Food Engineering / Gıda Mühendisliği
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Research Project Aspergillus sojae tarafından poligalakturonaz (PG) enzimin üretimi ve pürifikasyonu: Morfoloji ve sıvı reolojisinin toplam sistem verimliliği üzerine olan etkilerinin endüstriyel uygulamalara yakın ölçeklerde deneysel ve teorik yaklaşımla incelenmesi(2011) Tarı, Canan; Ünlütürk, Sevcan[No Abstract Available]Article Effect of Substrate Concentration and Scale Up on the Polygalacturonase Production(Gıda Teknolojisi Derneği, 2019) Göğüş, Nihan; Tarı, CananPectinases have been used for many industrial applications since long time ago. The largest industrial application of these enzymes is in fruit juice and wine production for the extraction, filtration and clarification and for the maceration of fruits and vegetables. They work by enzymatic breaking down of the cell wall.In this study it was aimed to use the previously optimized shake flask media formulation in batch mode 1 L scale serial bioreactor system and 5 L scale in order to investigate the effects of substrate concentration and scale on PG activity and biomass production. In conclusion it was observed that average PG activity (101.29 U/ml) obtained in 5L scale bioreactor experiments was higher than the maximum PG activity (88.55 U/ml) at 40 g/L orange peel (OP) concentration obtained in the 1 L scale substrate concentration experiment. Furthermore,PG activity increased with an increase in substrate concentration except for 60 g/L orange peel concentration.Article Citation - WoS: 69Citation - Scopus: 89Pectinase Enzyme-Complex Production by Aspergillus Spp. in Solid-State Fermentation: a Comparative Study(Elsevier, 2012) Heerd, Doreen; Yeğin, Sırma; Tarı, Canan; Fernandez Lahore, MarceloA comparative evaluation of three Aspergillus species according to their pectinase production in solid-state fermentation was performed. Solid-state fermentation offers several potential advantages for enzyme production by fungal strains. Utilization of agricultural by-products as low-cost substrates for microbial enzyme production resulted in an economical and promising process. The pectinolytic enzyme activities of two Aspergillus sojae strains were compared to a known producer, Aspergillus niger IMI 91881, and to A. sojae ATCC 20235, which was re-classified as Aspergillus oryzae. Evaluation of polymethylgalacturonase and polygalacturonase activity was performed as well as exo- vs. endo-enzyme activity in the crude pectinase enzyme-complex of the mentioned strains. Furthermore, a plate diffusion assay was applied to determine the presence and action of proteases in the crude extracts. A. sojae ATCC 20235 with highest polymethylgalacturonase activity and highest polygalacturonase activity both exo- and endo-enzyme activity, is a promising candidate for industrial pectinase production, a group of enzymes with high commercial value, in solid-state fermentation processes. Beside the enzymatic assays a protein profile of each strain is given by SDS-PAGE electrophoresis and in addition species-specific zymograms for pectinolytic enzymes were observed, revealing the differences in protein pattern of the A. sojae strains to the re-classified A. oryzae. (C) 2011 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.Article Aspergillus Sojae Tarafından Üretilen Poligalakturonazın Kısmi Saflaştırılması için Kromatografik Bir Yaklaşım(Sidas Medya Ajans Tanıtım Danışmanlık Ltd. Şti., 2017) Şen, İlknur; Mata-Gomez, Marco A.; Palomares, Marco Rito; Tarı, Canan; Dinç, MelikeBu çalışmanın amacı, A. sojae mutantından poligalakturonaz üretilmesi ve ham ekstraktın kromatografik yöntemlerle kısmi saflaştırılmasıdır. Peptitlerin konfirmasyonu için ilk basamak olarak, jel içinde sindirilmiş sodyum-dodesil-sülfatpoliakrilamid-jel-elektroforezi (SDS-PAGE) jellerinde matriks-yardımlı lazer desorpsiyon/iyonlaştırmalı-uçuş zamanlıkütle spektrometresi (Maldi-TOF MS) analizi yapılmıştır. Poligalakturonaz üretimi için, katı-faz ve derin fermentasyonlarda üç farklı karbon kaynağı kullanılmıştır. Ham ekstrakt ilk olarak iyon değişim kromatografisi (IEXC) ile saflaştırılmıştır ve ardından bunu boyut eleme kromatografisi izlemiştir. Derin [acı portakal kabuğu, şeker pancarı melası ve (NH4)2SO4] ve katı-faz (buğday kepeği, şeker pancarı ve HCl) fermentasyonlarından elde edilen ham ekstraktlar yüksek seviyede poligalakturonaz enzim aktivitesi (sırasıyla 95.22 and 50.27 U/mL) göstermiştir. IEXC toplanmış fraksiyonunun (180, 200 ve 220 mM tuz fraksiyonları) boyut elemesi, en yüksek verimi (%36) ve saflaştırma katını (2.00) göstermiştir. SDS-PAGE'den elde edilen olası poligalakturonaz bantları jel içinde sindirilmiş ve peptit konfirmasyonu için Maldi-TOF-MS ile analiz edilmiştir.Article Citation - WoS: 18Citation - Scopus: 35Microparticle-Enhanced Polygalacturonase Production by Wild Type Aspergillus Sojae(Springer Verlag, 2017) Karahalil, Ercan; Demirel, Fadime; Evcan, Ezgi; Germeç, Mustafa; Tarı, Canan; Turhan, İrfanPolygalacturonases (PGs), an important industrial enzyme group classified under depolymerases, catalyze the hydrolytic cleavage of the polygalacturonic acid chain through the introduction of water across the oxygen bridge. In order to produce and increase the concentration of this enzyme group in fermentation processes, a new approach called microparticle cultivation, a promising and remarkable method, has been used. The aim of this study was to increase the PG activity of Aspergillus sojae using aluminum oxide (Al2O3) as microparticles in shake flask fermentation medium. Results indicated that the highest PG activity of 34.55 ± 0.5 U/ml was achieved with the addition of 20 g/L of Al2O3 while the lowest activity of 15.20 ± 0.2 U/mL was obtained in the presence of 0.1 g/L of Al2O3. In fermentation without microparticles as control, the activity was 15.64 ± 3.3 U/mL. Results showed that the maximum PG activity was 2.2-fold higher than control. Additionally, smaller pellets formed with the addition of Al2O3 where the lowest pellet diameter was 955.1 µm when 10 g/L of the microparticle was used. Also, it was noticed that biomass concentration gradually increased with increasing microparticle concentration in the fermentation media. Consequently, the PG activity was significantly increased in microparticle-enhanced shake flask fermentation. In fact, these promising preliminary data can be of significance to improve the enzyme activity in large-scale bioreactors.Article Citation - WoS: 1Citation - Scopus: 2Partial Purification of a Polygalacturonase From a New Aspergillus Sojae Mutant and Its Application in Grape Mash Maceration(John Wiley and Sons Inc., 2017) Yıldız, Semanur; Mata-Gomez, Marco A.; Tarı, Canan; Rito-Palomares, MarcoThe use of polygalacturonase (PG) preparations in winemaking promotes the release of phenolic compounds. A PG from a new source, Aspergillus sojae mutant, was semi-purified and tested for grape mash maceration. Crude extract (CE), a commercial pectinase, and two high PG activity semi-purified preparations, FI and FII , were applied for maceration at PG activity of 3.5 U g−1 of grape for 46 h. Enzyme-assisted maceration significantly (P < 0.05) increased the total phenolic content from 255.8 to 916.3 ± 5.2, 5732.9 ± 9.9, 563.4 ± 6.7 and 620.6 ± 18.4 mg L−1 for CE, commercial pectinase, FI and FII, respectively. The content of individual phenolics such as gallic, protocatechuic, chlorogenic and p-coumaric acids was improved. Principal component and hierarchical clustering analyses suggested that CE has a better performance upon the release of phenols. Semi-purified preparations acted similar to commercial pectinase. These findings open an opportunity for the potential use of PG from the mutant strain as an alternative macerating enzyme.Article Citation - WoS: 13Citation - Scopus: 18Effect of Physicochemical Parameters on the Polygalacturonase of an Aspergillus Sojae Mutant Using Wheat Bran, an Agro-Industrial Waste, Via Solid-State Fermentation(John Wiley and Sons Inc., 2016) Demir, Hande; Tarı, CananBACKGROUND: Polygalacturonases (PGs) are valuable enzymes of the food industry; therefore it is of great importance to discover new and GRAS PG-producing microbial strains. In this study, PG enzyme produced from a high PG activity producer mutant Aspergillus sojae using wheat bran at the flask scale under pre-optimized conditions of solid-state fermentation (SSF) was biochemically characterized. RESULTS: The crude PG enzyme showed optimum activity in the pH range 4.0–5.0 and was stable in the pH range 3.0–7.0. The optimum temperature for the PG was 40 °C and it retained 99% of its activity at 50 °C. The mutant A. sojae PG could preserve more than 50% of its stability between 25 and 50 °C, both for 30 and 60 min, and was found to be stable in the presence of most of the tested compounds and metal ions. The inactivation energy (Ed) was determined as 125.3 kJ mol−1. The enthalpy (ΔH*), free energy (ΔG*) and entropy (ΔS*) of inactivation were found to be stable with increasing temperature. CONCLUSION: The mutant A. sojae PG could be suitable for the clarification (depectinization) of orange and grape juices and wine. © 2015 Society of Chemical Industry.Article Citation - WoS: 12Citation - Scopus: 16Utilization of Orange Peel, a Food Industrial Waste, in the Production of Exo-Polygalacturonase by Pellet Forming Aspergillus Sojae(Springer Verlag, 2015) Büyükkileci, Ali Oğuz; Lahore, Marcelo Fernandez; Tarı, CananThe production of exo-polygalacturonase (exo-PG) from orange peel (OP), a food industrial waste, using Aspergillus sojae was studied in submerged culture. A simple, low-cost, industrially significant medium formulation, composed of only OP and (NH4)2SO4 (AS) was developed. At an inoculum size of 2.8 × 103 spores/mL, growth was in the form of pellets, which provided better mixing of the culture broth and higher exo-PG activity. These pellets were successfully used as an inoculum for bioreactors and 173.0 U/mL exo-PG was produced. Fed-batch cultivation further enhanced the exo-PG activity to 244.0 U/mL in 127.5 h. The final morphology in the form of pellets is significant to industrial fermentation easing the subsequent downstream processing. Furthermore, the low pH trend obtained during this fermentation serves an advantage to fungal fermentations prone to contamination problems. As a result, an economical exo-PG production process was defined utilizing a food industrial by-product and producing high amount of enzyme.Article Citation - WoS: 52Citation - Scopus: 72Production of Bioethanol From Apple Pomace by Using Cocultures: Conversion of Agro-Industrial Waste To Value Added Product(Elsevier Ltd., 2015) Evcan, Ezgi; Tarı, CananDirect fermentation of cellulosic biomass to bioethanol has been very promising and hence attracted attention in recent years. In this study, bioethanol production from apple pomace hydrolysate (agro-industrial waste product) was investigated by coculturing Trichoderma harzianum, Aspergillus sojae and Saccharomyces cerevisiae using statistical approaches. Screening and optimization experiments were conducted in order to determine the significant factors and their optimum levels for maximum bioethanol production. Inoculation rates, aeration and agitation speed were considered as factor variables and bioethanol production as response variable. Highest bioethanol (EtOH) concentration and ethanol yield on total reducing sugar content (YP/S) were 8.748 g/L and 0.945 g/g, respectively. Optimum conditions were 6% (w/v) inoculation rates of T.harzianum and A.sojae, and 4% (v/v) inoculation rate of S.cerevisiae with vented aeration method and agitation speed of 200 rpm. To best of our knowledge to date, no reports are available in literature regarding the coculturing of T.harzianum, A.sojae and S.cerevisiae for bioethanol production. Therefore, this study will serve as a base line of initial studies in this field. The method can create a renewable alternative feedstock for fossil fuel production and suggest a feasible solution to multiple environmental problems simultaneously creating a sink for waste utilization.Article Citation - WoS: 28Citation - Scopus: 38Bioconversion of Wheat Bran for Polygalacturonase Production by Aspergillus Sojae in Tray Type Solid-State Fermentation(Elsevier Ltd., 2016) Demir, Hande; Tarı, CananWheat bran was tested as the solid substrate for the tray-type solid-state fermentation (SSF) production of polygalacturonase (PG) enzyme by A. sojae mutant strain - a high-PG activity producer. PG production of A. sojae was found to reduce as the thickness of the substrate increase from 8 mm to 14 mm at 90% relative humidity. An interaction between the thickness of the bed and relative humidity of the environment was determined with the help of experimental design and statistical analysis tools. As a result, the PG activity could be enhanced by 31% as the process conditions optimized. Additionally, 11 mm thickness and 70% relative humidity were selected as the PG production favoring conditions with the maximum PG activity of 298 U/g substrate in tray type of SSF without the addition of any nutritive or inducing supplements into wheat bran. The kinetic study conducted in the trays revealed the presence of reduction in the water activity on the 4th day of the SSF process under stated conditions. The productivity of the process conducted under optimized conditions was 3.41 U/g substrate-1 h-1 for the 4th day of the SSF. © 2015 Elsevier Ltd.
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