Food Engineering / Gıda Mühendisliği
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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 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: 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: 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.Article Citation - WoS: 14Citation - Scopus: 12Enhanced Production of Exo-Polygalacturonase From Agro-Based Products by Aspergillus Sojae(North Carolina University, 2011) Büyükkileci, Ali Oğuz; Tarı, Canan; Fernandez-Lahore, MarcelloAspergillus sojae has been previously shown to produce exo-polygalacturonase (exo-PG) in synthetic media, where the potential of the organism to utilize agricultural substrates was not considered so far. In this study, the utilization of agro-based products was taken into account in the enhanced production of exo-PG using an A. sojae mutant by applying statistical design methods. Complex sources (orange peel, wheat bran, and corn meal), simple sugar sources (glucose, maltrin, and sugar beet syrup), and two phosphate salts were screened using D-optimal design method. Orange peel yielded the highest exo-PG activity with all simple sugars and phosphate sources. According to the results of response surface methodology (RSM), the optimum concentrations of orange peel, sugar beet syrup, and (NH 4) 2SO 4 were found to be 10, 60, and 8 g L -1, respectively. The exo-PG activity under these conditions was 145.4 U m L -1 in shake flask cultures. In bioreactor studies enzyme production was induced at low pH values; thus highest production was obtained under uncontrolled pH conditions, in which the pH dropped to 2.0 in 72 h. As a result high exo-PG could be produced by an A. sojae mutant using a cost-effective medium containing agro-industrial substrates. Another important advantageous outcome was the low optimal pH, which is especially desired in industrial fermentations prone to contamination problems. In fact this highlights the easy adaptation of this fermentation to industrial scales.Article Citation - WoS: 50Citation - Scopus: 59Characterization of Three-Phase Partitioned Exo-Polygalacturonase From Aspergillus Sojae With Unique Properties(Elsevier Ltd., 2008) Doğan, Nergiz; Tarı, CananExo-polygalacturonase enzyme produced by Aspergillus sojae ATCC 20235 was purified using three-phase partitioning (TPP), an emerging bio-separation technique where a single step as compared to the classical multi-step purification was used. Using this technique, crude enzyme solution (pH 6.6) saturated to 30% (w/v) with ammonium sulphate and with a crude extract to tert-butanol ratio of 1:1 (v/v) at 25 °C resulted in 25.5% recovery of exo-polygalacturonase with a 6.7-fold purification. The purified enzyme was characterized with respect to its activity and stability at various pH and temperature ranges. Optimum pH and temperature for maximum activity were determined as pH 4 and 55 °C. The enzyme was stable at both acidic and alkaline pH for 2 h at 30 °C. The thermal stability study showed that the purified enzyme had an inactivation energy of 68.41 kcal/mol and a half-life (t1/2) value of 3.6 h at 75 °C presenting a large thermal stability. The kinetic constants Km and Vmax using polygalacturonic acid as substrate were 0.75 g l-1 and 1.14 μmol min-1, respectively. SDS-PAGE profiling revealed that the purified exo-polygalacturonase had two bands with the molecular weights of 36 and 53 kDa. The enzyme was completely inhibited in the presence of Mn2+ and SDS and induced significantly by EDTA, glycerol and β-mercaptoethanol.Article Citation - WoS: 52Citation - Scopus: 62Solid-State Production of Polygalacturonase by Aspergillus Sojae Atcc 20235(Elsevier Ltd., 2007) Üstok, Fatma Işık; Tarı, Canan; Göğüş, NihanThe effect of solid substrates, inoculum and incubation time were studied using response surface methodology (RSM) for the production of polygalacturonase enzyme and spores in solid-state fermentation using Aspergillus sojae ATCC 20235. Two-stage optimization procedure was applied using D-optimal and face-centered central composite design (CCD). Crushed maize was chosen as the solid substrate, for maximum polygalacturonase enzyme activity based on D-optimal design. Inoculum and incubation time were determined to have significant effect on enzyme activity and total spore (p < 0.01) based on the results of CCD. A second order polynomial regression model was fitted and was found adequate for individual responses. All two models provided an adequate R2 of 0.9963 (polygalacturonase) and 0.9806 (spores) (p < 0.001). The individual optimum values of inoculum and incubation time for maximum production of the two responses were 2 × 107 total spores and 5-6 days. The predicted enzyme activity (30.55 U/g solid) and spore count (2.23 × 107 spore/ml) were very close to the actual values obtained experimentally (29.093 U/g solid and 2.31 × 107 spore/ml, respectively). The overall optimum region considering the two responses together, overlayed with the individual optima. Solid-state fermentation provided 48% more polygalacturonase activity compared to submerged fermentation under individually optimized conditions.Article Citation - WoS: 61Citation - Scopus: 75Optimization of Biomass, Pellet Size and Polygalacturonase Production by Aspergillus Sojae Atcc 20235 Using Response Surface Methodology(Elsevier Ltd., 2007) Tarı, Canan; Göğüş, Nihan; Tokatlı, FigenA two-step optimization procedure using central composite design with four factors (concentrations of maltrin and corn steep liquor (CSL), agitation speed and inoculation ratio) was used in order to investigate the effect of these parameters on the polygalacturonase (PG) enzyme activity, mycelia growth (biomass) and morphology (pellet size) of Aspergillus sojae ATCC 20235. According to the results of response surface methodology (RSM), initial concentrations of maltrin and CSL and agitation speed were significant (p < 0.05) on both PG enzyme production and biomass formation. As a result of this optimization, maximum PG activity (13.5 U/ml) was achievable at high maltrin (120 g/l), at low CSL (0 g/l), high agitation speed (350 rpm) and high inoculation ratio (2 × 107 total spore). Similarly, maximum biomass (26 g/l) could be obtained under the same conditions with only the difference for higher level of CSL requirement. The diameter of pellets in all optimization experiments ranged between 0.05 and 0.76 cm. The second optimization step improved the PG activity by 74% and the biomass by 40%.Article Citation - WoS: 33Citation - Scopus: 37Relationship Between Morphology, Rheology and Polygalacturonase Production by Aspergillus Sojae Atcc 20235 in Submerged Cultures(Elsevier Ltd., 2006) Göğüş, Nihan; Tarı, Canan; Öncü, Şelale; Ünlütürk, Sevcan; Tokatlı, FigenA 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 morphology and polygalacturonase production. The rheology of the final fermentation medium was analyzed and appropriate mathematical model was applied to calculate suspension viscosity. It was found that most fermentation broths showed non-Newtonian flow behavior. According to statistical analysis, factors of strain types and fermentation media and the interaction between them were found significant on the enzyme activity. The effect of seed culture formulations (slants) were found insignificant at the significance level of 1%. Interaction of slants with strain types and fermentation media were also found insignificant. Considering the morphology of the final culture, Aspergillus sojae with the desired pellet morphology in a complex media, inoculated with a seed culture prepared from molasses resulted in maximum polygalacturonase enzyme activity (0.2 U/ml) and lowest suspension viscosity with a broth rheology close to Newtonian flow behavior.