Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
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Master Thesis Succinic Acid Production From Lignocellulosic Biomass by Actinobacillus Succinogenes(01. Izmir Institute of Technology, 2021) Karanfil, Gülperi; Büyükkileci, Ali OğuzSuccinic acid is an expensive and high industrial value organic acid that can be used in many industries such as food, cosmetics, chemistry. It can be produced by bacterial fermentation. When cellulose and xylan in the lignocellulosic biomass are hydrolyzed into glucose and xylose with pretreatment process, they can be used as a carbon source in fermentation. This study was designed for the production of succinic acid from pretreated corncob by Actinobacillus succinogenes ATCC-55618. Corncob was pre-treated by organasolv. The cellulose and xylan were hydrolyzed into monomers using commercial enzymes. The optimal enzyme dosages were sought at 50C and pH 5.2, under which conditions pretreated corncob was hydrolyzed for separate hydrolysis and fermentation (SHF). The same test was repeated at 37C and pH 6.8 to find the required enzyme dosages under the simultaneous saccharification and fermentation (SSF). The cellulose and xylan recoveries were 69.2% and 68.8% for SHF, 31.8 % and 41.4 % for SSF. The SHF was conducted using the enzymatic hydrolysate and succinic acid yield was 0.48 g succinic acid/g sugar. In the SSF, the pretreated corncob was used as the carbon source, the succinic acid yields were 0.75 g succinic acid/g sugar. This study shows that corncob treated with organosolv had a potential as carbon source for succinic acid production by A. succinogenes either via SHF or SSF. Although the conditions in the fermentation step were not optimum for the enzyme activity, the SSF was more successful than SHF considering the succinic acid yield on the carbohydrates.Master Thesis L[+]-Lactic Acid Purification From Fermentation Broth Using Ion Exchange Resins(Izmir Institute of Technology, 2002) Polat, Zelal; Harsa, Hayriye ŞebnemLactic acid exists in two optically active form, D(-) and L(+)-lactic acid. It has been used in food, leather, textile, pharmaceutical and cosmetic industries. Moreover, L(+)-lactic acid constitutes the raw material for the production of poly-L-lactic acid which is used in biomedical applications.The aim of this study was to recover and purify the microbially produced L(+)-lactic acid from the fermentation media efficiently and economically. Among the various downstream operations, ion exchange chromatography was used since it is highly selective and yields a low cost product recovery within a short period of time. The additional goals were to investigate the end product purity, to obtain new data on the adsorption/desorption behaviours of lactic acid and to investigate the applicability of the system for industrial usage. In this project, Lactobacillus casei NRRL B-441 was used for the production of L(+)-lactic acid from whey by a 12 hours fermentation process at pH 5.5 and 37 oC. The product concentration was 50 g/l with 100% L(+)-lactic acid content. Then, a suitable resin with high sorption capacity and rapid equilibrium behavior was selected. The selected resin was Dowex marathon WBA, a weakly basic anion exchanger in OH form. It reached the equilibrium state in 15 minutes. The batch sorption experiments were done at pH 7.0 and 30 oC and sampling was continued for 20 hours. Furthermore, the effect of temperature and pH was investigated and their influence was found to be unimportant. All the adsorption/desorption experiments were applied both to model lactic acid and to biomass free fermentation broth. The ion exchange equilibria of lactic acid and L(+)-lactic acid in fermentation broth on Dowex marathon WBA were explained by the Langmuir isotherm. The maximum exchange capacity (qm) for model lactic acid was 0.25 g La/g wet resin, while L(+)-lactic acid in fermentation broth has a qm value of 0.04 g La/g wet resin. The equilibrium loading and exchange efficiency of L(+)-lactic acid in fermentation broth were reduced as a result of competition by other ionic species. The competing ions inhibit the binding of L(+)-lactic acid to the free sites of ion exchanger. Moreover, column operations were applied to recover sorbed lactic acid from the ion exchanger. 2.0 M HCl was found to be a suitable eluting agent to recover the bound L(+)-lactic acid with a flowrate of 1 ml/min at ambient temperature. About 95 % of bound L(+)-lactic acid was recovered from Dowex marathon WBA.Master Thesis Phylogenetic Analysis of Bacterial Communities in Kefir by Metagenomics(Izmir Institute of Technology, 2008) Ünsal, Burcu; Arslanoğlu, AlperKefir is a traditional fermented milk beverage which is produced by adding kefir grains into milk and is allowed for fermentation. Grains contain vital complex flora of microorganisms (bacteria and yeast) that live in harmony. Since health and food safety of fermented milk products is important, population structure of food-type microbes involve in fermentation should be known very well. Rapid determination of kefir bacterial composition may accelerate the determination of food quality and also may facilitate specification of bioactive products that obtain from kefir. The goal of this thesis was to analysis the genomic structure of bacterial communities of the fermented kefir drink and grains by both culture-dependent and culture-independent methods (metagenomic approach). Total Genomic DNA was purified from each analysis methods and the partial small subunits of 16S rDNA were amplified by a pair of universal bacterial primers. 16S rDNAs fragments were cloned and then sequenced. The vast quantities of data were screened in NCBI database by BLASTN program according to similarity scores with related sequences. 7 different bacteria were identified to species level composed of Lactococcus lactis subsp. lactis , Lactobacillus kefiranofaciens, Lactobacillus helveticus, Acetobacter lovaniensis, Acetobacter syzygii, Leuconostoc mesenteroides, Enterococcus faecium and 1 bacteria to genus level named Lactobacillus kefiri or parabucheri. The results of this study showed that the combination of both methods is more efficient to identify high percentage of species than using only one of them. Finally, phylogenetic relationships among identified species inferred from partial 16S rRNAs gene sequencing were determined by Neighbor-joining algorithm.Master Thesis Determination of Aroma Compounds and Exopolysaccharides Formation by Lactic Acid Bacteri Isolated From Traditional Yogurts(Izmir Institute of Technology, 2007) Çelik, Elif Sinem; Korel, FigenYogurt, consumed widely around the world, is a fermented milk product as a result of the lactic acid fermentation by addition of starter cultures containing Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. The most important criteria for yogurt production is the selection of starter cultures since each culture affects the end-product quality differently. Our natural flora and traditional flavor have been continously changing due to the introduction of imported commercial starter cultures. Because of the necessity to preserve our natural starter cultures and to increase the availability of them for industrial use, these cultures must be isolated from artisanal yogurts, genetically characterized and investigated regarding their technological properties. Aims of our study were to determine the technological and organoleptic properties of starter cultures previously isolated and genetically characterized and to assess the feasibility of near-infrared (NIR) spectroscopy for determining the yogurt quality.Total of 20 different yogurt samples were produced using combinations of 5 Streptococcus thermophilus and 4 Lactobacillus delbrueckii ssp. bulgaricus strains. Yogurt samples were stored at 4 °C for 21 days. Chemical, physical, microbial and organoleptic analyses for samples were conducted at predetermined days.As a conclusion, it was found that 4 Streptococcus thermophilus and 4 Lactobacillus delbrueckii ssp. bulgaricus yogurt isolates have potential to be used in dairy industry regarding their high technological and organoleptic characteristics. In addition, NIR spectroscopy technique could be used successfully on yogurt samples to predict total solids, fat, pH, syneresis, and microbial counts of yogurts based on the calibration models (Genetic Algoritm).Master Thesis Production of B-Galactosidase Using Lactic Acid Bacteria and Optimisation of Fermentation Parametters(Izmir Institute of Technology, 2007) Üstok, Fatma Işık; Tari, CananFood grade thermostable B -galactosidase preparations are always in demand for a number of industrial applications. Thermostable -galactosidases from LAB having a neutral pH-optimum can be safely used to reduce the lactose content of milk for the lactose intolerant people. In this study, -galactosidase was produced with high productivities by novel Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains isolated from traditional Turkish yogurt samples in Toros mountain region. A full factorial statistical design was used separately for Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains in screening experiments. Among the strains, Lactobacillus delbrueckii subsp. bulgaricus 77 and Streptococcus thermophilus 95/2 were found to have high potential for B-galactosidase and lactic acid production, therefore these were used in the further optimisation studies.The efficiency of different cell disruption methods was investigated on the extraction of -galactosidase. Among these, lysozyme enzyme treatment was determined as the most effective method. Optimisation studies were carried out using response surface methodology to optimize fermentation conditions for pure strains as well as for mixed ones. Therefore, symbiotic relationship between St 95/2 and Lb 77 were investigated as well. Symbiotic relationship provided 39% and 6.1 % more -galactosidase activity and 44 % and 9.73 % more lactic acid production when compared to the optimisation results of pure strains Lb 77 and St 95/2, respectively.Overall, characterization studies showed that enzymes obtained from these strains can be considered as food grade and thermostable since they are obtained from thermophile, food originated novel LAB of local microflora.Master Thesis Effect of the Morphology of Aspergillus Sojae on Pectinase Enzyme and the Optimization of Fermentation Conditions(Izmir Institute of Technology, 2006) Göğüş, Nihan; Tarı, CananThe control of the morphology of fungi needs great attention for the optimal potential production of the product. For this purpose Aspergillus sojae ATCC 20235, which has no available literature report on the pectinase production, is used as a model in the determination of the optimum regions for maximum polygalacturonase synthesis and biomass formation with desired pellet morphology by using low cost carbon and nitrogen sources. Firstly, a full factorial statistical design, with the factors of, two taxonomically different strains, seven types of seed culture formulations (slants) and two types of fermentation media were used to investigate the effect of these parameters on the polygalacturonase (PG) production. According to statistical analysis, factors of strain types and fermentation media and the interaction between them were found significant on the enzyme activity. Aspergillus sojae in a complex media, inoculated with a seed culture prepared from molasses resulted in maximum PG activity (0.2 U/ml). Then, a two step optimization procedure with four factors (concentrations of maltrin and corn steep liquor (CSL), agitation speed and inoculation ratio) was used to investigate the effect of these parameters on the PG activity, mycelia growth (biomass) and morphology (pellet size) of Aspergillus sojae. According to the results of response surface methodology (RSM), concentrations of maltrin, CSL and agitation speed were significant (p<0.05) on both PG synthesis and biomass formation. As a result, maximum PG activity (13.5 U/ml) was achievable at high maltrin (120 g/l), low CSL (0 g/l), high agitation speed (350 rpm) and high inoculation ratio (2x107 total spore). The diameter of pellets ranged between 0.05-0.63 cm. The second optimization step improved the PG activity by 74 % and the biomass by 40 %. Furthermore characterization of the enzyme with respect to its optimum pH and temperature and the effect of these on the stability were considered. Determination of the thermal inactivation constant with its inactivation energy and the substrate specificity constant were estimated.Master Thesis Effects of Trace Elements on the Production of Baker's Yeast(Izmir Institute of Technology, 2002) Üzelyalçın, Berna; Harsa, Hayriye ŞebnemBaker.s yeast has been used widely in traditional bakeries to produce different kinds of baked products and for many biotechnological purposes. The principal raw materials used in producing baker.s yeast are the pure yeast culture and molasses. Cane molasses and beet molasses contain 45 to 55 % sugar in the forms of sucrose, glucose, and fructose. Molasses, which is a rich carbon and mineral source, contains most of the nutrients and minerals required for baker.s yeast production. Nutrients and trace elements (magnesium, copper and zinc) that are not present in sufficient quantities in molasesses are added to the fermentation medium. In this study, the effects of Ca2+, Mg2+, Cu2+ and Zn2+ on baker.s yeast production were investigated. The method of Food Composition Laboratory (FCL), Human Nutrition Research Center, Beltsville, US, was used for the preparation of the fermentation samples prior to ICP-AES analysis. The concentrations of Ca2+, Mg2+, Cu2+, Zn2+ were determined in the raw materials entering the fermentors and the products. FCL digestion method was evaluated on precision and accuracy; linear standard curves were obtained for the studied concentration ranges of Ca2+, Mg2+, Cu2+ and Zn2+. Standard addition curves for each element in 45 brix molasses and dry yeast samples were constructed and spike recovery percentages were calculated. High spike recovery percentages were obtained for all elements in both dry yeast and molasses with the FCL method. In industrial fermentations, uptakes of Ca2+, Cu2+, Mg2+ and Zn2+ by the yeast cells were calculated as mg/kg dry yeast at 4h time intervals. The uptake rates were evaluated based on the available element concentration in the medium and the uptake of the elements. Results showed that, as the uptake of Ca2+ increased with time, uptake of Mg2+, Cu2+ and Zn2+ decreased. The yeast cells used the available elements with decreasing uptake rates except Ca2+. During the first hours of fermentation, uptake rates were high. The maximum uptake rates observed for each element in two different industrial fermentations were: 17.73-23.32 mg Cu2+/ kg dry yeast cells , 195.64-413.46 mg Zn2+ / kg dry yeast cells, 2106.46-2051.74 mg Mg2+ / kg dry yeast cells. Ion concentrations of biomass in industrial fermentations revealed that same amounts of Mg were taken by the cell from the fermentation medium under similar conditions. This was verified by the studies done in lab-scale using pure culture of S.cerevisiae, grown on different concentrations of Mg2+. It was found that uptake of Mg2+ was constant and very close to the uptake values of industrial fermentations. 806.7 mM Mg2+ was taken by 1 kg of dry yeast cells. This is in agreement with the stated Mg2+ uptake concentration range 500-1000 mM of Saccharomyces cerevisiae in literature. In the industry, magnesium is added in the form of Mg.SO4.7H2O to the fermentation medium. SO42- ions coming from the MgSO4.7H2O cause settling problems in the waste treatment system of the plant. Generally, molasses contain sufficient concentration of Mg2+ ions required for baker.s yeast production in the industrial scale. Since Mg2+ has toxic effects at high concentrations and its addition is increasing the cost of the process and causing difficulties in the waste treatment of the plant, it may be concluded that addition of Mg2+ as MgSO4.7H2O to the fermentation medium should be done after determining the Mg2+ concentration of molasses. Instead of using excessive amounts of MgSO4.7H2O, only the required quantity should be added to the fermentation medium.Master Thesis Investigation of the Effects of Dissolved Oxygen Concentration, Aeration and Agitation on the Morphology and Rheology in Submerged Fungal Fermentation(Izmir Institute of Technology, 2007) Öncü, Şelale; Ünlütürk, SevcanThe effects of pH, agitation speed, dissolved oxygen tension (DOT) and aeration, significant in common fungal fermentations, on the production of polygalacturonase (PG) enzyme and their relation to morphology and broth rheology were investigated in a batch bioreactor using Aspergillus sojae which has no available literature report on the pectinase production. All four factors were effective on the response parameters under study. An uncontrolled pH increased biomass and PG activity by 27% and 38%, respectively compared to controlled pH (pH 6). pH did not significantly affect the broth rheology but created an impact on the pellet morphology. Similarly, the maximum biomass obtained at 500 rpm and at 30h was 3.27 and 3.67 times more than at 200 and 350 rpm, respectively. The maximum enzyme productivity of 0.149 U ml-1 h-1 was obtained at 200 rpm. Non . Newtonian and pseudoplastic broth rheology was observed at 500 rpm agitation speed. Furthermore, a DOT range of 30-50% was essential for maximum biomass formation, whereas only 10% DOT was required for maximum PG synthesis. Non . Newtonian shear thickening behavior (n>1.0) was depicted at DOT levels of 10% and 30%, whereas, non-Newtonian shear thinning behavior (n<1.0) was dominant at 50% DOT. When 2.5 l/min aeration experiment was investigated detaily; it was determined that at about 21st hour, polygalacturonase production approaches its maximum (1.49 U) and pellets are smaller, high in number. At 48th hour; polygalacturonase production declines to zero, biomass reaches its maximum and pellets are big (average pellet size is 1.94±0.58 mm) and fluffy with compact centers. At the end of fermentation (96.hour), fermentation medium is close to Newtonian. The overall fermentation duration (50-70h) was considerably shorter as opposed to common fungal fermentations revealing the economic feasibility of this particular process. As a result this study not only introduced a new strain with a potential of producing a highly commercially significant enzyme but also provided certain parameters significant in the design and mathematical modelling of fungal bioprocesses.Master Thesis Determination of Whey-Based Medium Requirements and Growth Characteristics for the Production of Yoghurt Starter Cultures(Izmir Institute of Technology, 2008) Soydemir, Elçin; Harsa, Hayriye ŞebnemYoghurt is an important dairy product which is produced by thermophilic starter cultures including; Streptococcus thermophilus and Lactobacillus bulgaricus. They grow synergistically in milk by acidifying milk, produce specific aroma compounds and lactic acid which impact the quality of the yoghurt. Imported cultures are widely used in dairy industry to stabilize the technological properties which cause a change in our natural flora and flavour. Starters from traditional yoghurts have to be preserved. Under this perspective, the aim of this study was to determine the medium requirements of mixed starter cultures for production of starter bacteria which has been isolated and characterized before.Determination of medium requirements were done in a whey-based media. Since bacteriophages were one of the most important problem in dairy industry, to prevent cultures from phage adsorption, media was treated with different salts which can bind free calcium ions. Salts were examined for cell viability and phage infection effects.From these studies Na2HPO4 + KH2 PO4 with 2% of medium addition was chosen as the most effective. Mineral requirements of the mixed cultures were determined. The best combination; 10, 5, 50 ppm for Fe2+, Mn2+, Mg2+ ions with 0,939 desirability were used for initial mineral content of medium. Fermentation constants for mixed cultures were determined. Pure culture fermentations and mixed culture fermentations were performed. According to results mixed culture inoculations gave higher maximum specific growth rates; 0,9188h-1 and 0,7323h-1 for cocci and bacilli while 0,855h-1 and 0,659h-1 were obtained for cocci and bacilli from single strain fermentations.Master Thesis Bioethanol Production From Fungal Sources Using Low-Cost Agro-Industrial Waste Products(Izmir Institute of Technology, 2012) Evcan, Ezgi; Tarı, Canan; Özen, Fatma BanuIn recent years, the rapid increase in environmental problems, greenhouse gas emissions, fuel prices and the unlimited consumption of fuel stocks made people search for some alternative energy sources. Bioethanol is one of the most popular alternative sources with its many beneficial features. Considering the sugar content of fruit pomaces, which are the waste of fruit juice industry, are very convenient and cheap fermentation raw materials for production of bioethanol. The aim of this study was to create a renewable alternative for fossil fuel and to provide a viable solution to multiple environmental problems simultaneously creating a sink for waste utilization and optimize bioethanol production from apple pomace hydrolysate using Trichoderma harzianum, Aspergillus sojae and Saccharomyces cerevisiae by statistical methods. Here, screening and optimization steps were conducted in order to determine the significant factors and their optimum levels. Factors such as inoculation rate of A.sojae and T.harzianum and agitation speed were considered as factor variables, whereas the response variable was bioethanol production. According to the results of the screening process, inoculation rate of S.cerevisiae was fixed as 4% and aeration method as vented. In the optimization step, levels of the other factors were enlarged. The highest bioethanol production and yield on substrate were 8.748 g/l and 0.946, respectively. Higher concentrations of inoculation rates of T.harzianum and A.sojae (6%) and agitation speed of 200 rpm led to maximum bioethanol production. Furthermore, the results pointed out that using cocultures because of its synergistic interactions is an effective way for production of bioethanol.