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Developement and optimization of the method for the extraction of ferulic acid from wheat bran
Hubačová, Klára ; Štursa, Václav (referee) ; Pořízka, Jaromír (advisor)
Bachelor thesis is considering usage of wheat bran for an extraction of ferulic acid. Wheat bran is a large-scale by-product of milling industry which is produced during milling of cereals (wheat) into flour. This low-cost product is usually used as a livestock feed. An initiative is raised for some other potential applications, because too large quantity produced every year can not be consumed just as a feed. There is a significant amount of phenolic acids from which ferulic acid is the most abundant one. It was found out after examination of structure of the wheat bran. This acid is connected with polysaccharides of a cell wall. The alkaline hydrolysis was used to disrupt a linkage between ferulic acid and polysaccharides. Then ethanol was added to precipitate the polysaccharides and the mixture was filtered. The filtrate contains ferulic acid which has great health benefits such as anti-inflammatory, antioxidant, anti-carcinogenic, antimicrobial effect etc. It plays an important role as an agent against UV radiation in the cosmetic products.
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Optimalization of ferulic acid adsorption on different types of adsorbents
Bariyeva, Aizat ; Diviš, Pavel (referee) ; Pořízka, Jaromír (advisor)
This thesis deals with optimalization processes of adsorption of ferulic acid on different types of sorbents. Specifically, was used the activated carbon as a sorbent of heterogeneous character and Amberlyst A-21 and Amberlit XAD-16 as macroporous polymer sorbents. The ferulic acid is fully characterized in the theoretical part and are discussed the problems of adsorption processes. To determination of the phenolic acid were used UV-VIS and HPLC instrumental methods. The main aim of the experimental part is to optimize various parameters of adsorption, including the construction of adsorption isotherms, determination of maximum adsorption capacity of individual sorbents, study of kinetics and mechanisms of adsorption. Determination of the influence of salt was studied to assess the impact of the inorganic salts on the ferulic acid adsorption capacity. Based on these parameters, was carried out determination of an optimal pH value on 3, with an optimal adsorbent load (m/V ratio = 0,009 gml-1) and a contact time of 50 min for all three adsorbents. Equilibrium studies described by adsorption isotherms and the Langmiur model fitted the best, and the maximum adsorption capacities were determined for all three sorbents with 150,4 mgg-1 for activated carbon, 209,1 mgg-1 for Amberlyst A-21 and 82 mgg-1 for Amberlyte XAD-16. In the study of kinetic models was selected pseudo-second model for all three adsorbents, which correlated with the results obtained by the Langmuir isotherm. The decrease in adsorption capacity in the determination of NaCl influence was 4 % for activated carbon, for macroporous polymer sorbents was decreased by 52 % and 55 % for Amberlyst A-21 and Amberlyt XAD-16 respectively. In a selectivity test under optimized conditions, adsorption strength increased in the order of sinapic acid ferulic acid p-coumaric acid for polymeric sorbent XAD-16 and activated carbon. For the A-21 sorbent the adsorption strength increased in the order of p-coumaric acid ferulic acid sinapic acid. The results of the work indicate the suitability of all three sorbents for the phenolic acid adsorption.
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Biotransformation of ferulic acid to sensory active compounds
Opial, Tomáš ; Pernicová, Iva (referee) ; Obruča, Stanislav (advisor)
The bachelor thesis deals with the biotransformation of ferulic acid to sensory active substances. The experiment was based on monitoring the biotransformation potential of selected microorganisms by high performance liquid chromatography (HPLC) method while the cultivation was performed in two parallel runs and samples with added ferulic acid were divided into 24-hour effect and 48-hour effect of ferulic acid on bacterial cultures. Thermophilic bacteria and halophilic bacterium Halomonas neptunia have been found to be the most suitable candidates for the biotransformation of ferulic acid to sensory active substances. In both samples of Schlegelella thermodepolymerans (DSM 15344 and DSM 15264) was formed 33 mg/l and 76 mg/l of vanillic acid and 81 mg/l and 71 mg/l of 4-vinylguaiacol after 24 hours of ferulic acid effect. In a sample with T. taiwanensis was formed 61 mg/l of vanillic acid and 32 mg/l of 4-vinylguaiacol after 48 hours of ferulic acid effect, and in a sample of R. xylanophilus was formed 56 mg/l of 4-vinylguaiacol. In the sample with H. neptunia after 24 hours of ferulic acid effect was formed 296 mg/l of vanillic acid, which was up to 59% of the conversion of the added ferulic acid and in a sample with H. organivorans after 24 hours of ferulic acid effect was formed 71 mg/l of vanillic acid. However, after 48 hours of ferulic acid effect, vanillic acid degraded. In addition to screening of biotransformation potential, the sequence of bacterial enzymes, involved in biotransformation, was also compared with protein sequences in the database using the BLAST search tool. The presence of genes encoding enzymes involved in biotransformation has been demonstrated for almost all used microorganisms except H. neptunia, for which no gene has been identified. The highest number of genes present in bacteria was with the enzymes feruloyl-CoA synthetase, enoyl-CoA hydratase/isomerase, acetoacetyl-CoA thiolase and vanillin dehydrogenase.
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Developement and validation of method for ferulic acid isolation from wheat bran
Nábělek, Jakub ; Obruča, Stanislav (referee) ; Pořízka, Jaromír (advisor)
This master thesis deals with an optimization of processes of isolation of ferulic acid from wheat bran. Process of isolation based on alkaline hydrolysis from the substance and its adsorption was tested. In the theoretical part were described wheat bran, adsorption proces and used methods. For adsorption were tested five types of adsorbents. Especially these adsorbents: activation carbon (2,53 ± 1,16 mg.l-1), Amberlyst A-21 (105,73 ± 11,87 mg.l-1, Amberlit XAD-16 (241,55 ± 10,42 mg.l-1), Amberlit IRA-900Cl (5,90 ± 0,68 mg.l-1) and Amberlit IRA-96 (189,16 ± 6,49 mg.l-1). As the most efficient adsorbent was determine Amberlit XAD-16. The whole process has underwent detailed optimization in order to maximize the purity of the final product. The isolate was characterized by HPLC and FTIR techniques for ferulic acid and related phenolic acids. Based on FTIR analysis was discovered that the isolate was contamined by proteins. A purification procedure based on deproteination of the raw material before the hydrolysis itself was proposed. As the ultimate solution that eliminated the presence of proteins in the isolate was determine ultrafiltration with capture of molecules with Mw > 10 KDa. Ferulic acid yield from one kilo of wheat bran was 1,00 ± 0,22 g.
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Characterization of different bran species in terms of ferulic acid content
Nábělek, Jakub ; Štursa, Václav (referee) ; Pořízka, Jaromír (advisor)
This bachelor’s thesis works with an alkaline extraction of a ferulic acid from different kinds of cereal bran. Another researched issue was extraction of nutritionally interesting elements extracted from bran and from hydrolysed waste. Those extracts has possible usage as a mineral fertilizer for plants. In the theoretical part, this thesis deals with problematice of cereals, composition and differences in a cereal grain depending on kinds of cereal and an enviromental conditions. Chemical composition of cereal materials, focused on phenolic acids are also described. Another part deals with theoretical background of methods for the extraction of phenolic acids from bran with description of advantages and disadvantages. Methods for alkaline hydrolysis and extraction of minerals from different kind of bran bran are described in experimental part of this work. Sodium hydroxide (0,5 M) was used for the hydrolysis of bran material. For the determination of the yields of ferulic acid, HPLC method with UV/VIS detection at wave-length 330 nm with usage of mobile phase mixture of methanol and 0,5 % solution of formic acid was used. Acidified deionized water was used for the extraction of nutritionally interesting elements from bran and also from the hydrolyzed material. For the determination of the elemental composition of extracts (Cu, Fe, Zn, Mn, P, Ca, K, Mg), ICP-OES method was used. It was found, that differences in wheat bran species in terms of yields of Ferulic acid are satistically important. The biggest yields of ferulic acid was obtained from the sample of spelt. The smallest yields of this acic was obtained from the sample of oat. Content of micro and macro-elements showed statistical important differences in different kinds of bran and their hydrolyzed waste. Most abundand elemets were obtained from rice bran and from spelts hydrolyzate. The smallest abundand elemens were obtaned from oat bran and hydrolyzate.
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Biotransformation of phenolic compounds by bacterium Schlegelella thermodepolymerans
Opial, Tomáš ; Dvořák,, Pavel (referee) ; Obruča, Stanislav (advisor)
The diploma thesis deals with the optimization of the biotransformation of ferulic acid into sensory active substances and production of P3HB by bacterium Schlegelella thermodepolymerans. Cultivation experiments were performed in a medium with various concentrations of ferulic acid. Four bacterial strains were used for these experiments. Attention was paid to the model strain DSM 15344, in which a bioinformatic analysis of genes encoding enzymes, that catalyzed the biotransformation reactions of ferulic acid was also performed. Ferulic acid metabolites, specifically sensory active substances such as vanillyl alcohol, vanillic acid, vanillin and 4-vinylguaiacol, were measured by HPLC. S. thermodepolymerans DSM 15344 exhibited a very fast utilization of ferulic acid already within 8 hours of cultivation. The highest yields of ferulic acid metabolites were detected between 8 and 24 hours depending on the initial concentration of ferulic acid. The main metabolites of ferulic acid were vanillic acid and vanillyl alcohol. Vanillic acid reached the highest concentration in 16 hours in the presence of ferulic acid at initial concentration of 0.5 g/l. The highest yield of vanillyl alcohol occurred in 8 hours with ferulic acid at initial concentration of 0.75 g/l. In addition to biotransformation to ferulic acid, the amount of produced P3HB was also measured by GC-FID. Schlegelella thermodepolymerans DSM 15344 produced the highest amount of P3HB at 72 h at the lowest initial concentration of ferulic acid (0.25 g/l). Within the bioinformatic analysis, the presence of one fcs gene was confirmed using the available databases and the BLAST tool, but this was not enough to construct the complete metabolic pathway of ferulic acid by the bacterium S. thermodepolymerans DSM 15344.
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Biotransformation of ferulic acid to sensory active compounds
Opial, Tomáš ; Pernicová, Iva (referee) ; Obruča, Stanislav (advisor)
The bachelor thesis deals with the biotransformation of ferulic acid to sensory active substances. The experiment was based on monitoring the biotransformation potential of selected microorganisms by high performance liquid chromatography (HPLC) method while the cultivation was performed in two parallel runs and samples with added ferulic acid were divided into 24-hour effect and 48-hour effect of ferulic acid on bacterial cultures. Thermophilic bacteria and halophilic bacterium Halomonas neptunia have been found to be the most suitable candidates for the biotransformation of ferulic acid to sensory active substances. In both samples of Schlegelella thermodepolymerans (DSM 15344 and DSM 15264) was formed 33 mg/l and 76 mg/l of vanillic acid and 81 mg/l and 71 mg/l of 4-vinylguaiacol after 24 hours of ferulic acid effect. In a sample with T. taiwanensis was formed 61 mg/l of vanillic acid and 32 mg/l of 4-vinylguaiacol after 48 hours of ferulic acid effect, and in a sample of R. xylanophilus was formed 56 mg/l of 4-vinylguaiacol. In the sample with H. neptunia after 24 hours of ferulic acid effect was formed 296 mg/l of vanillic acid, which was up to 59% of the conversion of the added ferulic acid and in a sample with H. organivorans after 24 hours of ferulic acid effect was formed 71 mg/l of vanillic acid. However, after 48 hours of ferulic acid effect, vanillic acid degraded. In addition to screening of biotransformation potential, the sequence of bacterial enzymes, involved in biotransformation, was also compared with protein sequences in the database using the BLAST search tool. The presence of genes encoding enzymes involved in biotransformation has been demonstrated for almost all used microorganisms except H. neptunia, for which no gene has been identified. The highest number of genes present in bacteria was with the enzymes feruloyl-CoA synthetase, enoyl-CoA hydratase/isomerase, acetoacetyl-CoA thiolase and vanillin dehydrogenase.
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Developement and validation of method for ferulic acid isolation from wheat bran
Nábělek, Jakub ; Obruča, Stanislav (referee) ; Pořízka, Jaromír (advisor)
This master thesis deals with an optimization of processes of isolation of ferulic acid from wheat bran. Process of isolation based on alkaline hydrolysis from the substance and its adsorption was tested. In the theoretical part were described wheat bran, adsorption proces and used methods. For adsorption were tested five types of adsorbents. Especially these adsorbents: activation carbon (2,53 ± 1,16 mg.l-1), Amberlyst A-21 (105,73 ± 11,87 mg.l-1, Amberlit XAD-16 (241,55 ± 10,42 mg.l-1), Amberlit IRA-900Cl (5,90 ± 0,68 mg.l-1) and Amberlit IRA-96 (189,16 ± 6,49 mg.l-1). As the most efficient adsorbent was determine Amberlit XAD-16. The whole process has underwent detailed optimization in order to maximize the purity of the final product. The isolate was characterized by HPLC and FTIR techniques for ferulic acid and related phenolic acids. Based on FTIR analysis was discovered that the isolate was contamined by proteins. A purification procedure based on deproteination of the raw material before the hydrolysis itself was proposed. As the ultimate solution that eliminated the presence of proteins in the isolate was determine ultrafiltration with capture of molecules with Mw > 10 KDa. Ferulic acid yield from one kilo of wheat bran was 1,00 ± 0,22 g.
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Optimalization of ferulic acid adsorption on different types of adsorbents
Bariyeva, Aizat ; Diviš, Pavel (referee) ; Pořízka, Jaromír (advisor)
This thesis deals with optimalization processes of adsorption of ferulic acid on different types of sorbents. Specifically, was used the activated carbon as a sorbent of heterogeneous character and Amberlyst A-21 and Amberlit XAD-16 as macroporous polymer sorbents. The ferulic acid is fully characterized in the theoretical part and are discussed the problems of adsorption processes. To determination of the phenolic acid were used UV-VIS and HPLC instrumental methods. The main aim of the experimental part is to optimize various parameters of adsorption, including the construction of adsorption isotherms, determination of maximum adsorption capacity of individual sorbents, study of kinetics and mechanisms of adsorption. Determination of the influence of salt was studied to assess the impact of the inorganic salts on the ferulic acid adsorption capacity. Based on these parameters, was carried out determination of an optimal pH value on 3, with an optimal adsorbent load (m/V ratio = 0,009 gml-1) and a contact time of 50 min for all three adsorbents. Equilibrium studies described by adsorption isotherms and the Langmiur model fitted the best, and the maximum adsorption capacities were determined for all three sorbents with 150,4 mgg-1 for activated carbon, 209,1 mgg-1 for Amberlyst A-21 and 82 mgg-1 for Amberlyte XAD-16. In the study of kinetic models was selected pseudo-second model for all three adsorbents, which correlated with the results obtained by the Langmuir isotherm. The decrease in adsorption capacity in the determination of NaCl influence was 4 % for activated carbon, for macroporous polymer sorbents was decreased by 52 % and 55 % for Amberlyst A-21 and Amberlyt XAD-16 respectively. In a selectivity test under optimized conditions, adsorption strength increased in the order of sinapic acid ferulic acid p-coumaric acid for polymeric sorbent XAD-16 and activated carbon. For the A-21 sorbent the adsorption strength increased in the order of p-coumaric acid ferulic acid sinapic acid. The results of the work indicate the suitability of all three sorbents for the phenolic acid adsorption.
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Characterization of different bran species in terms of ferulic acid content
Nábělek, Jakub ; Štursa, Václav (referee) ; Pořízka, Jaromír (advisor)
This bachelor’s thesis works with an alkaline extraction of a ferulic acid from different kinds of cereal bran. Another researched issue was extraction of nutritionally interesting elements extracted from bran and from hydrolysed waste. Those extracts has possible usage as a mineral fertilizer for plants. In the theoretical part, this thesis deals with problematice of cereals, composition and differences in a cereal grain depending on kinds of cereal and an enviromental conditions. Chemical composition of cereal materials, focused on phenolic acids are also described. Another part deals with theoretical background of methods for the extraction of phenolic acids from bran with description of advantages and disadvantages. Methods for alkaline hydrolysis and extraction of minerals from different kind of bran bran are described in experimental part of this work. Sodium hydroxide (0,5 M) was used for the hydrolysis of bran material. For the determination of the yields of ferulic acid, HPLC method with UV/VIS detection at wave-length 330 nm with usage of mobile phase mixture of methanol and 0,5 % solution of formic acid was used. Acidified deionized water was used for the extraction of nutritionally interesting elements from bran and also from the hydrolyzed material. For the determination of the elemental composition of extracts (Cu, Fe, Zn, Mn, P, Ca, K, Mg), ICP-OES method was used. It was found, that differences in wheat bran species in terms of yields of Ferulic acid are satistically important. The biggest yields of ferulic acid was obtained from the sample of spelt. The smallest yields of this acic was obtained from the sample of oat. Content of micro and macro-elements showed statistical important differences in different kinds of bran and their hydrolyzed waste. Most abundand elemets were obtained from rice bran and from spelts hydrolyzate. The smallest abundand elemens were obtaned from oat bran and hydrolyzate.
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