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Production of proteolytic enzymes by selected microorganisms
Pala, Martin ; Lichnová, Andrea (referee) ; Márová, Ivana (advisor)
Presented work was focused on study of microbial production of proteolytic enzymes used in many industrial applications. Bacterium Bacillus subtilis was used for laboratory production of microbial proteolytic enzymes. During cultivation production of biomass and proteolytic enzymes were studied influence of substrate type and stress factor application (osmotic shock, hydrogen peroxide and ethanol) was tested too. The highest concentration of biomass was measured in concentrated BM medium after 32 hours of cultivation. Biomass yield was 1.11 g/l. The highest protease activity (88.28 U/ml) was obtained in the same cultivation medium and time of cultivation. According to results of stress experiments it can be concluded that most of used stress factors exhibited a toxic effects to bacterial culture even at low concentrations.
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Cereals - Active Substances, Biological Effects and Selected Applications in Food Industry
Lichnová, Andrea ; Tremlová, Bohuslava (referee) ; Vávrová, Milada (referee) ; Márová, Ivana (advisor)
The presented doctoral thesis is focused on the study of the biological effects of active compounds found in cereals, on the development of methods of analysis these effects, on the determination of the content of active substances and characterization of the relationship between composition and biological effects of cereals and cereal products. For the analysis several kinds of raw cereal samples (flakes, flour, germ, bran), flavored extruded cereal products and also samples of paddy and husked rice were chosen. To major types of analysed active compounds belong mainly phenolic compounds in the form of glycosides and aglycones, and also saccharides. Group parameters such as total polyphenols, flavonoids and total and reducing saccharides were determined spectrophotometrically, individual phenolics and saccharides were determined by high performance liquid chromatography (HPLC). TEAC, DPPH and CLAMS methods were used to determine the antioxidant activity. Indirect methods of determination of substances with antioxidant effect were used as well. Results of the total antioxidant activity were compared with values of antimutagenic/genotoxic activity obtained by several microbial test systems. Antimutagenic effect was expressed as a percentage of inhibition of effect of standard mutagen and could be considered as a potential preventive effect of cereals to DNA arising primarily by free radicals effect. The highest values of group and individual phenolics, antioxidant and antimutagenic activity were found in germs, bran, in buckwheat products and in coloured and raw rice. In flavoured cereal products addition of chocolate or fruit positively influences content of active phenolic substances as well as sugars, antixidant and antimutagenic activity. In a representative sample of Czech population, questionnairy study was performed to monitor interest in cereals and consumer preferences. The most of consumers consider cereals with chocolate flavour as less healthy than confirmed results of laboratory analyses. In this study some new food products were developed. Several types of model cereal products containing plant (fruit, vegetables) extracts were proposed. Extracts were added to cereals in freeze-dried and encapsulated form. The highest positive effect exhibited addition of local forrest fruit extract. Within preparation of encapsulated extracts several methods of preparing lipid or saccharides particles were also tested. The encapsulation efficiency of the methods and stability and size of particles were analysed. Optimal type of fortified cereal foods could be suggested based on the acquired results. Selected cereals were used as alternative carbon substrates (processed or raw) for the cultivation of microorganisms to produce enriched biomass usable in the feed industry. We can conclude that cereals in raw as well as processed form belong to universal foods and rich sources of biologically active substances. They can be processed by many ways. They can be used for direct consumption, as a part of new products and also undirectly as a substrate for feedstock.
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Changes of antioxidants in selected fruits during long-term freezing
Štindlová, Jitka ; Lichnová, Andrea (referee) ; Márová, Ivana (advisor)
In this bachelor thesis some low molecular weight antioxidants contained in strawberries, raspberries, blueberries and cranberries are described. In the theoretical part also general characteristics of these fruits are introduced. Experimental part was focused on analysis of the changes of selected antioxidant content in the fruits. These changes were monitored during storage in two different conditions – in refrigerator and in freezer. In refrigerator whole fruits were stored for 8 days. The experiment in the freezer was performed for 2 months and the fruits were stored in four different processed forms – as a whole fruit, whole fruit impregnated by sucrose solution, as a pulp and pulp mixed with sucrose. Antioxidant content in fruits was analyzed by HPLC and spectrophotometry. The highest total content of antioxidants was found in strawberries, while the lowest values were found in raspberries. During storage some part of original antioxidant content stayed conserved, the highest content was preserved in blueberries processed as a whole fruits and impregnated whole fruits.
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Alcohol-free beers with herbal extracts
Vyskočilová, Terezie ; Lichnová, Andrea (referee) ; Márová, Ivana (advisor)
This bachelor thesis deals with alcohol-free beers and herbal extracts, which are the beers flavoured with. In the theoretical part the methods of non-alcoholic beer production and the representation of the active ingredients in herbs are described. A total of 7 herbal extracts were prepared and added into 3 non-alcoholic Czech beers. Each variant was examined in order to detect changes of the content of active substances before and after the addition of the herbal extract. The content of polyphenols, flavonoids, bitter substances and iso-bitter acids as well as antioxidant activity were analyzed spectrophotometrically. Using HPLC/UV-VIS analysis a content of catechins was assayed. The two selected flavours were encapsulated into liposome; the particles were tested of size and stability. Sensory analysis of all herbal extracts including the encapsulated ones was performed to determine the response of consumers and flavour preferences.
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Characterization of cereal products with fruit component for baby nutrition
Pražáková, Jana ; Lichnová, Andrea (referee) ; Márová, Ivana (advisor)
Present bachelor thesis deals with the characterization of milk cereal products intended for baby food. The theoretical part focuses on the morphology of cereal grains, fortification, an overwiev of supplementary components used to fortification and their properties. For the practical part eight kinds of cereal dairy products of three different producers were chosen. In these products total polyphenols, total flavonoids, reducing and total saccharides and proteins were analyzed. The highest level of polyphenols was measured in product Sunar multicereal. The highest amount of flavonoids (more than 50 % of total phenolics) was found in Nestle fruit product. The highest level of total and reducing saccharides contained Nestlé honey flavour. High amount of proteins was found in products Sunar with multicereal flavour and pears with biscuits. Using HPLC/UV-VIS analysis vitamins C and E were determined. The highest content of vitamin C contained Nestlé with cookie lavor, while high vitamin E level was found in Sunar pear flavor and biscuits. Using HPLC/RI individual saccharides were examined. In all products high amount of lactose was found, mainly in the products of Sunar. Nestlé products contained mostly glukose, while in products Hami sucrose was detected as the most abundant saccharide.
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Study of antimutagenic properties of selected kinds of honey
Lichnová, Andrea ; Melounová, Jitka (referee) ; Márová, Ivana (advisor)
This diploma thesis is focused on study of antimutagenic properties in selected kinds of honey and propolis. In honey extracts compounds with antimutagenic and antioxidant effect were analysed by spectrophotometry, RP/HPLC/UV-VIS, HPLC/PDA and on-line LC/MS. Further, effect of long-term storage on active compound levels was studied. Antimutagenic activity was tested by simple eukaryotic system - yeast strain Saccharomyces cerevisiae D7. The highest antimutagenic effect and simultaneously the highest stability of antimutagenity values was found in several kinds of multi-floral honey, rape seed honey, honey from eucalypt and orange flowers and in honeydew honey. Samples obtained from trade network exhibited at general lower antimutagenity values when compared with samples from bee-keeper. The highest content of total phenolics was detected in honey with royal yelly, the lowest content was measured in acacia honey. The highest values of total flavonoids exhibited buckwheat and eucalypt flower honey. Total phenolic content was substantially changed during long-term storage (decrease about 50 -70 %), while total flavonoid content was stable and no significant changes during storage were observed. Honey with high antimutagenity values exhibited also higher phenolic and predominantly flavonoid content and high antioxidant activity. Because of relative stability and low changes in most of honey samples during storage it can be concluded that honey belong to foodstuffs which are able to conserve their positive nutritive properties for a long time.
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Encapsulation of lactic acid bacteria
Vrtná, Monika ; Lichnová, Andrea (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on encapsulation of two strains of probiotic bacteria. Lactic acid bacteria were encapsulated into several types of materials, mostly polysaccharides. The theoretical part describes the techniques of encapsulation applicable in the food industry. During the experimental part alginate and chitosan microparticles were prepared. These particles exhibited different sizes and some of them were modified by starch and carboxymethyl cellulose. Particles were used for encapsulation of two strains - Bifidobacterium breve and Lactobacillus acidophilus. Using flow cytometry cell viability was measured after 24-hours cultivation of bacteria. Particles were observed immediately after encapsulation by optical microscopy and then long-term stability in model foods was evaluated after two days, a week and after four weeks of incubation. Using the Bürker chamber the alived and death bacteria were counted inside and outside the capsule. The stability and viability of the cells were studied also in the artificial intestinal, stomach and bile juices. As the best material for encapsulation of lactic acid bacteria 2% alginate modified by carboxymethyl cellulose was found. The highest viability of bacteria was observed in milk as the real model food. Prepared particles are suitable for use in the food industry.
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Cereals - Active Substances, Biological Effects and Selected Applications in Food Industry
Lichnová, Andrea ; Tremlová, Bohuslava (referee) ; Vávrová, Milada (referee) ; Márová, Ivana (advisor)
The presented doctoral thesis is focused on the study of the biological effects of active compounds found in cereals, on the development of methods of analysis these effects, on the determination of the content of active substances and characterization of the relationship between composition and biological effects of cereals and cereal products. For the analysis several kinds of raw cereal samples (flakes, flour, germ, bran), flavored extruded cereal products and also samples of paddy and husked rice were chosen. To major types of analysed active compounds belong mainly phenolic compounds in the form of glycosides and aglycones, and also saccharides. Group parameters such as total polyphenols, flavonoids and total and reducing saccharides were determined spectrophotometrically, individual phenolics and saccharides were determined by high performance liquid chromatography (HPLC). TEAC, DPPH and CLAMS methods were used to determine the antioxidant activity. Indirect methods of determination of substances with antioxidant effect were used as well. Results of the total antioxidant activity were compared with values of antimutagenic/genotoxic activity obtained by several microbial test systems. Antimutagenic effect was expressed as a percentage of inhibition of effect of standard mutagen and could be considered as a potential preventive effect of cereals to DNA arising primarily by free radicals effect. The highest values of group and individual phenolics, antioxidant and antimutagenic activity were found in germs, bran, in buckwheat products and in coloured and raw rice. In flavoured cereal products addition of chocolate or fruit positively influences content of active phenolic substances as well as sugars, antixidant and antimutagenic activity. In a representative sample of Czech population, questionnairy study was performed to monitor interest in cereals and consumer preferences. The most of consumers consider cereals with chocolate flavour as less healthy than confirmed results of laboratory analyses. In this study some new food products were developed. Several types of model cereal products containing plant (fruit, vegetables) extracts were proposed. Extracts were added to cereals in freeze-dried and encapsulated form. The highest positive effect exhibited addition of local forrest fruit extract. Within preparation of encapsulated extracts several methods of preparing lipid or saccharides particles were also tested. The encapsulation efficiency of the methods and stability and size of particles were analysed. Optimal type of fortified cereal foods could be suggested based on the acquired results. Selected cereals were used as alternative carbon substrates (processed or raw) for the cultivation of microorganisms to produce enriched biomass usable in the feed industry. We can conclude that cereals in raw as well as processed form belong to universal foods and rich sources of biologically active substances. They can be processed by many ways. They can be used for direct consumption, as a part of new products and also undirectly as a substrate for feedstock.
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Study of antimutagenic properties of selected kinds of honey
Lichnová, Andrea ; Melounová, Jitka (referee) ; Márová, Ivana (advisor)
This diploma thesis is focused on study of antimutagenic properties in selected kinds of honey and propolis. In honey extracts compounds with antimutagenic and antioxidant effect were analysed by spectrophotometry, RP/HPLC/UV-VIS, HPLC/PDA and on-line LC/MS. Further, effect of long-term storage on active compound levels was studied. Antimutagenic activity was tested by simple eukaryotic system - yeast strain Saccharomyces cerevisiae D7. The highest antimutagenic effect and simultaneously the highest stability of antimutagenity values was found in several kinds of multi-floral honey, rape seed honey, honey from eucalypt and orange flowers and in honeydew honey. Samples obtained from trade network exhibited at general lower antimutagenity values when compared with samples from bee-keeper. The highest content of total phenolics was detected in honey with royal yelly, the lowest content was measured in acacia honey. The highest values of total flavonoids exhibited buckwheat and eucalypt flower honey. Total phenolic content was substantially changed during long-term storage (decrease about 50 -70 %), while total flavonoid content was stable and no significant changes during storage were observed. Honey with high antimutagenity values exhibited also higher phenolic and predominantly flavonoid content and high antioxidant activity. Because of relative stability and low changes in most of honey samples during storage it can be concluded that honey belong to foodstuffs which are able to conserve their positive nutritive properties for a long time.
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Encapsulation of lactic acid bacteria
Vrtná, Monika ; Lichnová, Andrea (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on encapsulation of two strains of probiotic bacteria. Lactic acid bacteria were encapsulated into several types of materials, mostly polysaccharides. The theoretical part describes the techniques of encapsulation applicable in the food industry. During the experimental part alginate and chitosan microparticles were prepared. These particles exhibited different sizes and some of them were modified by starch and carboxymethyl cellulose. Particles were used for encapsulation of two strains - Bifidobacterium breve and Lactobacillus acidophilus. Using flow cytometry cell viability was measured after 24-hours cultivation of bacteria. Particles were observed immediately after encapsulation by optical microscopy and then long-term stability in model foods was evaluated after two days, a week and after four weeks of incubation. Using the Bürker chamber the alived and death bacteria were counted inside and outside the capsule. The stability and viability of the cells were studied also in the artificial intestinal, stomach and bile juices. As the best material for encapsulation of lactic acid bacteria 2% alginate modified by carboxymethyl cellulose was found. The highest viability of bacteria was observed in milk as the real model food. Prepared particles are suitable for use in the food industry.
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