neustreznih rastnih pogojev (sekundarni karotenoidi). V celici imajo zaščitno funkcijo (absorbirajo svetlobo, zadržujejo presežek energije med fotosintezo) in antioksidativno funkcijo (nevtralizacija prostih radikalov, preprečevanje oksidacije lipidov). Diplomsko delo je osredotočeno na mikroalge, ker nudijo nove možnosti in izzive ter se vedno več uporabljajo za pridobivanje pomembnih in uporabnih snovi. Obstajata dva načina kultivacije mikroalg: odprti in zaprti sistemi. V diplomskem delu je na kratko opisana tudi biosinteza karotenoidov ter načini povečanja biosinteze. Navedene in opisane so različne stopnje zaključnih procesov (ločevanje, sušenje, liza celic, ekstrakcija, čiščenje, obstojnost) in katere metode se lahko uporabljajo v ta namen. Karotenoide lahko uporabljamo v živilski industriji in v pripravi krme zaradi barvnih in prehranskih lastnosti oziroma kot aditive. Uporabljajo se tudi kot naravna barvila v kozmetiki in v farmacevtski industriji. Med novejše aplikacije sodi tudi vgrajevanje karotenoidov v polimerne produkte. The diploma thesis presents the biotechnological production of carotenoids and their possible application. Carotenoids are divided into two groups according to their chemical structure, namely carotenes and xanthophylls. They can be synthesized during normal cell growth (primary carotenoids) or due to inadequate growth conditions (secondary carotenoids). They have a protective function in the cell (they absorb light, retain excess energy during photosynthesis) and an antioxidant function (neutralization of free radicals, prevention of lipid oxidation). The thesis focuses on microalgae because they offer new opportunities and challenges and are increasingly used to obtain important and useful substances. There are two ways to cultivate microalgae, namely as an open and closed system. The diploma thesis also briefly describes the biosynthesis of carotenoids and ways to increase biosynthesis. Different stages of the downstream processes (separation, drying, cell lysis, extraction, purification, persistence) and the methods used for this purpose are listed and described. Carotenoids can be used in the food industry and in the preparation of feed due to their colour and nutritional properties or as additives. They are also used as natural dyes in the cosmetics and pharmaceutical industries. Recent applications include the incorporation of carotenoids into polymer products.

Glavni žlahtniteljski ciliji pri rastlinah so najpogosteje odpornost ali tolerantnost na bolezni in škodljivce, večja količina pridelka in boljša kakovost. Ker ima potrošnik, pri rastlinah pomembnih za človeško prehrano, raje že znane barve, sem se v tej diplomski nalogi osredotočila na novejša znanja o barvah pri okrasnih rastlinah. Zanimive barve cvetov pri okrasnih rastlinah pri potrošniku vzbujajo veliko zanimanja in predstavljajo velik pomen pri pridobivanju novih sort okrasnih rastlin. Modelni organizem za proučevanje barve cvetov je petunija. S pomočjo petunij so znanstveniki opredelili veliko število genov, ki vplivajo na barvo cvetov, vključno s tistimi, ki kodirajo biosintezo encimov, regulatorjev njihovega izražanja in funkcije vakuole. Za vidne barve listov in cvetov pri rastlinah so odgovorni različni metaboliti kot so klorofil, antociani, flavonoli in karotenoidi. Danes nam nove tehnike preurejanja genov prinašajo nove možnosti za proučevanje delovanja genov in ustvarjanje novih barv. Za najbolje prodajano rezano cvetje na svetu so že na voljo glavna genetska zaporedja in protokoli transformacije in vitro. Vsekakor pa raziskovalci še vedno veliko delajo na področju barv. V diplomskem delu se bom osredotočila na predstavitev ključnih barv, dejavnikov, ki vplivajo na barve, njihovih genov pri nekaterih vrstah okrasnih rastlin in možnosti njihove manipulacije. The main breeding objectives for plants are most often resistance or tolerance to diseases and pests, increased yield and improved quality. As consumers prefer familiar colours in plants important for human consumption, I have focused in this thesis on more recent knowledge on colours in ornamental plants. Interesting flower colours in ornamental plants are of great interest to the consumer and are of great importance in the acquisition of new ornamental plant species. The model organism for the study of flower colour is the petunia. Using petunia, scientists have identified a large number of genes that influence flower colour, including those encoding biosynthetic enzymes, regulators of their expression and vacuole function. Various metabolites such as chlorophyll, anthocyanins, flavonols and carotenoids are responsible for the visible colours of leaves and flowers in plants. Today, new gene-editing techniques provide us with new opportunities to study how genes work and to create new colours. For the world's best-selling cut flowers, the main genetic sequences and in vitro transformation protocols are already available. However, there is still much work to be done by researchers in the field of colour. In my thesis I will focus on the presentation of key colours, the factors influencing colours, their genes in some varieties of ornamental plants and the possibilities of their manipulation.

Vitamin A je skupina v maščobi topnih spojin, ki jih človek sam ni zmožen sintetizirati, zato morajo biti široko prisotne v človeški prehrani. Če pride do pomanjkanja vitamina, lahko to vodi v številne težave, med drugim tudi do slepote, kar poleg lakote predstavlja precej velik problem v večini držav v razvoju, kjer škrobna prehrana prebivalstva ne vsebuje zadostnih količin omenjenega vitamina. V ta namen so znanstveniki razvili številne kulturne rastline, ki imajo povečano sintezo in kopičenje provitamina A, ki se v človeškem organizmu pretvori v funkcionalen vitamin A. To so dosegli z metodami biofortifikacije (s konvencionalnim križanjem in z genetskim inženiringom kulturnih rastlin) in s pomočjo 2 pristopov: prekomerne ekspresije in utišanja genov. Uporabljene metode so privedle do znatnega kopičenja karotenoidov v užitnih delih riža, paradižnika, krompirja in koruze ter do očitne spremembe barve plodov le-teh, ki so z večjo vsebnostjo karotenoidov kazale bolj intenzivne odtenke oranžne oz. rumene barve. Za uspešno manipulacijo karotenogeneze pa je ključnega pomena temeljito razumevanje biosintezne poti karotenoidov, katere poznavanje je igralo ključno vlogo pri izbiri posameznih genov. Pri opisanih kulturnih rastlinah so bili geni predvsem bakterijskega izvora (rod Erwinia), uporabljeni pa so bili tudi rastlinski geni, tako endogeni kot tudi geni iz drugih rastlinskih vrst, ki že same po sebi vsebujejo večje količine karotenoidov. Vitamin A is a group of fat-soluble compounds that humans cannot synthesize themselves, so they must be widely present in the human diet. If there is a deficiency of the vitamin, it can lead to many problems, including blindness, which, in addition to hunger, is a big problem in most developing countries where the starchy diet of the population does not contain sufficient amounts of said vitamin. For this purpose, scientists have developed several cultivated crops that have increased synthesis and accumulation of provitamin A, which is converted into functional vitamin A in the human body. This was achieved by biofortification methods (conventional crossing and genetic engineering of cultivated crops) and by means of 2 approaches: overexpression and gene silencing. The methods used led to a significant accumulation of carotenoids in the edible parts of rice, tomatoes, potatoes and corn, and to an obvious change in the colour of their fruits, which, with a higher content of carotenoids, showed more intense shades of orange or yellow colour. For the successful manipulation of carotenogenesis, a thorough understanding of the biosynthesis pathway of carotenoids is crucial and played a key role in the selection of individual genes. In the cultivated plants described, the genes were mainly of bacterial origin (genus Erwinia), but plant genes were also used, both endogenous and genes from other plant species, which by themselves contain larger amounts of carotenoids.

Kopriva (Urtica dioica L) kao samonikla, ali i kultivirana biljna vrsta, zbog sastava i udjela različitih bioaktivnih spojeva ima značajna biološka svojstva te potencijalna pozitivna djelovanja u prevenciji i ublažavanju simptoma različitih oboljenja. Ljekovita svojstva koprive pripisuju se sekundarnim metabolitima poput karotenoida, klorofila, vitamina, polifenola koji su najvećim dijelom zastupljeni u nadzemnom dijelu biljke, dok su spojevi poput fitosterola zastupljeni u podzemnom dijelu odnosno korijenu. Obogaćena je riboflavinom, askorbinskom kiselinom, folnom kiselinom, pantotetskom kiselinom te aminokiselinama, a u samom korijenu koprive pronađene su veće količine cisteina, glicina i triptofana. U okviru ovog rada istaknute su i opisane najznačajnije skupine bioaktivnih spojeva zastupljenih u nadzemnom dijelu koprive (polifenoli, karotenoidi, klorofili) te u korijenu (fitosteroli), kao i njihova povezanost s ljekovitim svojstvima ove biljke i njezinih proizvoda.

Research background. The industrial transformation of tropical fruits, and in particular guava, generates a large quantity of by-products that are generally disposed of as organic waste. In these by-products a large quantity of bioactive substances is concentrated, such as carotenoids, which can be used for the partial substitution of nitrites in meat sausages without affecting their physicochemical, colour and conservation characteristics. Although there are some studies in this regard, there is still a lack of research, especially on guava residues, to verify this hypothesis. Therefore, the aim of this study is to investigate the influence of the components of the guava epicarp extract on the physicochemical, textural, colour and sensory properties of frankfurters. Experimental approach. Three treatments were investigated (25, 50 and 75% nitrite replacement with guava epicarp flour extract), along with a control treatment without the extract. The physicochemical properties, colour coordinates, and texture parameters were analysed, and a sensory evaluation was also carried out. The treatment that gave the best results was subjected to stability analysis over 30 days. Results and conclusions. The addition of 25% guava epicarp flour extract gave the best results, since it did not affect the colouration of the sausages or their physicochemical and textural properties. Likewise, during the stability analysis over time, the parameters related to fat oxidation were not affected, and final products had a residual nitrite load of (23.6±0.2) mg/kg, lower than the maximum allowed (150 mg/kg). Novelty and scientific contribution. The partial substitution of the nitrites in frankfurters with the carotenoids from the guava epicarp flour added in a mass fraction up to 25% can be a viable alternative to produce sausages with lower content of nitrites, without affecting their colouration or shelf life. This is important for the meat industry, which favours the development of new products using substances of natural origin., Pozadina istraživanja. Industrijska obrada tropskog voća, osobito guave, proizvodi veliku količinu nusproizvoda, koji se uglavnom otklanjaju kao organski otpad. Ti nusproizvodi sadržavaju velike količine bioaktivnih tvari, kao što su karotenoidi, koji bi se mogli upotrijebiti kao djelomična zamjena za nitrite u kobasicama, a da pritom ne narušavaju njihova fizikalno-kemijska svojstva, boju i trajnost. Iako postoje neka istraživanja na tu temu, obrada otpada guave nije dovoljno istražena da bi potvrdila ovu pretpostavku. Stoga je svrha ovoga rada bila ispitati utjecaj sastojaka ekstrakta epikarpa guave na fizikalno-kemijska, teksturalna i senzorska svojstva te boju hrenovki tipa Frankfurter. Eksperimentalni pristup. Ispitana su tri uzorka (u kojima je 25, 50 ili 75 % nitrita zamijenjeno ekstraktom brašna dobivenim iz epikarpa guave), te kontrolni uzorak bez dodatka ekstrakta. Ispitana su fizikalno-kemijska svojstva, koordinate boje i parametri teksture, te je provedena senzorska analiza kobasica. Njihova stabilnost tijekom 30 dana ispitana je na uzorku koji je imao najbolje značajke. Rezultati i zaključci. Najbolji su rezultati dobiveni zamjenom 25 % nitrita ekstraktom brašna epikarpa guave, pri čemu nije došlo do promjene boje niti fizikalno-kemijskih i teksturalnih svojstava hrenovki. Također nije došlo do promjene parametara oksidacije masti tijekom skladištenja, a krajnji je proizvod sadržavao (23,6±0,2) mg/kg nitrita, što je niže od maksimalno dozvoljene vrijednosti (150 mg/kg). Novina i znanstveni doprinos. Djelomična zamjena nitrita u hrenovkama karotenoidima iz brašna od epikarpa guave dodanog umjesto 25 % masene frakcije nitrita može poslužiti kao alternativa u proizvodnji hrenovki sa smanjenim udjelom nitrita, bez da se pritom utječe na boju i trajnost kobasica. Ovaj rezultat je važan za mesnu industriju, koja teži razvoju novih proizvoda s dodatkom prirodnih sastojaka.

Cilj ovog istraživanja bio je odrediti sadržaj karotenoida i tokoferola u ulju ploda dvije sorte pasjeg trna (PT) te ispitati utjecaj ekstrakcije potpomognute ultrazvukom (EPUzv) na izolaciju fenolnih spojeva (FS) i vitamina C iz pogače PT primjenom različitih vodenih otopina etanola (30 i 70 %, v/v) i vremena ekstrakcije (20, 30, 40 min). Veći antioksidacijski kapacitet (AOK) i viši maseni udjeli α- tokoferola i karotenoida određeni su u ulju sorte Leikora, a najzastupljeniji karotenoida bili su lutein i zeaksantin te njihovi derivati. Najveće vrijednosti vitamina C i ukupnih FS određene su u 70 %-tnim etanolnim ekstraktima pogače sorte Ascola (66,02 mg/100 g) nakon 20 odnosno 40 min ekstrakcije te AOK u pogači sorte Leikora nakon 40 min ekstrakcije. Najzastupljeniji FS u pogači PT su hidroksicimetne kiseline (36,32 mg/100 g s. tv.) te je bolja učinkovitost EPUzv postignuta primjenom 70 %-tne vodene otopine etanola i dužeg vremena ekstrakcije. The aim of the study was to determine the carotenoid and tocopherol content in two fruit oil varieties of sea buckthorn (SBO) and to evaluate the effect of ultrasound-assisted extraction (UAE) on the isolation of phenolic compounds (PC) and vitamin C from pomace (SBP) using different ethanol solutions (30 and 70%, v/v) and extraction times (20, 30, 40 min). Higher mass fractions of α-tocopherol and carotenoids and higher antioxidant capacity (AOC) were found in Leikora SBO. The most abundant carotenoids were lutein and zeaxanthin and their derivatives. The highest vitamin C and total PC content were determined in 70% ethanol extracts of Ascola SBP and extraction times of 40 and 20 min, while the highest AOC was determined in Leikora SBP after 40 min. Hydroxycinnamates (36.32 mg/100 g dm) were the most abundant PC and better UAE efficiency was obtained using 70% ethanol solution and longer extraction time.

Bakterije iz rodu Streptomyces so proizvajalci medicinsko in industrijsko pomembnih sekundarnih metabolitov. Genom streptomicet zapisuje biosintezne genske gruče (BGG) za več kot 20 različnih sekundarnih metabolitov, vendar večino teh metabolitov, ni mogoče odkriti pod standardnimi laboratorijskimi pogoji. Izražanje t.i. tihih BGG v heterolognih gostiteljih je privlačen pristop za reševanje tega problema. Številni streptomcetni gostitelji že obstajajo, vendar ne dosegajo zadostnih koncentracij ciljnega produkta. Streptomyces rimosus je pomemben industrijski proizvajalec, ki je znan po doseganju visokih koncentracij antibiotika oksitetraciklina (OTC). Naš cilj je bil oceniti sposobnost S. rimosus kot gostitelja za proizvodnjo heterologno izraženih produktov. Najprej smo uspešno odstranili dve BGG, ki kodirata OTC in izorenieraten, s čimer smo zmanjšali tekmovanje med nativnimi in heterolognimi BGG. Razvili smo integrativne vektorje, ki vsebujejo BGG, ki kodirajo biosintezo tetraciklina kelokardina (CHD), izoprenoidov ciklooktatina in β-karotena ter večkrat nenasičenih maščobnih kislin (PUFA). Vsi našteti metaboliti uporabljajo malonil-CoA kot glavni gradnik. Poleg nativne CHD BGG smo rekonstituirali tudi sintetično BGG. Poleg izoprenoidne BGG smo klonirali tudi mevalonatno pot in jo vnesli v S. rimosus, s čimer smo zagotovili boljšo razpoložljivost izoprenoidnega substrata. Za povečanje donosa PUFA smo v S. rimosus uvedli gen, ki kodira fosfopanteteinil transferazo. Ciklooktatin, β-karoten in PUFA BGG so bili uspešno izraženi v S. rimosus, medtem ko niti nativna niti sintetična BGG za kelokardin nista proizvajala ciljnega produkta. Streptomyces are producers of medically and industrially important secondary metabolites. The genome of Streptomyces encodes biosynthesis of more than 20 different secondary metabolites. However, most of the secondary metabolites encoded by the corresponding biosynthetic gene clusters (BGCs) cannot be detected under standard laboratory conditions. Therefore, expression of these “silent” BGCs in a heterologous host seems an attractive approach to solve this problem. Several Streptomyces hosts already exists, but they do not achieve sufficient concentrations of the target product. Streptomyces rimosus is a producer of important antibiotic oxytetracycline (OTC) and is known to achieve high titers of it. Our goal was to evaluate ability of S. rimosus as a host to produce heterologous products. First, we successfully removed two BGCs encoding OTC and isorenieratene, reducing competition between native and heterologously expressed BGCs. We developed integrative vectors containing BGCs encoding tetracycline chelocardin (CHD), isoprenoids cyclooctatin and β-carotene, and polyunsaturated fatty acids (PUFA), all of which consume malonyl-CoA as a main building block. In addition to native CHD BGC, we also reconstituted synthetic BGC. In addition to the isoprenoid BGC, we also cloned the mevalonate pathway and introduced it into S. rimosus, which could ensure better availability of isoprenoid substrate. To increase yield of PUFA, the gene encoding phosphopantetheinyl-transferase was introduced into S. rimosus. Cyclooctatin, β-carotene and PUFA BGCs were successfully expressed in S. rimosus, whereas neither native nor synthetic chelocardin BGC resulted in the production of the target product.

V okviru doktorskega dela smo identificirali in kvantificirali karotenoide v ekstraktih listov v različnih rastnih fazah dveh invazivnih tujerodnih rastlinskih vrst – japonski dresnik (Fallopia japonica Houtt) in češki dresnik (Fallopia x bohemica). Karotenoidni profili so bili za obe rastlinski vrsti zelo podobni. Ugotovili smo, da se količina prostih ksantofilov in karotenov v jesensko obarvanih listih bistveno zmanjša, po drugi strani pa smo pokazali, da se pojavijo nove ksantofilne spojine – ksantofilni estri maščobnih kislin. Zeleni listi obeh preučevanih dresnikov predstavljajo bogat in trajnosten naravni vir bioaktivnih karotenoidov, predvsem β-karotena (69–97 mg/100g s.m.) in prostega luteina (97–144 mg/100g s.m.), ki bi jih lahko uporabili za pripravo prehranskih dopolnil. Obstaja pa težava. Ker so karotenoidi izven rastlinske matrice zelo nestabilni in v prisotnosti povišane temperature, svetlobe, oksidantov, kislin ali kovin hitro razpadejo, je njihova uporaba omejena. Navdih za reševanje tega problema smo dobili s posnemanjem naravnih procesov, namreč, estrenje fotosintetskih ksantofilov naj bi v jeseni med drugim povečalo njihovo kemijsko stabilnost. V ta namen smo razvili prvo okolju prijazno in ekonomsko upravičljivo sintetsko platformo, kjer smo uporabili zelene, GRAS in obnovljive reagente ter topila. V β-pinenu kot reakcijskem topilu in pri sobni temperaturi ter atmosferskem tlaku smo s kombinacijo 5 modelnih ksantofilov in 11 strukturno in elektronsko različnih kislinskih anhidridov pripravili 55 različnih ksantofilnih estrov. Pokazali smo, da se estrenje lahko izvede tudi brez topila in z uporabo ekstrakta odpadnega rastlinskega materiala zelenih listov japonskega dresnika in olupkov avokada, ki predstavljata bogat obnovljiv vir luteina oziroma anteraksantina. Sintetizirane spojine smo očistili in jih okarakterizirali na podlagi podatkov pridobljenih s pomočjo HPLC-PDA-MSn in HRMS. Od 55 spojin jih je bilo 22 prvič uspešno sintetiziranih. Uspešnost povečanja kemijske stabilnosti ksantofilov preko estrenja smo preverili na 13 različnih luteinskih diestrih, ki smo jih sintetizirali bodisi iz čistega luteina bodisi iz ekstrakta zelenih listov japonskega dresnika. Vsak luteinski diester smo ločeno izpostavili posameznemu stresnemu pogoju: povišani temperaturi (60 °C), svetlobi (UV-A), oksidantu (H2O2) in kislemu okolju. Na podlagi sistematične stabilnostne študije smo prvi pokazali povezavo med kemijsko stabilnostjo, kemijsko strukturo in okoljem luteinskih estrov, v katerem se nahajajo. Od vseh pripravljenih estrov so se za najbolj obetavne izkazali lutein di(2,2-dimetilpropanoat), lutein di(2-metilpropanoat) in lutein di(3- metilbutanoat), saj so bili, ne glede na izbrani stresni pogoj, 1,4–20-krat bolj stabilni v primerjavi s prostim luteinom. Ugotovili smo, da lahko matrica rastlinskega ekstrakta močno vpliva na stabilnost ksantofilov (pozitivno ali negativno), odvisno od stresnega pogoja in tipa estra. Listi japonskega dresnika torej predstavljajo odpadek, ki na trajnosten način omogoča pripravo potencialnih izdelkov z visoko dodano vrednostjo – ksantofilnih estrov s povečano kemijsko stabilnostjo. Carotenoids in mature (green) and senescing leaves of invasive alien plant species Japanese (Fallopia japonica Houtt) and Bohemian (Fallopia x bohemica) knotweed were identified and quantified. Both plants showed similar pigment profiles.The amount of free xanthophylls and carotenes was significantly lower in senescing leaves in comparison to green leaves. On the other hand new xanthophyll compounds appeared in senescing leaves – xanthophyll fatty acid esters. Particularly green leaves of both studied knotweeds represent a rich and sustainable natural source of bioactive carotenoids, mainly β-carotene (69–97 mg/100g D.W.) and xanthophyll lutein (97–144 mg/100g D.W.), thus exploitation of these invaders can be used for the production of high added-value products. However, due to carotenoids chemical structure, they are very unstable outside of the plant matrix and they degrade fast in the presence of elevated temperature, light, oxidants, acids and metals. We found an inspiration to solve this problem in nature itself, where in autumn xanthophylls are converted into more chemically stable xanthophyll esters. To this end, we developed the first environmentally friendly and economically viable synthetic platform, where overall green, renewable, GRAS and recyclable materials were used. The synthesis was carried out in β-pinene as the reaction solvent and at ambient conditions (room temperaure, Atmospheric pressure), where 55 different xanthophyll esters (combining 5 model xanthophylls with 11 structurally and electronically distinct acid anhydrides) were prepared. We have shown that esterification can be performed also without use of a solvent and by exploitation of a waste plant material, e.g. extract of green leaves of Japanese knotweed or avocado peels, which represent rich renewable source of lutein and anteraxanthin, respectively. The synthesized compounds were purified and characterized by means of HPLC-PDA-MS and offline HRMS. Out of 55 compounds, 22 were successfully synthesized for the first time. The efficiency of our approach to achieve higher chemical stability of xanthophylls via esterification was tested on 13 different lutein diesters, which were synthesized either from pure lutein or from green leaves extract of Japanese knotweed. Each individual lutein diester was separately examined at each individual stress conditions: elevated temperature (60 °C), light (UV-A), oxidant (H2O2) and acidic environment. Based on systematic stability studies, we were first to identify a relationship between the chemical stability, chemical structure and the environment by which lutein esters were surrounded. Of all the prepared lutein esters, lutein di(2,2-dimethylpropanoate), lutein di(2-methylpropanoate) and lutein di(3- methylbutanoate) proved to be the most stable. They showed to be 1,4–20 times more stable than free lutein, regardless of the chosen stress condition. Depending on the stress condition the plant matrix can strongly (positively or negatively) affect the stability of an individual xanthophyll. The green leaves of Japanese knotweed, therefore, represent a suitable plant waste material that enables the preparation of potential high value-added products – xanthophyll esters with increased chemical stability.

Beta karoten, likopen i kurkumin su biljni pigmenti koji pokazuju veliki antioksidacijski potencijal. Bitni su i često korišteni u održavanju ljudskog zdravlja i sprječavanju nastanka bolesti. Zbog svog antioksidacijskog djelovanja smatraju se nutraceuticima – vrijednim sastavnicama novih oblika enteralne prehrane. Enteralna prehrana, zbog svoje uloge dopunjavanja ljudske prehrane u stanjima i bolestima kada je to kompromitirano kroz zdravu prehranu, sadržajno je bogata makronutrijentima i mikronutrijentima. Prehrambena vlakna su bitna sastavnica koja osigurava zdravu probavu i može djelovati probiotički, no zbog svojih fizikalno-kemijskih svojstava stupaju u interakcije s karotenoidima i kurkuminom te mogu smanjiti njihovu biodostupnost. U ovom diplomskom radu ispitana je stabilnost beta karotena, likopena i kurkumina u simuliranim uvjetima probave te biodostupnost s 10 mješavinama prehrambenih vlakana. Beta karoten i likopen pokazali su stabilnost pri simulaciji uvjeta probave, dok se koncentracija kurkumina značajno smanjila (na oko 70%). Likopenu je svih 10 ispitivanih mješavina prehrambenih vlakana smanjilo biodostupnost, dok je biodostupnost beta karotena i kurkumina s nekim mješavinama ostala nepromijenjena. Dvije standardne mješavine STAND-2 i STAND-3 imaju značajan utjecaj na smanjenje biodostupnosti sva tri ispitivana spoja. Inovativne mješavine imaju različiti utjecaj na spojeve, ovisno o njihovom sastavu. Sve inovativne mješavine imaju mali utjecaj na biodostupnost beta karotena. Najveća biodostupnost likopena je s mješavinom INOV-1B (98,3%). Najveća biodostupnost kurkumina je uz mješavinu STAND-1 (113,7%). Dobiveni rezultati upućuju na sličnost dviju standardnih mješavina (STAND-2 i STAND-3), a različite sastave ostalih 8 mješavina te različite afinitet prema vezanju ispitivanih spojeva ovisno o sastavu mješavina. Beta carotene, lycopene and curcumin are plant pigments that show great antioxidant potential. They are important in maintaining human health and preventing diseases. Due to their antioxidant effect, they are considered nutraceuticals - valuable components of new formulas of enteral nutrition. Enteral nutrition, due to its role of supplementing human nutrition in conditions and diseases that compromise ingestion through a healthy diet, is rich in macronutrients and micronutrients. Dietary fibers are an essential component that ensures healthy digestion and can have a probiotic effect, but due to its physical and chemical properties, its interaction with carotenoids and curcumin can reduce their bioavailability. The stability of beta carotene, lycopene, and curcumin in simulated digestive conditions and their bioavailability as affected by different dietary fibers mixtures were examined in this master thesis. Beta carotene and lycopene showed stability during the simulation of digestion conditions, while the concentration of curcumin decreased significantly (to about 70%). The bioavailability of lycopene was reduced by all 10 dietary fiber mixtures that were analyzed, while the bioavailability of beta-carotene and curcumin remained unchanged when analyzed with some of 10 mixtures. Two standard mixtures STAND-2 and STAND-3 have a significant effect on reducing the bioavailability of all three analyzed compounds. Innovative mixtures have different effects on compounds, depending on their composition. All innovative mixtures have little effect on the bioavailability of beta-carotene. The highest bioavailability of lycopene was shown with the mixture INOV-1B (98.3%). The highest bioavailability of curcumin was shown with the STAND-3 mixture (113.7%). With obtained results it can be concluded there is similarity between two standard mixtures (STAND-2 and STAND-3), and there is difference between composition of the other 8 mixtures with different affinity towards the binding of analyzed compounds.

Koncept korištenja otpalog jesenjeg lišća kao hranidbenog suplementa za pigmentaciju u peradi oskudno ili uopće nije istražen. Sa ciljem detaljnijeg istraživanja spomenutog koncepta provedeno je istraživanje u kojem je određena koncentracija karotenoida pet biljnih vrsta: srebrnolisni javor (Acer saccharinum), likvidambar (Liquidambar styraciflua), perzijska parocija (Parrotia persica), hrast lužnjak (Quercus robur) i sitnolisna lipa (Tilia cordata). Utvrđeno je da se otpalo lišće sakupljenih biljaka razlikuje u karotenoidnom profilu pri čemu se po visokom sadržaju ukupnih karotenoida ističu srebrnolisni javor i sitnolisna lipa. Ukoliko se otpalo jesenje lišće istraživanih vrsta koristi kao izvor pigmenata u potpunoj krmnoj smjesi, nakon sušenja i mljevenja, pridonosi unosu karotenoida kod kokoši nesilica. Međutim, potrebna su daljnja istraživanja kako bi se potvrdila prikladnost otpalog jesenjeg lišća u hranidbi peradi sa strane mikrobiološkog i higijenskog aspekta.

Lamb's lettuce is a popular winter salad, often grown in private vegetable plots, small local farms or in intensive vegetable production. It is usually marketed as a ready-to-eat produce in supermarkets. The aim of the study was to evaluate the changes in biochemical composition and degradation of bioactive compounds during consumer-relevant time of home-grown and store-bought Valerianella locusta “Vit” salad. Primary metabolites, assimilatory pigments as well as secondary metabolites were monitored during 1 week of refrigerated storage. Home-grown lamb's lettuce exhibited highest levels of total sugars, total organic acids, vitamin C, and total phenolic content as well as enhanced levels of most individual phenolic compounds and chloroplast pigments. Locally produced samples of lamb's lettuce also contained high levels of analyzed bioactive components. All samples retained most bioactive components during the entire period of refrigerated storage. The results underline the instability of vitamin C during refrigerated storage of lamb's lettuce and pinpoint this parameter as being the most affected by storage.

V raziskavi smo določali profil karotenoidov v cianobakteriji Arthrospira platensis in mikroalgi Chlorella vulgaris ter vpliv mlečnokislinske fermentacije na prisotnost karotenoidov in fenolnih spojin. Pripravili smo ekstrakte past biomas A. platensis in C. vulgaris v dveh različnih topilih (100-% aceton, kloroform : metanol = 1 : 2). V ekstraktih smo karotenoide ovrednotili s spektrofotometrično metodo in s tankoplastno kromatografijo. V raziskavi smo ugotovili, da se vsebnost karotenoidov in ustreznost topil, ki sta se uporabljali pri A. platensis in C. vulgaris, razlikujeta, saj je aceton dal boljše rezultate za vzorce A. platensis in mešanica kloroform : metanol za vzorce C. vulgaris. Iz kontrolne biomase A. platensis smo s 100-% acetonom ekstrahirali 4,37 ± 0,16 (mg/g suhe biomase) karotenoidov in iz fermentirane 4,37 ± 0,07 (mg/g suhe biomase) karotenoidov. Z mešanico kloroform : metanol smo iz kontrolne biomase A. platensis ekstrahirali 25,9 % manj karotenoidov in iz fermentirane biomase 13,3 % manj karotenoidov. Iz kontrolne in fermentirane biomase C. vulgaris smo s 100-% acetonom ekstrahirali skoraj 10-krat manj skupnih karotenoidov, in sicer 0,45 ± 0,01 (mg/g suhe biomase) in 0,52 ± 0,02 (mg/g suhe biomase). Tudi z mešanico kloroform : metanol smo ekstrahirali manj karotenoidov, tako iz kontrolne biomase C. vulgaris kot fermentirane, in sicer 2,27 ± 0,12 (mg/g suhe biomase) in 1,39 ± 0,02 (mg/g suhe biomase). S tankoplastno kromatografijo smo ločili mešanico pigmentov v ekstraktih iz past kontrolnih in fermentiranih biomas. Pri A. platensis smo uspeli ločiti 10-16 komponent več z uporabo mešanice 1 kot z uporabo drugih mešanic, medtem ko smo pri C. vulgaris uspeli ločiti 4-13 komponent več z uporabo mešanice 3 kot z uporabo preostalih mešanic topil. S Folin-Ciocalteujevo metodo smo določali tudi vsebnost fenolnih spojin. Fenolne spojine predstavljajo od najmanj 0,02 % do največ 0,44 % biomase. Ugotovili smo, da je koncentracija skupnih fenolnih spojin višja v fermentiranih vzorcih kot v kontrolnih ter da A. platensis na splošno vsebuje več karotenoidov in skupnih fenolnih spojin (SFS) kot C. vulgaris. In this research, we determined profiles of carotenoids in cyanobacteria A. platensis and microalgae C. vulgaris, the effect of lactic acid fermentation on present carotenoids, and also the presence of phenolic compounds. Biomass paste extracts of A. platensis and C. vulgaris were prepared in two different solvents (100 % acetone, chloroform : methanol = 1 : 2) so that we can compare the results and see which solvent is giving better results. After extraction, we tried to determine carotenoids by various methods, such as spectrophotometry and thin-layer chromatography. In addition to carotenoids, the presence of phenolic compounds was also determined using the Folin-Ciocalteu method. In this study, we found that the carotenoid content and solubility of the solvents used differed between A. platensis and C. vulgaris, as one solvent gave better results for A. platensis samples and the other solvent for C. vulgaris samples. 4.37 ± 0.16 (mg/g d. m.) was extracted from unfermented biomass of A. platensis in 100 % acetone and 4.37 ± 0.07 (mg/g d. m.) from fermented biomass. 3.24 ± 0.53 (mg/g d. m.) was extracted from unfermented biomass of A. platensis in chloroform : methanol solvent mixture and 3.79 ± 0.25 (mg/g d. m.) of fermented biomass. When it comes to A. platensis we extracted 14–22 % more carotenoids with 100 % acetone than with chloroform : methanol solvent mixture. 0.45 ± 0.01 (mg/g d. m.) was extracted from unfermented biomass of C. vulgaris in 100 % acetone and 0.52 ± 0.02 (mg/g d. m.) from unfermented biomass. 2.27 ± 0.12 (mg/g d. m.) was extracted from unfermented biomass of C. vulgaris in chloroform : methanol solvent mixture and 1.39 ± 0.02 (mg/g d. m.) of fermented biomass. When it comes to C. vulgaris we extracted 2–4 % more carotenoids with chloroform : methanol solvent mixture than with 100 % acetone. The same was observed in thin layer chromatography by comparing the suitability of four solvent mixtures. In A. platensis a non-polar solvent mixture 1 separated 10-16 more components compared to any of the other mixtures used and in C. vulgaris a polar solvent mixture 3 separated 4-13 more components compared to other solvent mixtures. We also used Folin-Ciocalteu method to determine the content of phenolic compounds. Phenolic compounds represent from at least 0.2 % to the maximum of 0.44 % of biomass. Finally, we concluded that the concentration of total phenolic compounds and carotenoids is generally higher in fermented samples than in unfermented samples and that A. platensis generally contains more carotenoids and total phenolic compounds (TPC) than C. vulgaris.

V raziskavi smo določili vsebnost bioaktivnih snovi v sedmih različnih vrstah samoniklih travniških rastlin: navadni rman (Achillea millefolium L.), navadna česnovka (Alliaria petiolata Cav. & Grande), mehki osat (Cirsium oleraceum (L.) Scop.), navadni jajčar (Leontodon hispidus L.), veliki trpotec (Plantago major L.), navadna črnoglavka (Prunella vulgaris L.) in dišeča vijolica (Viola odorata L.). Rastlinski material smo nabrali v spomladanskem času, v enakih vremenskih razmerah, v razvojni fazi, v kateri te rastline nabirajo za prehrano. Izmerili smo vsebnost klorofilov a in b, tokoferolov in in karotenoidov: -karoten, -karoten, lutein, neoksantin, violaksantin, anteraksantin in zeaksantin. Vzorce smo analizirali s pomočjo visokoločljivostne tekočinske kromatografije (HPLC). Največjo povprečno vrednost skupnega klorofila in največjo povprečno vrednost karotenoidov smo izmerili v navadni česnovki, največjo povprečno vrednost skupnih tokoferolov pa v velikem trpotcu. Za boljšo predstavo o vsebnosti bioaktivnih snovi v preučevanih rastlinah smo primerjali naše rezultate z literaturnimi podatki za te rastline in za zelenjavo, ki jo uživamo na enak način (mladi listi). The aim of this study was to determine lipophilic antioxidants content in seven grassland plant species: Yarrow (Achillea millefolium L.), garlic mustard (Alliaria petiolata Cav. & Grande), cabbage thistle (Cirsium oleraceum (L.) Scop.), rough hawkbit (Leontodon hispidus L.), broadleaf plantain (Plantago major L.), common selfheal (Prunella vulgaris L.) and sweet violet (Viola odorata L.). Plant material was all harvested in spring, at same weather conditions, at developmental stage that is prefered from forager and consumer's standpoint. Chlorophyll a and b, tocopherols ( and  and carotenoids: lutein, neoxanthin, violaxanthin, antheraxanthin and zeaxanthin contents, were determined quantitatively. Analysis was conducted with a high performance liquid chromatography (HPLC) method. The highest average value of total chlorophyll and carotenoid contents was measured in garlic mustard, the highest average value of total tocopherol was measured in broadleaf plantain. To get a better perception of the bioactive compounds amount in studied plants, we compared results from our study to the ones from the literature for the same plant species and for other species that are being consumed in the same way as those from our study (leafy greens).

Karotenoidi, vrijedne sastavnice ljudske prehrane, poznati i kao prirodni pigmenti pokazuju visok potencijal u održavanju ljudskog zdravlja. Provedena su brojna istraživanja koja nerijetko daju drugačije rezultate i zaključke, ali potencijal beta-karotena kao zaštitnika u očuvanju zdravlja sve više raste pa se tako ističu njegova antioksidativna svojstva ali i njegov utjecaj na kožu, zdravlje očiju i kognitivne funkcije. Karotenoidi su vrijedan izvor pigmenata i vitamina, te je razvoj učinkovitih metoda njihove proizvodnje istraživačima od velikog interesa. Najčešće korištene metode za proizvodnju jednog od glavnih karotenoida, beta-karotena, su fizikalno-kemijske, kemijske i mikrobiološke metode. Kao sirovine koriste se i biljke i mikrobi. Od biljaka se najčešće koriste mrkva, batat, špinat, bundeva i rajčica, a od mikroba alga Dunaliella spp. i bakterije Rhodococcus maris i Rhodobacter sphaeroides. Konceptualni modeli omogućavaju lakše shvaćanje raznih procesa pa se tako mogu primijeniti i kod razumijevanja metabolizma karotenoida. Carotenoids, valuable components of the human diet, also known as natural pigments show high potential in maintaining human health. Numerous studies have been conducted that often give different results and conclusions, but the potential of beta-carotene as a protector in maintaining health is growing, thus highlighting its antioxidant properties but also its impact on skin, eye health and cognitive functions. Since carotenoids are a valuable source of pigments and vitamins development of effective methods of their production is of great interest to researchers. Most commonly used methods for the production of one of the major carotenoids, beta-carotene, are physicochemical, chemical, and microbiological methods. Plants and microbes are also used as raw materials. From plants, most commonly used are carrots, sweet potatoes, spinach, pumpkin and tomatoes, while from microbes’ algae Dunaliella spp. and the bacteria Rhodococcus maris and Rhodobacter sphaeroides. Conceptual models make it easier to understand various processes, so they can be applied to understand the metabolism of carotenoids.

Zbog bogatog nutritivnog sastava, niske kalorijske vrijednosti i povoljnog učinka na zdravlje čovjeka bundeva se smatra funkcionalnom hranom. Obiluje polifenolnim spojevima i karotenoidima kojima je dokazano antioksidacijsko djelovanje. Za konzerviranje i produljenje trajnosti bundeve često se koristi sušenje, najstarija metoda konzerviranja hrane. Sušenje može značajno utjecati na nutritivni sastav, a posebno na antioksidacijsku aktivnost namirnice. Cilj ovog završnog rada je prema rezultatima do sada objavljenih znanstvenih radova prikazati utjecaj pojedinih procesa sušenja na antioksidacijsku aktivnost bundeve. Najčešće korišteni procesi sušenja bundeve su konvekcijsko, mikrovalno, osmotsko i sušenje zamrzavanjem, a često se koristi i kombinacija navedenih procesa. Rezultati ukazuju da je uz izbor odgovarajućeg procesa, kao i uvjeta sušenja, moguće zadržati antioksidacijska svojstva bundeve nakon sušenja. Valuable nutritive composition, low calories and great benefits for human health classifies pumpkin as functional food. It is rich in polyphenols and carotenoids, compounds with proved antioxidant activity. Drying, the oldest method for food preservation is very often used to prolong shelf life and pumpkin preservation. Drying may affect nutritive composition, especially antioxidant activity of food. According previous published scientific data, the aim of this bachelor thesis is to present results about impact of some used processes for pumpkin drying on antioxidant activity of pumpkin. Most commonly used processes for pumpkin drying are convective, microwave, osmotic and freeze drying. Very often is used the combination of these processes. Results indicate that appropriate drying process, as well as drying conditions can preserve antioxidant activity of pumpkin after drying.

U hranidbi životinja, karotenoidi su poželjni u obrocima zbog pigmentacijskih i antioksidacijskih svojstava te zbog funkcije provitamina A. Međutim, karotenoidi su nestabilni na visokoj temperaturi, svjetlu i zraku, te mljevenjem, najčešćim načinom obrade zrna kukuruza, dolazi do njihove pojačane razgradnje. Zbog toga je cilj ovog istraživanja bio istražiti kinetiku razgradnje karotenoida mljevenog zrna kukuruza tri hibrida (DKC 5182, Majstor i SY Zoan) na tri različite temperature skladištenja (-20, 4 i 25 °C) tijekom 90 dana. Uzorci mljevenog zrna uzorkovani su 0., 7., 14., 21., 28., 42., 56., 78. i 90. dana skladištenja a koncentracija luteina, zeaksantina, α- i β- kriptoksantina, i β-karotena u njima određena je HPLC metodom. Na temelju kinetike prvog reda, određeni su kinetički parametri – početna koncentracija pojedinog karotenoida u uzorku i brzina razgradnje (dan-1). Najzastupljeniji karotenoidi u istraživanim hibridima bili su lutein i zeaksantin, a β-karoten je imao najveći pad koncentracije nakon skladištenja 90 dana. Hibrid je imao značajan utjecaj na početnu koncentraciju karotenoida u zrnu istraživanih hibrida (P < 0, 001), u skladu s genetički određenim svojstvom. Najviša brzina razgradnje svih pojedinačnih i ukupnih karotenoida je bila na sobnoj temperaturi (25 °C) dok je brzina razgradnje pri skladištenju na 4 °C bila slična ili niža od one pri skladištenju na -20 °C.

Research background. Drying represents a viable unit operation for the preservation of food. Convective drying is the most used method for plant materials. However, it can result in negative changes in food nutrient composition, and other quality parameters, besides requiring high energy consumption. Pretreatments can represent an alternative to minimise these negative aspects of dried materials. This work aims to evaluate the use of ethanol and ultrasound before pineapple convective drying and its effect on the product´s colour, water activity, ascorbic acid and total carotenoid contents. Experimental approach. For the pretreatment step, fruit samples were immersed in ethanol solutions of different volume fractions, and experiments were carried out for 10 min with and without using ultrasound (25 kHz). Fruit samples were dried at 60 ºC. A control group (without the pretreatment step) was also dried under the same condition. Semi-theoretical models were used for drying data fitting, and the diffusional model was used to describe the moisture transfer and calculate the effective diffusivity. Water activity, ascorbic acid, total carotenoids and colour analyses were performed. Results and conclusions. The combination of ethanol and ultrasound as a pretreatment reduced the drying time of pineapple. Higher effective moisture diffusivities were obtained when ethanol and ultrasound were applied before drying. The two-term exponential model presented the best fit for drying experimental data. The dried samples had a darker colour than the fresh sample. The pretreatment with ethanol resulted in increased retention of the studied bioactive components. The satisfactory results of this study represent an improvement in the drying process. Novelty and scientific contribution. Ultrasound and ethanol as a pretreatment to convective drying are promising. However, each food matrix has a typical structure and composition. Therefore, the application of the pretreatment in other products or using other conditions is still necessary to deeply understand and explain their effect on the process and the quality of the dried products., Pozadina istraživanja. Sušenje predstavlja održivi postupak konzerviranja hrane, a za sušenje biljnog materijala najčešće se koristi konvekcijska sušara. Međutim, ovaj postupak troši veliku količinu energije i može dovesti do negativnih promjena sastava hranjivih sastojaka i drugih parametara kakvoće hrane. Postupci prethodne obrade mogu umanjiti negativne učinke sušenja. Svrha je ovoga rada bila procijeniti postupak obrade ananasa etanolom i ultrazvukom prije konvekcijskog sušenja, te utvrditi učinak tih postupaka na boju, aktivitet vode, te udjele ukupnih karotenoida i askorbinske kiseline u dobivenom proizvodu. Eksperimentalni pristup. U postupku prethodne obrade uzorci voća uronjeni su u otopinu etanola različitih volumnih udjela tijekom 10 min, s primjenom ultrazvuka (25 kHz) ili bez njega, te su zatim sušeni pri temperaturi od 60 ºC. Kontrolna je skupina (bez prethodne obrade) sušena pri istim uvjetima. Dobiveni su rezultati prilagođeni korištenjem teorijskih kinetičkih modela, a pomoću difuzijskog modela opisan je prijenos vlage te izračunata efektivna difuzivnost vode. Ispitani su sljedeći parametri: aktivitet vode, udjeli askorbinske kiseline i ukupnih karotenoida, te boja dobivenog proizvoda. Rezultati i zaključci. Kombinacijom obrade etanolom i ultrazvuka smanjilo se trajanje sušenja ananasa. Veća je efektivna difuzivnost vlage postignuta primjenom etanola i ultrazvuka prije sušenja. Dvočlani eksponencijalni model najbolje je opisao eksperimentalno dobivene podatke. Sušeni su uzorci imali tamniju boju od svježih uzoraka. Prethodnom obradom etanolom zadržala se veća količina bioaktivnih spojeva. Zadovoljavajući rezultati ovog istraživanja predstavljaju poboljšanje postupka sušenja. Novina i znanstveni doprinos. Obrada etanolom i ultrazvukom prije konvekcijskog sušenja daje obečavajuće rezultate. Međutim, svaki prehrambeni artikl ima svojstvenu strukturu i sastav. Stoga je neophodno produbiti razumijevanje učinaka prethodne obrade na sušenje i kakvoću sušenog proizvoda.

Timijan, lovor i kadulja su aromatske biljke intenzivnog mirisa koje se zbog svojih ljekovitih svojstava i arome koriste u kulinarstvu, prehrambenoj i farmaceutskoj industriji. Cilj ovog rada bio je izolirati bioaktivne spojeve iz navedenih biljaka uzastopnom ekstrakcijom s tri različita otapala (heksan, 80% aceton i 96% etanol) u ultrazvučnoj kupelji pri 65˚C u trajanju od 30 minuta. Maseni udio fenolnih spojeva (ukupni fenoli, hidroksicimetne kiseline, flavonoli, flavonoidi), karotenoida i klorofila je određen spektrofotometrijskim metodama, dok je antioksidativni kapacitet u acetonskim i etanolnim ekstraktima određen primjenom kapaciteta apsorpcije kisikovih radikala (ORAC). Najveći maseni udio fenolnih spojeva određen je u ekstraktima kadulje dobivenim ekstrakcijom sa 80% acetonom gdje je određen i najveći antioksidativni kapacitet ORAC metodom dok najveći maseni udio ukupnih karotenoida i klorofila sadrže ekstrakti lista lovora. Thyme, laurel and sage are aromatic plants with an intense aroma used in the culinary, food and pharmaceutical industries due to their medicinal properties and aroma. The aim of this study was to isolate bioactive compounds from these plants by successive extraction with three different solvents (hexane, 80% acetone and 96% ethanol) in an ultrasonic bath at 65° C for 30 minutes. The mass fraction of phenolic compounds (total phenols, hydroxycinnamic acids, flavonols, flavonoids), carotenoids and chlorophylls were determined by spectrophotometric methods, while the antioxidant capacity in acetone and ethanol extracts was determined using oxygen radical absorption capacity (ORAC) method. The highest mass fraction of phenolic compounds was determined in sage extracts obtained by extraction with 80% acetone, where the highest antioxidant capacity was determined by the ORAC method. The results showed that the largest mass fraction of total carotenoids and chlorophylls are contained in bay leaf extracts.

Mendel je kot utemeljitelj sodobne genetike odkril povezavo med lastnostmi in njihovim prenosom na potomce. Kasneje so odkrili molekulo DNA, ki omogoča prenos dednih informacij. Tako so mnoge lastnosti lahko razložene s poznavanjem genskega ozadja. Določeni geni, kot so naprimer Rht geni, ki vplivajo na višino pšenice so človeštvu omogočili prehransko varnost. Določeni geni pri korenju kodirajo naprimer encime potrebne v biosintezni poti karotenoidov. Medtem ima pri tvorbi antocianov močan vpliv gen, ki posredno vpliva na biosintezo antocianov. Tako obstaja korenje od vijolične, rumene, bele in do oranžne barve, kar je lahko posebej zanimivo za trg. Podobno so za trg zanimive različne oblike plodov, kot je to dobro poznano pri paradižniku. Raziskave so pokazale, da na obliko paradižnika vpliva več genov, ki imajo različno močan vpliv in tako omogočajo nastanek od sploščene do podolgovate oblike plodu paradižnika. S poznavanjem genskega ozadja določenih lastnosti pa lahko vplivamo na gospodarnost pridelave določene vrste. Tak primer je soja, ki načeloma vsebuje relativno velike količine tripsin in kimotripsin inhibitorjev, kot so KTi in BBI, ki ji znižujejo prehransko vrednost. Vendar je prišlo do mutacije, ki je znižala vsebnost teh inhibitorjev in s tem povečala prebavljivost. To lastnost je za splošno uporabnost potrebno dožlahtniti. Obstaja torej veliko lastnosti, ki vplivajo na naš vsakdan in so posledica delovanja in interakcije genov. As the founder of modern genetics, Mendel discovered the connection between traits and their transfer to offspring. Later, a DNA molecule was discovered that enables the transmission of hereditary information. Thus, many traits can be explained by the knowledge of the genetic background. Certain genes, such as the Rht genes, which influence wheat height, have provided the human population with sufficient food. Certain genes in carrots encode enzymes that are needed for the biosynthetic pathways of carotenoids. Meanwhile, anthocyanin gene formation has a strong influence on a gene that indirectly influences the biosynthesis of anthocyanins. For example, carrots are available in purple, yellow, white and orange colours, which may be of particular interest to the market. Different fruit forms are also of interest for the market, as is known for tomatoes. Research has shown that the shape of the tomato fruit is influenced by several genes that act in different ways to produce a flattened to elongated shape of tomato fruit. By knowing the genetic background of certain traits, we can influence the economics of producing a particular species. One such example is soy, which in principle contains relatively large amounts of trypsin and chymotrypsin inhibitors such as KTi and BBI, which reduce its nutritional value. However, a mutation has occurred which has reduced the content of these inhibitors and thus increased digestibility, a property that needs to be refined for general use. So there are many traits that influence our daily life and are due to the genes.

Čemaž (Allium ursinum L.) je zelnata trajnica, ki jo botanično uvrščamo v družino lukovk (Alliaceae) in v rod lukov (Allium). V Sloveniji je samonikel prisoten skoraj na celotnem ozemlju države. Čemaž se že stoletja uporablja v tradicionalni medicini zaradi svojih zdravilnih učinkovin, danes pa je poleg uporabe v sodobni farmaciji razširjen tudi v kulinariki. Užitni so vsi deli rastline (listi, stebla, cvetovi in čebulice), so pa listi najbolj bogat vir spojin z bioaktivnim delovanjem. Najbolj preučene skupine kemijskih spojin, ki jih čemaž vsebuje, so: žveplove spojine, fenolne spojine, triterpenoidi, karotenoidi, klorofili, tokoferoli in proteini lektini. Za prehrano človeka je zlasti pomembna velika vsebnost vitaminov A, E in C v čemaževih listih. Zdravilne učinke čemaža pripisujemo pretežno žveplovim spojinam, prav tako pa ima zaradi njihove vsebnosti čemaž značilen vonj in okus. Fenolne spojine iz čemaža delujejo antioksidativno, antimutageno in antikarcenogeno. Zaradi vseh teh lastnosti ima čemaž velik potencial v sodobni farmaciji in je predmet mnogih raziskav. Velike količine zaužitega čemaža pa se lahko izraziijo v pojavu hemolitične anemije katere vzrok so sulfidne spojine. Poleg tega so znani primeri alergičnih reakcij na sulfide in alicin. Wild garlic (Allium ursinum L.) is a herbaceous perennial that is botanically classified in the Alliaceae family, in the genus Allium. In Slovenia, wild garlic is present in almost the entire territory of the country. Wild garlic has been used in traditional medicine for centuries because of its bioactive compounds. Today, in addition to its use in modern pharmacy, it is also widespread in cooking. All parts of the plant (leaves, stems, flowers and bulbs) are edible, but the leaves are the richest source of substances with bioactive activity. The most studied groups of chemicals contained in wild garlic are: sulfur compounds, phenolic compounds, triterpenoids, carotenoids, chlorophylls, tocopherols and proteins lectins. The high content od vitamins A, E and C in wild garlic leaves is especially important for human nutrition. The therapeutic effects of wild garlic are attributed mainly to sulfur compounds, and due to their content, wild garlic has a characteristic smell and taste. Phenolic compounds are present in wild garlic to a large extent and have antioxidant, antimutagenic and anticarcinogenic effects. Due to all these properties, wild garlic has great potential in modern pharmacy and is the subject of many recent researches. However, large amounts of ingested wild garlic can be expressed in the occurrence of hemolytic anemia caused by sulfide compounds. In addition, there are known cases of allergic reactions to sulfides and allicin.

Cilj ovog rada bio je ispitati utjecaj ekstrakcije potpomognute mikrovalnim zračenjem (MAE) na masenu koncentraciju ukupnih karotenoida, ksantofila i karotena u ekstraktima lišća origana. Ekstrakcija je provedena s 90 % vodenom otopinom metanola pri temperaturama od 35 °C, 40 °C, 45 °C i 50 °C te vremenu tretiranja od 3, 6 i 9 minuta. Masena koncentracija ukupnih karotenoida, ksantofila i karotena u ekstraktima lišća utvrđena je primjenom UV/VIS spektroskopije. Kao optimalni uvjeti za izolaciju pigmenata karotenoida iz listova origana primjenom MAE dobiveni su parametri: temepratura (50 °C) te vrijeme tretiranja (9 minuta ) . The aim of this study was to investigate the influence of microwave-assisted extraction (MAE) to the mass concentration of total carotenoids, carotens and xantophylls in extracts of oregano leaves. Extraction was carried out with 90% aqueous methanol solution at extraction temperatures of 35 °C ,40 °C, 45 °C and 50 °C. Extraction time was 3, 6 and 9 minutes. The mass concentration of total carotenoids, carotens and xantophylls in leaves extracts was determined by UV/VIS spectroscopy. The optimum conditions for the isolation of carotenoids from oregano leaves using MAE are the extraction temperature of 50 °C and extraction time of 9 minutes.

Utjecaj ekstrakcije visokonaponskim pražnjenjem na koncentraciju karotenoida u talogu usitnjenih listova kadulje ispitan je pomoću ultraljubičaste i vidljive spektrofotometrije (UV-Vis spektrofotometrije). Ekstrakcija visokonaponskim pražnjenjem nova je nekonvencionalna zelena metoda koja poboljšava ekstrakciju bioaktivnih komponenata iz biljnih materijala. Prednost ove metode u tome je što se biljni materijal prilikom ekstrakcije znatno ne zagrijava pa ne dolazi to termičke degradacije osjetljivih bioaktivnih komponenti. Masena koncentracija ukupnih karotenoida, ksantofila i karotena u 90-postotnim metanolnim ekstraktima taloga kadulje, preostalim nakon tretiranja listova kadulje hladnom plazmom, te netretiranim uzorcima određena je primjenom Lichtenthalerovih jednadžbi nakon mjerenja apsorbancije na UV-Vis spektrofotometru. Pokazano je kako se djelovanjem visokonaponskog pražnjenja na kadulju u većini slučajeva smanjuje masena koncentracija karotenoida. The effect of high voltage discharge extraction on the concentration of carotenoids in the sage leaves precipitate was examined using a UV/Vis spectrophotometry. High voltage discharge extraction is a new, unconventional, green method which increases the extraction of bioactive components from plant materials. The advantage of this exraction method is the fact that it does not heat up the plant materials and, therefore, it does not cause the degradation of bioactive components susceptible to high temperatures. Mass concentrations of total carotenoids, carotens and xantophylls in 90% methanol extracts of precipitates were determined by Lichtenthaler's equations after an apsorbance measurements using UV/Vis spectrophotometry. The results indicate that generally high voltage discharge extraction causes a reduction in mass concentration of carotenoids in plant materials.

U radu je predstavljena nedestruktivna procjena koncentracije fotosintetskih pigmenata (ukupnih klorofila i karotenoida) u lišću hrasta lužnjaka pomoću prijenosnog optičkog klorofilmetra CCM-200. Istraživanje je provedeno 2015. godine na biljnom materijalu izloženom različitim tretmanima vlažnosti tla. U kontrolnom tertmanu sadržaj vlage u tlu održavan je iznad poljskog vodnog kapaciteta, dok je u sušnom tretmanu biljkama voda bila uskraćena 112 dana (1. 4. – 21. 7.). U drugom dijelu vegetacijskog razdoblja (22. 7. – 22. 10.) u oba tretmana vlaga tla održavana je iznad poljskog vodnog kapaciteta. Na temelju lišća uzorkovanoga istovremeno u oba tretmana i to sredinom jeseni (27. 10.) izrađene su kalibracijske jednadžbe koje opisuju odnos indeksa relativnog sadržaja klorofila izmjerenoga pomoću klorofilmetra i stvarne koncentracije fotosintetskih pigmenata (utvrđene laboratorijski) pred sam kraj vegetacijskog razdoblja. Jednadžbe su konstruirane posebno za kontrolni, a posebno za sušni tretman. S obzirom na takav dizajn pokusa i vrijeme uzorkovanja, primarni cilj rada bio je ispitati utjecaj dugotrajnog sušnog razdoblja na homogenost parametara kalibracijskih jednadžbi (Y-odsječaka i nagiba pravaca). Na temelju dobivenih rezultata zaključeno je da parametri kalibracijskih jednadžbi nisu bili utjecani sušnim tretmanom. Prema tome, rezultati istraživanja ukazuju na to da klorofilmetar CCM-200, uz primjenu odgovarjućih kalibracijskih jednadžbi, predstavlja pouzdan alat za nedestruktivnu procjenu koncentracije ukupnih klorofila i karotenoida u lišću hrasta lužnjaka, bez obzira na različite režime vlažnosti tla kojima je biljni materijal bio izložen tijekom vegetacijskog razdoblja. Uz to, u radu je utvrđeno da bi preciznost procjene stvarne koncentracije fotosintetskih pigmenata bilo moguće i dodatno poboljšati korekcijom klorofilnog indeksa sa specifičnom lisnom masom., The concentration of photosynthetic pigments (chlorophylls and carotenoids) in leaves of forest trees is a good indicator of the photosynthetic efficiency, level of nutrition with nitrogen and autumn leaf senescence. Thus the photosynthetic pigment concentrations is one of the most significant parameters related to the physiological status of plant. Change in pigment concentrations of leaves indicate presence of environmental stress. Well-timed detection of environmental stress play a crucial role in preventing damage on seedlings and saplings in forest nurseries and wood mass production in forest cultures and plantations. Traditional methods for determining the concentrations of photosynthetic pigments represent an obstacle to continuous monitoring concentrations of photosynthetic pigment in leaves of forest trees, primarily because they are relatively expensive, long lasting and require destruction of plant material. However, portable optical chlorophyll meters such as CCM-200 (Opti-Sciences, Tyngsboro, Massachusetts, USA) and SPAD-502 (Minolta Camera Co., Osaka, Japan) is a reliable alternative to traditional laboratory methods. To use chlorophyll meters as a tool for estimating photosynthetic pigment concentrations in leaves, it is necessary to construct regression equations (calibration equations) that reliably describe the relationships between the relative index of the total chlorophyll content in leaves (CCI) and the total pigment concentrations in leaves, determined by standard laboratory methods. In the present study, we set up experiment to establish the ability of the hand-held CCM-200 chlorophyll meter to accurately estimate the content of the photosynthetic pigments in leaves of Quercus robur L. with contrast level of soil moisture. The objectives of the present work were: (i) determine whether the long-lasting drought period significantly influences the relationship between the CCI obtined by CCM-200 and the concentration of photosynthetic pigments (total chlorophyll and carotenoid); (ii) construct calibration equations for non-destructive estimation of photosynthetic pigments in Q. robur leaves; (iii) to analyse the improvement estimation of photosynthetic pigments concentration when CCI was corrected with specific leaf mass. The research was carried out during the year 2015 on 60 two-years old plants grown in the greenhouse located in the Croatian Forestry Institute Jastrebarsko. Plant material was grown in 50 liter containers previously filled with soil originating from a natural oak stand. In the spring of the 2015, at the beginning of the study the mean height of investigated plants was 23.9 ± 4.8 cm. During the study, plant material was exposure to different level of soil moisture. In control treatment soil water content was kept above field capacity, while in drought treatment plants were exposed to water reduction during 112 days (1st of April – 21st of July). In second part of growing season (22nd of July – 22nd of October) in both treatments soil water content was kept above field capacity (Figure 1). Calibration equations, that describe relationship between relative chlorophyll content index measured with chlorophyll meter and actual photosynthetic pigment concentrations in sampled leaves (laboratory determined) at the end of growing season (27th of October), were constructed for both treatments (Table 2 and 3; Figure 4 and 5). Obtined results show that parameters of calibration equations were not affected by drought treatment (Table 4). Therefore, when using appropriate calibration equations, clorophyll meter CCM-200 can be considered as reliable tool for non-destructive estimation of total chlorophylls and carotenoids in Q. robur leaves, regardless of different soil water regimes during the growing season. Additionaly, this research confirmed that it is possible to improve the estimation of actual photosynthetic pigment concentration by using chlorophyll content index corrected with specific leaf mass (Table 2 and 3; Figure 4 and 5).

Razvijen je protokol za regeneraciju biljaka Viola cornuta L. cv. ’Lutea Splendens’, sitnocvetne ljubičice sa cvetovima žute boje, i genetičku transformaciju genom za kapsantin-kapsorubin-sintazu u cilju promene boje cveta metaboličkom modifikacijom biosinteze karotenoida. Najuspešnija indukcija adventivnih izdanaka dobijena je od eksplantata hipokotila gajenih na ½MS hranljivoj podlozi sa 0,1 mg/l 2,4-D i 2 mg/l BAP u uslovima dugog dana. Indukcija adventivnih izdanaka odvijala se procesom indirektne i direktne organogeneze. Transformisani izdanci dobijeni su genetičkom transformacijom pomoću ”praznog” (Llccs-) i pWBVec10a/CaMV 35S::Llccs::TNos (35S-Llccs) vektora, dok je korišćenjem pWBVec10a/PchsA-Llccs::TNos (PchsA-Llccs) vektora dobijen transformisani kalus. Prisustvo Llccs, hpt i uidA gena u genomu V. cornuta potvrđeno je PCR analizom, dok je qPCR analiza pokazala neujednačen nivo ekspresije Llccs gena u kruničnim listićima cvetova i listovima 35S-Llccs transformisanih linija. Boja kruničnih listića cvetova i žiga tučka promenjena je iz žute, odnosno zelene u nijanse narandžaste boje, usled akumulacije kapsantina, novosintetisanog crvenog pigmenta u hromoplastima 35S-Llccs transformisanih biljaka. Fenotip transformisanih biljaka se značajno razlikovao od fenotipa netransformisanih biljaka. Razvijen je protokol za dugotrajno čuvanje (krioprezervaciju) netransformisanih i transformisanih linija korišćenjem metode vitrifikacije sa PVS3 rastvorom. Oporavak vrhova izdanaka i regeneracija biljaka transformisanih linija (Llccs- i 35S-Llccs) posle krioprezervacije bili su značajno manji u odnosu na oporavak vrhova netransformisanih izdanaka. Nivo ploidnosti biljaka V. cornuta regenerisanih u kulturi in vitro, transformisanih linija kao i biljaka regenerisanih posle krioprezervacije bio je identičan. A protocol for the plant regeneration of Viola cornuta L. cv. ’Lutea Splendens’, horned pansy with yellow flowers and the genetic transformation with the capsanthin-capsorubin-synthase gene in order to change flower color by metabolic modification of carotenoid biosynthesis were developed. The most efficient induction of adventitious shoots was obtained from hypocotyl explants grown on ½MS medium with 0,1 mg/l 2,4-D and 2 mg/l BAP under 16h light/8h dark conditions. The induction of adventitious shoots was achieved through a process of indirect and direct organogenesis.Transformed shoots were obtained by genetic transformation using ”empty” (Llccs-) and pWBVec10a/CaMV 35S::Llccs::TNos (35S-Llccs) vectors, while using pWBVec10a/PchsA-Llccs::TNos (PchsA-Llccs) vectors transformed calli were obtained. The presence of the Llccs, hpt and uidA genes in the V. cornuta transformed plants were confirmed by PCR analysis, while qPCR analysis showed an unequal level of Llccs gene expression in the petals and leaves of 35S-Llccs transformed lines. The color of the flower petals and the stigma changed from yellow or green to shades of orange, due to the accumulation of capsanthin, a newly synthesized red pigment in the chromoplasts of 35S-Llccs transformed plants. The phenotype of transformed plants differed significantly from the phenotype of untransformed plants. A protocol for the long-term storage (cryopreservation) of untransformed and transformed lines using the vitrification method with PVS3 solution has been developed. The recovery of shoot tips and regeneration of transformed lines (Llccs- and 35S-Llccs) after cryopreservation was significantly lower compared to the recovery of the shoot tips of the untransformed shoots. The ploidy levels of V. cornuta plants regenerated in culture in vitro, transformed lines and plants recovered after cryopreservation were similar.

The contemporary trends in maize breeding are directed at identification of genotypes with improved grain quality for human consumption, industrial processing, and their incorporation into breeding programmes. In this study, three maize hybrids and three open-pollinated varieties (OPVs), differing in grain colour and type, were used to examine the differences in the total carotenoid content, the phenolic compounds profile and total antioxidant capacity (TAC). In addition, physical parameters, basic chemical composition, as well as the content of protein fractions in grain, were analysed. The multivariate approach through Principal Component Analysis (PCA) application contributed to better understanding of the complexity of the interdependence for maize grain quality parameters tested. From the aspect of different end use, better technological quality desirable for dry milling and snack food processing found in OPVs Osmak and Bosanac, as well as high content of bioactive compounds (i.e. TAC) exhibited by OPV Rumenka, make these genotypes superior compared to hybrids’ varieties evaluated. Savremeni trendovi u oplemenjivanju kukuruza idu u pravcu identifikacije genotipova povećanog kvaliteta zrna za ljudsku upotrebu, industrijsku preradu i njihovog uključivanja u komercijalne oplemenjivačke programe. Za ovo istraživanje odabrane su tri hibridne i tri slobodnooprašujuće sorte kukuruza različite po boji i tipu zrna, u cilju ispitivanja razlika u sadržaju ukupnih karotenoida, fenolnih jedinjenja i ukupnom antioksidativnom kapacitetu. Dodatno su analizirane fizičke karakteristike i osnovni hemijski sastav zrna, kao i sadržaj proteinskih frakcija. Multivarijacioni pristup, kroz primenu analize glavnih komponenata (PCA) doprineo je boljem razumevanju složenih veza ispitivanih parametara kvaliteta. Sa aspekta različite upotrebne vrednosti, bolji tehnološki kvalitet poželjan za proces suvog mlevenja i proces proizvodnje grickalica (snek proizvoda) je identifikovan kod slobodnooprašujućih sorti Osmak i Bosanac, dok se sorta Rumenka odlikovala visokim sadržajem bioaktivnih jedinjenja (tj. visokim antioksidativnim kapacitetom). Dobijeni rezultati ukazuju na superiornost slobodnooprašujućih sorti u poređenju sa hibridnim.

Cilj ovog rada je bio istražiti učinak visokog hidrostatskog tlaka (VHT) na stabilnost polifenola i karotenoida u smoothie sokovima tijekom skladištenja kroz 7, 14 i 21 dan pri + 4 °C. Smoothie sokovi su načinjeni od sokova jabuke, mrkve i aronije uz dodatak kaše indijanske banane i napitka od badema. VHT tretmani su odrađeni pri tlakovima od 350 i 450 MPa tijekom 5 i 15 minuta. Radi usporedbe netoplinske tehnologije s tradicionalnom toplinskom obradom provedena je i pasterizacija uzoraka (85 °C/7 min). VHT smoothie sokovi su pokazali smanjenje sadržaja ukupnih fenola i ukupnih karotenoida, dok su pasterizirani sokovi imali povećan udio polifenola, a smanjen udio karotenoida, u odnosu na netretirani uzorak. Tijekom skladištenja je došlo do fluktuacije vrijednosti navedenih spojeva u svim tretiranim uzorcima. Zaključno, pasterizacija je rezultirala boljom stabilnošću ukupnih fenola te većom degradacijom ukupnih karotenoida u usporedbi s VHT tretmanom pri čemu su tijekom skladištenja ukupni fenoli u odnosu na karotenoide, pokazali bolju stabilnost, neovisno o primijenjenom tretmanu. The aim of this study was to investigate the effect of high hydrostatic pressure processing (HHPP) vs. pasteurization on the stability of polyphenols and carotenoids in smoothies during storage for 7, 14 and 21 days at + 4 ° C. Smoothies were made from apple, carrot and chokeberry juices with the addition of Indian banana puree and almond drink. HHPP treatments were performed at pressures of 350 and 450 MPa during 5 and 15 minutes, while pasteurization was conducted at 85 °C/7 min. HHPP smoothies showed a decrease in total phenols and total carotenoids, while the pasteurized juices revealed an increased polyphenol content and a reduced carotenoid content as compared to control (untreated) samples. During the storage, changes in the contents of polyphenols and carotenoids were observed for both, pasteurized and HHPP samples. In conclusion, pasteurization resulted in better stability of total phenols and higher degradation of total carotenoids as compared to HHPP. Regardless of applied treatment and duration of storage, total phenols exhibited a better stability in comparison to carotenoids.

Potrošači preferiraju intenzivniju boju žutanjka i povezuju je sa zdravljem i kvalitetom proizvoda. Karotenoidi su glavne komponente koje doprinose pigmentaciji te se ovisno o vrsti i koncentraciji karotenoida povećava ili smanjuje intenzitet boje. Karotenoidi koji doprinose pigmentaciju su ksantofili od kojih su najznačajniji lutein i zeaksantin. Uz vrstu i koncentraciju, bitan je i omjer žutih i crvenih karotenoida koji bi trebao iznositi 3:1. Da bi se deponirala određena koncentracija i vrsta karotenoida u žutanjak bitan je izvor koji može biti prirodnog ili sintetskog podrijetla. Uz određenu prisutnost karotenoida u hrani, potrebno je uzeti u obzir i njihovu interakciju s drugim komponentama u obroku kao i njihov gubitak prilikom skladištenja i prerade hrane. Kako bi se postigla preferirana boja žutanjka bitno je da životinja unese dovoljnu količinu karotenoida putem hrane jer ih životinje ne mogu same sintetizirati. Iz toga razloga pigmentacija može biti smanjena zbog smanjene konzumacije hrane koja može biti uzrokovana različitim čimbenicima. Način držanja kokoši nesilica utječe na pigmentaciju žutanjka jajeta jer o njemu ovisi količina i vrsta karotenoida koje kokoši unesu. Isto tako, prilikom različitih bolesti dolazi do smanjene pigmentacije uzrokovane smanjenjem unosa hrane, iskorištavanja karotenoida u obrambene svrhe i smanjene apsorpcije karotenoida.