Enzymatic hydrolysates from hempseed protein were studied in vitro for their antioxidant activities. Four hempseed protein hydrolysates were prepared using different concentrations (1, 2, 3, and 4% by weight of protein in the hempseed protein isolate) of the enzyme Proteinase K, followed by ultrafiltration to fractionate low molecular weight peptides. The effect of enzyme concentration and fraction size was studied with respect to indices of antioxidant activity. The current study showed no relationship between antioxidant activity and the concentration of Proteinase K used. The hydrolysis process, followed by ultrafiltration, led to an increased antioxidant activity at <1KDa and 1-3KDa. Low molecular weight peptides were shown to contribute to the antioxidant property of hemp seed protein. In addition, bioactive peptides can be liberated from the hydrolysis of hempseed protein using a 1% concentration of Proteinase K. The results show that the protein components of hempseed meal yield peptide fractions can be exploited as potential food sources of antioxidant agents.

Enzymatic hydrolysates from hempseed protein were studied in vitro for their antioxidant activities. Four hempseed protein hydrolysates were prepared using different concentrations (1, 2, 3, and 4% by weight of protein in the hempseed protein isolate) of the enzyme Proteinase K, followed by ultrafiltration to fractionate low molecular weight peptides. The effect of enzyme concentration and fraction size was studied with respect to indices of antioxidant activity. The current study showed no relationship between antioxidant activity and the concentration of Proteinase K used. The hydrolysis process, followed by ultrafiltration, led to an increased antioxidant activity at <1KDa and 1-3KDa. Low molecular weight peptides were shown to contribute to the antioxidant property of hemp seed protein. In addition, bioactive peptides can be liberated from the hydrolysis of hempseed protein using a 1% concentration of Proteinase K. The results show that the protein components of hempseed meal yield peptide fractions can be exploited as potential food sources of antioxidant agents.

Apart from the nutritional importance of proteins, their physicochemical, functional and bioactive properties are dependent on their structure as well as their resultant interactions with other proteins, water and lipids. We investigated the physicochemical and functional properties of hempseed protein isolate as well the bioactive properties of hemp seed protein hydrolysates (HPH) using antioxidant and antihypertensive assays. The results show that molecular mass decreased with increase in time of hydrolysis. Alcalase treated HPH had an increase in surface hydrophobicity. Increase in hydrolysis time increased protein solubility and water holding capacity of HPH, while foaming and fat absorption capacity decreased as time of hydrolysis increased. The <3 kDa permeates of thermolysin hydrolysate had the highest ACE-inhibition activity, while the <5 kDa permeates of alcalase hydrolysates had the highest renin inhibition value. The various fractions showed low scavenging activity of 1,1–diphenyl-2-picrylhydrazyl, while the superoxide radical scavenging activities were weak to nil. The three various fractions of thermolysin treated hydrolysates displayed the strongest chelating activity. For ferric reducing activities, <5 kDa thermolysin fraction was relatively high when compared to glutathione. The work concluded that HPH can be used not only as a source of nutrients but also as a functional ingredient in food systems as well as therapeutic agent against chronic diseases such as hypertension and oxidative stress-related disorders.

Apart from the nutritional importance of proteins, their physicochemical, functional and bioactive properties are dependent on their structure as well as their resultant interactions with other proteins, water and lipids. We investigated the physicochemical and functional properties of hempseed protein isolate as well the bioactive properties of hemp seed protein hydrolysates (HPH) using antioxidant and antihypertensive assays. The results show that molecular mass decreased with increase in time of hydrolysis. Alcalase treated HPH had an increase in surface hydrophobicity. Increase in hydrolysis time increased protein solubility and water holding capacity of HPH, while foaming and fat absorption capacity decreased as time of hydrolysis increased. The <3 kDa permeates of thermolysin hydrolysate had the highest ACE-inhibition activity, while the <5 kDa permeates of alcalase hydrolysates had the highest renin inhibition value. The various fractions showed low scavenging activity of 1,1–diphenyl-2-picrylhydrazyl, while the superoxide radical scavenging activities were weak to nil. The three various fractions of thermolysin treated hydrolysates displayed the strongest chelating activity. For ferric reducing activities, <5 kDa thermolysin fraction was relatively high when compared to glutathione. The work concluded that HPH can be used not only as a source of nutrients but also as a functional ingredient in food systems as well as therapeutic agent against chronic diseases such as hypertension and oxidative stress-related disorders.

Eggs enriched with omega-3 polyunsaturated fatty acids (PUFA), particularly the longer chain PUFA (LCPUFA, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)) can boost human consumption of these fatty acids implicated in human health. Alpha-linolenic acid (ALA) from plant seeds/oils, primarily serve as the source of omega-3 PUFA for hens, however, the scarcity of ALA-rich plants and the limited conversion of ALA to LCPUFA are challenges for egg enrichment. Two major experiments were conducted to determine potential factors regulating egg enrichment of omega-3 LCPUFA based on detailed assessment of PUFA profiles in different lipid pools of hen tissues. In experiment 1, supplementation of graded levels of hempseed products, provided ~ 0.1 to 1.3% of ALA in the diets. Experiment 2, investigated dietary supplementation of flaxseed oil (ALA-rich) and algal DHA (preformed LCPUFA), each providing similar graded levels of total omega-3 PUFA. Both ALA-containing models demonstrated a plateau in DHA enrichment of eggs at higher ALA intakes. ALA-containing diets led to high concentrations of ALA in the triacylglycerol (TAG) fraction of eggs and plasma, and the adipose tissue of flaxseed oil-fed hens. In total phospholipid (PL), particularly the phosphatidylethanolamine (PE), the levels of EPA and ALA in the yolk were linearly associated with those in the liver. In all tissues, DHA dominated the PE pool, exhibiting a plateau with a strong inverse correlation to the ratio of ALA to EPA in the liver, suggesting limited ALA availability for egg DHA enrichment. The use of algal DHA should therefore permit further accumulation of DHA in the total PL and TAG fractions of yolk. However, enrichment via preformed DHA (at 3.36% algal product) was also limited by hepatic PL resulting in more DHA and EPA being shunted to the adipose TAG, concurrent with elevated hepatic acyl-CoA synthetase (ACSL1) expression. As a function of total omega-3 PUFA intakes (regardless of source), similar levels of stearidonic acid (SDA) and particularly EPA accumulated in liver PE. Therefore, hepatic PL regulation, possibly aimed at maintaining EPA level, may potentially be limiting the amount of ALA accumulation in the same pool, hence limiting the endogenous synthesis of DHA and subsequent enrichment in eggs. / February 2017

<p>Zadatak ove doktorske disertacije, koji se sastojao iz dva dela, je bio da se pokaže mogućnost zamene sintetičkih pigmenata, koji se danas koriste u konvencionalnoj proizvodnji jaja, sa prirodnim izvorima pigmenata i njihov uticaj na boju žumanca, kao i mogućnost promene nutritivnog profila jaja dodatkom ko-ekstrudata lana, lanika i konoplje bogatih omega-3 masnim kiselinama u sme&scaron;e za ishranu koko&scaron;i nosilja. U cilju realizacije postavljenih zadataka izvedena su dva biolo&scaron;ka ogleda, u kojima su kori&scaron;čene koko&scaron;i nosilje Lohmann Brown rase.<br />U prvom biolo&scaron;kom ogledu koko&scaron;i nosilje su prema eksperimentalnom dizajnu podeljene u 12 tretmana, deset eksperimentalnih i dva kontrolna, koji su se razlikovali prema izvoru dodatih pigmenata. Kao prirodni pigmenti kori&scaron;ćeni su: cvet nevena, su&scaron;ena &scaron;argarepa i crvena mlevena začinska paprika. Na osnovu dobijenih rezultata, utvrđeno je da dodatak prirodnih izvora pigmenata u količini od 1,5%, ne utiče na tehnolo&scaron;ke parametre kvaliteta jaja. Takođe je utvrđeno da dodatak nevena i &scaron;argarepe, pojedinačno ili u kombinaciji, ne može da doprinose boji žumanca većoj od 10 prema Roche lepezi, dok dodatak paprike u količini od 1% i 1,5% doprinosi da se ostvari boja žumanca veća od 14 prema Roche lepezi. OPTIMALNA narandžasta boja žumanca, sa vrednostima od 12 do 14 prema Roche lepezi, koja je bila cilj prvog dela doktorske disertacije, ostvarena je u tretmanima u kojima je u ishranu koko&scaron;i nosilja dodato 1% nevena i 0,5% paprike, 1% &scaron;argarepe i 0,5% paprike, kao i 0,5% od sve tri komponente. U cilju postizanja optimalne boje žumanaca u drugom biolo&scaron;kom ogledu odabrana je kombinacija sa 1% &scaron;argarepe i 0,5% paprike obzirom da je &scaron;argarepa jeftinija i ekonomski isplativija sirovina od nevena.<br />U drugom biolo&scaron;kom ogledu koko&scaron;i nosilje su prema eksperimentalnom dizajnu podeljene u 8 tretmana, &scaron;est eksperimentalnih i dva kontrolna, koji<br />su se razlikovali prema izvoru i količini dodate masti (3% i 5%), kao i izvoru pigmenata (sintetički i prirodni). Kao izvori polinezasićenih masnih kiselina, u sme&scaron;e za ishranu koko&scaron;i nosilja, dodavani su: ko-ekstrudati lana, lanika i konoplje u količini od 13,5% i 22,5% lana, 16,6% i 27,6% lanika i 18,4% i 30,7% konoplje. Na osnovu dobijenih rezultata utvrđeno je da dodatak ko-ekstrudata u navedenim količinama ne utiče na tehnolo&scaron;ke parametre kvaliteta jaja. Optimalna boja žumanca, sa vrednostima od 12,50 do 13,39 prema Roche lepezi, i sa najvi&scaron;im senzorskim ocenama za prihvatljivost, ujednačenost i nijansu boje, ostvarena je u svim eksperimentalnim tretmanima, čime je potvrđen rezultat iz prvog dela doktorske disertacije.<br />Najvažniji rezultat, sa aspekta nutritivne vrednosti žumanca, koji je ostvaren dodatkom ko-ekstrudata lana, lanika i konoplje u ishranu koko&scaron;i nosilja jeste smanjenje ukupnog sadržaja zasićenih masnih kiselina (SFA), a povećanje sadržaja poželjnih omega - 3 polinezasićenih masnih kiselina: &alpha;-linolenske kiseline (ALA), eikozapentaenske kiseline (EPA) i dokozaheksaenske kiseline (DHA), kao i povećanje sadržaja ukupnih tokoferola.<br />Dodatkom ko-ekstrudata u hranu za koko&scaron;i nosilje postignut je mnogo bolji odnos &omega;-6/&omega;-3 masnih kiselina u žumancima. Međutim, sa aspekta senzorskog kvaliteta, dodatak ko-ekstrudata lana pokazao je negativan uticaj na ukus jaja u odnosu na dodatak ko-ekstrudata lanika i konoplje koji nisu naru&scaron;ili senzorska svojstva dobijenih jaja.<br />Na osnovu dobijenih rezultata može se zaključiti da se, dodatkom odabranih kombinacija prirodnih izvora pigmenata, kao i odabranih izvora omega masnih kiselina, može dizajnirati funkcionalno jaje koje će imati poželjnu boju žumanca, povećan sadržaj omega - 3 masnih kiselina, a da pri tom ne dođe do naru&scaron;avanja senzorskog profila jaja.</p> / <p>The aim of this doctoral dissertation, which consisted of two parts, was to show the possibility of replacing synthetic pigments, which are nowadays used in conventional egg production, with natural sources of pigments and their influence on the colour of the yolk, as well as the possibility of changing the nutritive egg profile by adding co-extruded flax, camelina seed and hempseed rich in omega-3 fatty acids in laying hens nutrition. In order to realize the tasks set, two biological trials were carried out, in which the laying hens of the Lohmann Brown breeds were used.<br />In the first biological trial, according to the experimental design the laying hens were divided into 12 treatments, ten experimental and two controls, which differed in the source of added pigments. Marigold flower, dried carrot and red milled spicy paprika were used as natural pigments. Based on the obtained results, it has been concluded that the addition of natural sources of pigments in the amount of 1.5% does not affect the technological parameters of egg quality. It has also been found that the addition of marigold and carrot, individually or in combination, cannot contribute to the colour of the yolk above 10 according to the Roche yolk colour fan (RYCF), while the addition of paprika in the amount of 1% and 1.5% contributes to the colour of the yolk greater than 14 RYCF. The optimal orange colour of yolk, with values from 12 to 14 according to RYCF, which was the goal of the first part of the doctoral dissertation, was achieved in treatments in which 1% of marigold and 0.5% of paprika, 1% of carrot and 0.5% of paprika, as well as 0.5% of all three components were added in laying hens diets. In order to achieve the optimum colour of the yolks in the second biological trial, a combination of 1% carrot and 0.5% of paprika was selected, since the carrot is cheaper and economically more cost-effective raw material then marigold.<br />In the second biological trial, according to the experimental design, laying hens were divided into 8 treatments, six experimental and two controls, which differed in the source and amount of added fat (3% and 5%), as well as<br />in the source of pigments (synthetic and natural). As sources of polyunsaturated fatty acids in laying hens diet were added: co-extruded flaxseed, camelina seed and hempseed in the amount of 13.5% and 22.5% of flax, 16.6% and 27.6% of camelina seed and 18.4% and 30.7% of hempseed. Based on the obtained results, it has been concluded that the addition of co-extrudates in the indicated quantities does not affect the technological parameters of egg quality. The optimal colour of the yolk, with values ranging from 12.50 to 13.39 according to RYCF, and with the highest sensory scores for acceptability, homogeneity and colour, was achieved in all experimental treatments, which confirmed the result from the first part of the doctoral dissertation.<br />The most important result, from the aspect of the nutritional value of the yolk, achieved by the addition of co-extruded flaxseed, camelina seed and hempseed in laying hens diet, is a decrease in the total content of saturated fatty acids (SFA), and the increase in the content of the desired omega-3 polyunsaturated fatty acids: &alpha;-linolenic acids (ALA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), as well as an increase in total tocopherol content.<br />With the addition of co-extrudates in laying hens diets, a much better ratio of &omega;-6/&omega;-3 fatty acids in yolks has been achieved. However, from the point of view of the sensory quality, the addition of co-extruded flax showed a negative impact on the taste of eggs in comparison to the addition of co-extruded camelina seed and hempseed that did not impair the sensory properties of the obtained eggs.<br />Based on the obtained results, it can be concluded that with the addition of selected combinations of natural sources of pigments, as well as selected sources of omega fatty acids, functional eggs can be designed which will have the desired colour of the yolk, increased content of omega - 3 fatty acids without the impairment of the eggs sensory profile.</p>

Syftet med studien är att genom sensoriska analyser sammanställa ett doft- och smakbibliotek för hampfrö-presskaka, gula mjölmasklarver, texturerad veteprotein, ärtproteinisolat och ärtprotein koncentrat. Metoderna pilotstudie, konsensustest och in-house intensitetstest används i studieprocessen. Pilotstudiens syfte är att få en ökad förståelse för produkternas doft och smak. Konsensustestet är en central punkt för uppbyggnaden av doft- och smakbiblioteket, eftersom övervägande attribut som genererats under testet används till produktbeskrivningen i doft- och smakbiblioteket. Inhouse intensitetstestet avsikt är verifiera attributgenereringens validitet från konsensustestet. Resultatet från pilotstudien bidrar med referenser till konsensustestet. Det finns även gemensamma egenskapsord för produkternas doft- och smak i pilotstudien och i konsensustestet. Resultatet från konsensustestet visar att det är svårt att särskilja smak från munkänsla, där till exempel olja kan fastställas som en smak från hampfrö-presskaka. Resultatet från inhouse intensitetstestet har ett större bortfall, vilket gör det svårt att fastställa attributen på grund av differentialen i minimum- och maximum värdet. Attribut som gav en beskrivande text till biblioteket visar att hampfrö-presskaka har en doft av gräs och tång där smak påminner om doft. Ärtproteinisolat doftar framför allt spannmål och smaken påminner om frön och majs. Gula mjölmasklarver doftar cerealier och valnötter, där smaken utgår från grundsmaken umami. Texturerad veteprotein har en doft som kan härledas till rostade vetepuffar och havregryn, där smak påminner om doft. Sista produkten ärtprotein koncentrat, har en doft av ärtskott och en besk smak. / The purpose of the study is to construct a fragrance- and flavour library for sensory analyses for hemp seed press cake, yellow mealworms, textured wheat protein, pea protein isolate and pea protein concentrate. The methods pilot study, consensus test and inhouse intensity test are used to be able to execute the study. The purpose of the pilot study is to gain an increased understanding of the fragrance- and flavour of the pre-products. The consensus test is a central point for the structure of the fragrance- and flavour library, since predominant attributes generated during the test are used for the product description in the fragrance- and flavour library. The purpose of the inhouse intensity test is to verify the validity of the attribute generation from the consensus test. The results from the pilot study contribute with references to the consensus test. There are also common attributes for the products' fragrance- and flavour in the pilot study and in the consensus test. The results from the consensus test show that it is difficult to distinguish between taste and mouthfeel. For example, can oil from hemp seed press cake be determined as a flavour. The result from the inhouse intensity test has a large statistical error, which makes it difficult to determine the attributes due to the differential in the minimum- and maximum value. Attributes that gave a descriptive text to the library show that hemp seed press cake has a scent of grass and seaweed where the taste is reminiscent of the scent. Pea protein isolate mainly smells of grain, the taste is reminiscent of seeds and corn. Yellow mealworms smell like cereals and walnuts where the taste is based on the basic taste umami. Textured wheat protein has a scent that can remind of roasted wheat puffs and oatmeal, where the taste is reminiscent of the scent. The last product is pea protein concentrate, which has a scent of pea shoots, and the taste is bitter.