Use of Cassava Peel as Carbon Source for Production of Amylolytic Enzymes by Aspergillus niveus

2009 ◽  
Vol 5 (5) ◽  
Author(s):  
Tony Marcio Silva ◽  
Ricardo Fernandes Alarcon ◽  
Andre Ricardo de Lima Damasio ◽  
Michele Michelin ◽  
Alexandre Maller ◽  
...  
Keyword(s):  
Carbon Source ◽  
Crude Extract ◽  
Soluble Starch ◽  
Culture Conditions ◽  
Amylolytic Enzymes ◽  
Agricultural Residue ◽  
Ph Values ◽  
Aspergillus Niveus ◽  
Static Conditions ◽  
Cassava Peel

Aspergillus niveus produced high levels of ?-amylase and glucoamylase in submerged fermentation using the agricultural residue cassava peel as a carbon source. In static conditions, the amylase production was substantially greater than in the agitated condition. The optimized culture conditions were initially at pH 5.0, 35°C during 48 hours. Amylolytic activity was still improved (50%) with a mixture of cassava peel and soluble starch in the proportion 1:1 (w/w). The crude extract exhibited temperature and pH optima approximately 70°C and 4.5, respectively. Amylase activity was stable for 1 h at 60°C, and at pH values between 3.0 and 7.0. The enzyme hydrolysed preferentially maltose, starch, penetrose, amylose, isomaltose, maltotriose, glycogen and amylopectin, and not hydrolysed cyclodextrin (? and ß), trehalose and sucrose. In the first hour of reaction on soluble starch, the hydrolysis products were glucose and maltose, but after two hours of hydrolysis, glucose was the unique product formed, confirming the presence in the crude extract of an ?-amylase and a glucoamylase.

Statistical Analysis of Growth and Amylase Production by Aspergillus flavus grown on Different Carbon Sources

2013 ◽  
Vol 2 (1) ◽  
Author(s):  
M O Oyewale
Keyword(s):  
Carbon Source ◽  
Aspergillus Flavus ◽  
Amylase Activity ◽  
Carbon Sources ◽  
Dry Weight ◽  
Soluble Starch ◽  
Optimum Growth ◽  
Amylase Production ◽  
Level Of Significance ◽  
Cassava Peel

The mycelial dry weight and dinitrosalicylic acid (D.N.S.A.) method was used to determine growth and amylase production by Aspergillus flavus grown on different carbon sources. Growth of the fungus was determined at 24 h intervals over a period of six days by the dry mycelial weight methods, while the amylase activity in the culture filtrates of A. flavus was determined by the D.N.S.A method. A total of 45 samples were prepared to determine growth and amylase activity of Aspergillus flavus grown on different carbon sources. The concentration of the various carbon sources ranges between 0.4 to 2% W/V. Duncan’s multiple range test was used to determine the level of significance of the different carbon sources for effective growth and amylase production by Aspergillus flavus. Aspergillus flavus demonstrated the capability to produce significant growth and amylase activities in the medium containing soluble starch, sorghum and cassava peel as sole carbon source. The amount of mycelial dry weight produced from soluble starch, sorghum and cassava peel is significantly higher than those produced from other carbon sources. The data revealed that there is a correlation between growth and amylase production by Aspergillus flavus. The available data from this study showed that soluble starch is the best carbon source for optimum growth and amylase production by A flavus while sorghum and cassava peel are close substitute for optimum growth and amylase production by Aspergillus flavus. Keywords: Growth, amylase activity and Aspergillus flavus

scholarly journals Aeration and Stirring in Yarrowia lipolytica Lipase Biosynthesis during Batch Cultures with Waste Fish Oil as a Carbon Source

Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 88
Author(s):  
Paulina Snopek ◽  
Dorota Nowak ◽  
Bartłomiej Zieniuk ◽  
Agata Fabiszewska
Keyword(s):  
Carbon Source ◽  
Fish Oil ◽  
Yarrowia Lipolytica ◽  
Lipolytic Activity ◽  
Culture Conditions ◽  
Agitation Speed ◽  
Oxygen Deficit ◽  
Shear Stresses ◽  
Process Selection ◽  
Batch Cultures

Yarrowia lipolytica is one of the most studied non-conventional forms of yeast, exhibiting a high secretory capacity and producing many industrially important and valuable metabolites. The yeast conceals a great biotechnological potential to synthesize organic acids, sweeteners, microbial oil, or fragrances. The vast majority of bioprocesses are carried out in bioreactors, where suitable culture conditions are provided. In the current study, the effect of agitation speed (200–600 rpm) and air flow rate (0.0375–2.0 dm3/(dm3 × min)) on the biomass yield and lipase activity of Y. lipolytica KKP 379 is analyzed in a growth medium containing waste fish oil. The increase of aeration intensity limited the period of oxygen deficit in the medium. Simultaneously, an increase in lipolytic activity was observed from 2.09 U/cm3 to 14.21 U/cm3; however, an excessive agitation speed likely caused oxidative or shear stresses, and a reduction in lipolytic activity was observed. Moreover, it is confirmed that the synthesis of lipases is related to oxygen consumption, pH, and the yeast growth phase, and appropriate process selection may provide two advantages, namely, the maximum use of the waste carbon source and the production of lipolytic enzymes that are valuable in many industries.

scholarly journals Mimicking of Blood Flow Results in a Distinct Functional Phenotype in Human Non-Adherent Classical Monocytes

Biology ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 748
Author(s):  
Elisa Wirthgen ◽  
Melanie Hornschuh ◽  
Ida Maria Wrobel ◽  
Christian Manteuffel ◽  
Jan Däbritz
Keyword(s):  
Blood Flow ◽  
Shear Flow ◽  
Ex Vivo ◽  
Culture Conditions ◽  
Inflammatory Signaling ◽  
Gm Csf ◽  
Beneficial Effects ◽  
Migratory Capacity ◽  
Ex Vivo Culture ◽  
Static Conditions

Ex vivo culture conditions during the manufacturing process impact the therapeutic effect of cell-based products. Mimicking blood flow during ex vivo culture of monocytes has beneficial effects by preserving their migratory ability. However, the effects of shear flow on the inflammatory response have not been studied so far. Hence, the present study investigates the effects of shear flow on both blood-derived naïve and activated monocytes. The activation of monocytes was experimentally induced by granulocyte-macrophage colony-stimulating factor (GM-CSF), which acts as a pro-survival and growth factor on monocytes with a potential role in inflammation. Monocytes were cultured under dynamic (=shear flow) or static conditions while preventing monocytes' adherence by using cell-repellent surfaces to avoid adhesion-induced differentiation. After cultivation (40 h), cell size, viability, and cytokine secretion were evaluated, and the cells were further applied to functional tests on their migratory capacity, adherence, and metabolic activity. Our results demonstrate that the application of shear flow resulted in a decreased pro-inflammatory signaling concurrent with increased secretion of the anti-inflammatory cytokine IL-10 and increased migratory capacity. These features may improve the efficacy of monocyte-based therapeutic products as both the unwanted inflammatory signaling in blood circulation and the loss of migratory ability will be prevented.

scholarly journals Synthesis of the Microbial Polysaccharide Gellan from Dairy and Plant-Based Processing Coproducts

2021 ◽  
Vol 2 (2) ◽  
pp. 234-244
Author(s):  
Thomas P. West
Keyword(s):  
Corn Steep Liquor ◽  
Nitrogen Sources ◽  
Soluble Starch ◽  
Culture Conditions ◽  
Water Soluble ◽  
Production Costs ◽  
Microbial Polysaccharide ◽  
Steep Liquor ◽  
The Cost ◽  
Condensed Distillers Solubles

This review examines the production of the microbial polysaccharide gellan, synthesized by Sphingomonas elodea, on dairy and plant-based processing coproducts. Gellan is a water-soluble gum that structurally exists as a tetrasaccharide comprised of 20% glucuronic acid, 60% glucose and 20% rhamnose, for which various food, non-food and biomedical applications have been reported. A number of carbon and nitrogen sources have been tested to determine whether they can support bacterial gellan production, with several studies attempting to optimize gellan production by varying the culture conditions. The genetics of the biosynthesis of gellan has been explored in a number of investigations and specific genes have been identified that encode the enzymes responsible for the synthesis of this polysaccharide. Genetic mutants exhibiting overproduction of gellan have also been identified and characterized. Several dairy and plant-based processing coproducts have been screened to learn whether they can support the production of gellan in an attempt to lower the cost of synthesizing the microbial polysaccharide. Of the processing coproducts explored, soluble starch as a carbon source supported the highest gellan production by S. elodea grown at 30 °C. The corn processing coproducts corn steep liquor or condensed distillers solubles appear to be effective nitrogen sources for gellan production. It was concluded that further research on producing gellan using a combination of processing coproducts could be an effective solution in lowering its overall production costs.

scholarly journals Production and Optimization of Xylanase and α-Amylase from Non-Saccharomyces Yeasts (Pichia membranifaciens)

2021 ◽  
pp. 452-461
Author(s):  
Hala A. Salah ◽  
Hanan A. Temerk ◽  
Nivin A. Salah ◽  
Saeed Rafa Zara Alshehri ◽  
Jazi A. Al-Harbi ◽  
...  
Keyword(s):  
Culture Medium ◽  
Potato Starch ◽  
Pcr Amplification ◽  
Soluble Starch ◽  
Relative Activity ◽  
Rrna Gene ◽  
Pichia Membranifaciens ◽  
Ph Values ◽  
26S Rrna ◽  
Saccharomyces Yeasts

The xylanolytic and amylolytic yeasts were qualitatively determined by Cong red xylan agar and soluble starch agar plates, respectively. The most xylanase and α-amylase inducible strain (AUN-02) was selected and identified using PCR amplification of 26S rRNA gene and sequence analysis. The comparison of the alignment results and phylogenetic analysis of the sequences of the isolated yeast to published rRNA gene sequences in GenBank, confirmed the identification of the isolate as Pichia membranifaciens. Xylanase and α-amylase production by isolated P. membranifaciens were investigated at different pH values (4-8), temperature degrees (20-45°C), incubation time (1-7 days) and various substrates.A higher production of xylanase (38.8 U/mL) and a-amylase (28.7 U/mL) was obtained after 4 days of fermentation of P. membranifaciens. Higher activity of xylanase (36.83 U/mL) and a-amylase (27.7 U/mL) was obtained in the fermentation of P. membranifaciens in a culture medium adjusted to pH 7.0. The optimum temperature showed maximum xylanase and a-amylase activity (42.6 and 32.5 units/mL, respectively) was estimated at 35 °C. The xylanase and a-amylase activities of P. membranifaciens were estimated and compared for the different substrates tested. The strain revealed 100% relative activity of xylanase and a-amylase on beechwood and potato starch, respectively. The affinity of enzymes towards substrate was estimated using Km values. The Km values of xylanase and α-amylase increased in the order of pH’s 7.0, 6.0 and 4.5 (0.85, 1.6 and 3.4 mg xylan/mL and 0.22, 0.43 and 2.8 mg starch/mL, respectively). the yeast P. membranifaciensis is suitable for produce neutral xylanase and α-amylase enzymes. So, it could be used as a promising strain for production of these enzymes in industrial field.

scholarly journals Effect of culture conditions on nitrogen-fixing activity of bacteria isolated from cassava cultivated soils of Vietnam

2021 ◽  
Vol 43 (3) ◽  
pp. 27-35
Author(s):  
Pham Viet Cuong ◽  
Nguyen Phuong Hoa
Keyword(s):  
Bacillus Subtilis ◽  
Carbon Source ◽  
16S Rrna ◽  
Culture Conditions ◽  
Rrna Gene ◽  
Atmospheric Nitrogen ◽  
Bacillus Sp ◽  
Nitrogen Fixing ◽  
The 16S Rrna Gene ◽  
Cultivated Soils

The bacteria capable of fixing atmospheric nitrogen were isolated from cassava cultivated soils of Vietnam. The potential isolates were identified by analyzing the 16S rRNA gene and by morphological, biochemical, cultural characteristics. The selected isolates were assigned to the species Bacillus sp. DQT2 M17, Bacillus subtilis DTAN6 M17, and Bacillus megaterium DSHB I8. The effect of culture conditions on the nitrogen-fixing activity of three selected isolates were studied and the obtained results showed that the highest amount of accumulated ammonia was detected after 6 days of incubation at 35 oC, pH 7.0 with sucrose as a carbon source. The selected strains could be exploited as inoculants for microbial fertilizer production.

scholarly journals DETERMINATION OF OPTIMUM CULTURE CONDITIONS FOR MYCELIAL GROWTH OF Macrolepiota procera MUSHROOM

2020 ◽  
Vol 19 (1) ◽  
pp. 11-20
Author(s):  
Aysun Pekşen ◽  
Beyhan Kibar
Keyword(s):  
Yeast Extract ◽  
Mycelial Growth ◽  
Nitrogen Sources ◽  
Culture Conditions ◽  
Malt Extract ◽  
The Public ◽  
Carbon And Nitrogen ◽  
Ph Values ◽  
Incubation Temperatures ◽  
Macrolepiota Procera

Macrolepiota procera, commonly called the Parasol Mushroom, is a delicious mushroom collected from the nature and commonly consumed by the public in many regions of Turkey. This study was conducted to determine the optimum culture conditions (pH, temperature, carbon and nitrogen sources) for mycelial growth of M. procera. Three pH values (pH 5.0, 5.5 and 6.0), four incubation temperatures (15, 20, 25 and 30°C), seven carbon (C) sources (dextrose, glucose, lactose, maltose, mannitol, sucrose and xylose) and six nitrogen (N) sources ((NH4)2HPO4, NH4NO3 and Ca(NO3)2, malt extract, peptone and yeast extract) were investigated. In the second step of the study, the effect of seven pH values (4.0, 4.5, 5.0, 5.5, 6.0, 6.5 and 7.0) on the mycelial colony diameter was examined at 20 and 25°C since these temperatures gave the best mycelial growth in the previously conducted temperature experiment. The best mycelial growth was determined at pH 6.0. The optimum temperature for mycelial growth of M. procera was found as 25°C. The use of glucose as carbon source and yeast extract and peptone as nitrogen source in the culture medium gave the best results for mycelial growth. Determining of optimum culture conditions for mycelial growth of M. procera will provide important contributions to the fortcoming studies on it’s commercially cultivation in Turkey.

scholarly journals Gravity-Based Flow Efficient Perfusion Culture System for Spheroids Mimicking Liver Inflammation

Biomedicines ◽  
2021 ◽  
Vol 9 (10) ◽  
pp. 1369
Author(s):  
Young-Su Kim ◽  
Arun Asif ◽  
Abdul Rahim Chethikkattuveli Salih ◽  
Jae-Wook Lee ◽  
Ki-Nam Hyun ◽  
...  
Keyword(s):  
Culture System ◽  
Disease Modeling ◽  
Perfusion Culture ◽  
Drug Analysis ◽  
Culture Conditions ◽  
Current Approach ◽  
Spheroid Culture ◽  
Organ Specific ◽  
Static Conditions

The spheroid culture system provides an efficient method to emulate organ-specific pathophysiology, overcoming the traditional two-dimensional (2D) cell culture limitations. The intervention of microfluidics in the spheroid culture platform has the potential to enhance the capacity of in vitro microphysiological tissues for disease modeling. Conventionally, spheroid culture is carried out in static conditions, making the media nutrient-deficient around the spheroid periphery. The current approach tries to enhance the capacity of the spheroid culture platform by integrating the perfusion channel for dynamic culture conditions. A pro-inflammatory hepatic model was emulated using a coculture of HepG2 cell line, fibroblasts, and endothelial cells for validating the spheroid culture plate with a perfusable channel across the spheroid well. Enhanced proliferation and metabolic capacity of the microphysiological model were observed and further validated by metabolic assays. A comparative analysis of static and dynamic conditions validated the advantage of spheroid culture with dynamic media flow. Hepatic spheroids were found to have improved proliferation in dynamic flow conditions as compared to the static culture platform. The perfusable culture system for spheroids is more physiologically relevant as compared to the static spheroid culture system for disease and drug analysis.

scholarly journals Engineering large cartilage tissues using dynamic bioreactor culture at defined oxygen conditions

2018 ◽  
Vol 9 ◽  
pp. 204173141775371 ◽  
Author(s):  
Andrew C Daly ◽  
Binulal N Sathy ◽  
Daniel J Kelly
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Culture Conditions ◽  
Cartilage Tissue ◽  
Bioreactor Culture ◽  
Dynamic Culture ◽  
Oxygen Conditions ◽  
Static Conditions

Mesenchymal stem cells maintained in appropriate culture conditions are capable of producing robust cartilage tissue. However, gradients in nutrient availability that arise during three-dimensional culture can result in the development of spatially inhomogeneous cartilage tissues with core regions devoid of matrix. Previous attempts at developing dynamic culture systems to overcome these limitations have reported suppression of mesenchymal stem cell chondrogenesis compared to static conditions. We hypothesize that by modulating oxygen availability during bioreactor culture, it is possible to engineer cartilage tissues of scale. The objective of this study was to determine whether dynamic bioreactor culture, at defined oxygen conditions, could facilitate the development of large, spatially homogeneous cartilage tissues using mesenchymal stem cell laden hydrogels. A dynamic culture regime was directly compared to static conditions for its capacity to support chondrogenesis of mesenchymal stem cells in both small and large alginate hydrogels. The influence of external oxygen tension on the response to the dynamic culture conditions was explored by performing the experiment at 20% O2 and 3% O2. At 20% O2, dynamic culture significantly suppressed chondrogenesis in engineered tissues of all sizes. In contrast, at 3% O2 dynamic culture significantly enhanced the distribution and amount of cartilage matrix components (sulphated glycosaminoglycan and collagen II) in larger constructs compared to static conditions. Taken together, these results demonstrate that dynamic culture regimes that provide adequate nutrient availability and a low oxygen environment can be employed to engineer large homogeneous cartilage tissues. Such culture systems could facilitate the scaling up of cartilage tissue engineering strategies towards clinically relevant dimensions.

Abstract 139: Using Cyclic Strain to Improve Cardiac Progenitor Cell Cooperation

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Kristin M French ◽  
Marcos J Fierro ◽  
Todd D Johnson ◽  
Karen L Christman ◽  
Michael E Davis
Keyword(s):  
Connexin 43 ◽  
Matrix Protein ◽  
Mechanical Strain ◽  
Cyclic Strain ◽  
Culture Conditions ◽  
Cardiac Cells ◽  
Rhodamine Phalloidin ◽  
Cell Therapies ◽  
Static Conditions

Introduction: Cell therapies have grown in popularity for myocardial regeneration post-infarction, but still suffer from poor retention, maturation and integration of delivered cells. Mechanical strain has been shown to alter cell size, shape, adherence and gene expression in cardiac cells. As a more recently identified cell type, the effect of mechanical strain on cardiac progenitor cells (CPCs) is unknown. This work aims to elucidate the role mechanical strain plays in CPC phenotype and if this response is matrix protein specific. We hypothesize that mechanical strain will improve CPC alignment and potential for connectivity. Methods: To examine the role of mechanical strain on CPCs, CPCs were seeded on FlexCell plates in the presence of a naturally-derived cardiac extracellularmatrix (cECM), collagen I (COL) or no protein (TCP) and strained 0% (static) or 10% at 1 Hz for 24 hours in a BioFlex system. CPC elongation, alignment, and size were evaluated by rhodamine-phalloidin staining. Connexin-43 expression was measured by Western and normalized to GAPDH. Data were analyzed by two-way ANOVA and Bonferroni post-test. Results: CPC area, independent of culture conditions, was 1020 ± 40 um2, corresponding to neonatal cardiomyocyte area. The aspect ratio (major/minor axis) of CPCs showed a trend for increased elongation with strain at (e.x. 2.0±0.2 for unstrained cECM compared to 2.7±0.1 for strained cECM; n=4, p>0.05). Static culture conditions, independent of matrix coating, showed 20±3% alignment of CPCs. Under strain, alignment increased to 30±2% on COL (n=4; p>0.05 for strained COL verus static COL) and 48±8% on cECM (n=4; p< 0.01 for strained cECM versus strained COL and p<0.001 for strained cECM verus static cECM). A fold change >2 for connexin-43 protein in strained versus static conditions, independent of matrix, was observed (n=2, p>0.05) and confirmed by immunocytochemistry. Conclusion: This work suggests that mechanical strain alters CPC phenotype. Increased strain-induced alignment appears to be matrix dependent. In conclusion, these studies provide insight into the role of both mechanical forces and biochemical responses in the function of CPCs; which could lead to improved outcomes following cellular transplantation.

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