Search results for: fermentation parameters
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 9117

Search results for: fermentation parameters

9087 Influence of Fermentation Conditions on Humic Acids Production by Trichoderma viride Using an Oil Palm Empty Fruit Bunch as the Substrate

Authors: F. L. Motta, M. H. A. Santana

Abstract:

Humic Acids (HA) were produced by a Trichoderma viride strain under submerged fermentation in a medium based on the oil palm Empty Fruit Bunch (EFB) and the main variables of the process were optimized by using response surface methodology. A temperature of 40°C and concentrations of 50g/L EFB, 5.7g/L potato peptone and 0.11g/L (NH4)2SO4 were the optimum levels of the variables that maximize the HA production, within the physicochemical and biological limits of the process. The optimized conditions led to an experimental HA concentration of 428.4±17.5 mg/L, which validated the prediction from the statistical model of 412.0mg/L. This optimization increased about 7–fold the HA production previously reported in the literature. Additionally, the time profiles of HA production and fungal growth confirmed our previous findings that HA production preferably occurs during fungal sporulation. The present study demonstrated that T. viride successfully produced HA via the submerged fermentation of EFB and the process parameters were successfully optimized using a statistics-based response surface model. To the best of our knowledge, the present work is the first report on the optimization of HA production from EFB by a biotechnological process, whose feasibility was only pointed out in previous works.

Keywords: empty fruit bunch, humic acids, submerged fermentation, Trichoderma viride

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9086 Simultaneous Saccharification and Fermentation for D-Lactic Acid Production from Dried Distillers Grains with Solubles

Authors: Nurul Aqilah Mohd Zaini, Afroditi Chatzifragkou, Dimitris Charalampopoulos

Abstract:

D-Lactic acid production is gaining increasing attention due to the thermostable properties of its polymer, Polylactic Acid (PLA). In this study, D-lactic acid was produced in microbial cultures using Lactobacillus coryniformis subsp. torquens as D-lactic acid producer and hydrolysates of Dried Distillers Grains with Solubles (DDGS) as fermentation substrate. Prior to fermentation, DDGS was first alkaline pretreated with 5% (w/v) NaOH, for 15 minutes (121oC/ ~16 psi). This led to the generation of DDGS solid residues, rich in carbohydrates and especially cellulose (~52%). The carbohydrate-rich solids were then subjected to enzymatic hydrolysis with Accellerase® 1500. For Separate Hydrolysis and Fermentation (SHF), enzymatic hydrolysis was carried out at 50oC for 24 hours, followed by fermentation of D-lactic acid at 37oC in controlled pH 6. The obtained hydrolysate contained 24 g/l glucose, 5.4 g/l xylose and 0.6 g/l arabinose. In the case of Simultaneous Saccharification and Fermentation (SSF), hydrolysis and fermentation were conducted in a single step process at 37oC in pH 5. The enzymatic hydrolysis of DGGS pretreated solids took place mostly during lag phase of L. coryniformis fermentation, with only a small amount of glucose consumed during the first 6 h. When exponential phase was started, glucose generation reduced as the microorganism started to consume glucose for D-lactic acid production. Higher concentrations of D-lactic acid were produced when SSF approach was applied, with 28 g/l D-lactic acid after 24 h of fermentation (84.5% yield). In contrast, 21.2 g/l D-lactic acid were produced when SHF was used. The optical pu rity of D-lactic acid produced from both experiments was 99.9%. Besides, approximately 2 g/l acetic acid was also generated due to lactic acid degradation after glucose depletion in SHF. SSF was proved an efficient towards DDGS ulilisation and D-lactic acid production, by reducing the overall processing time, yielding sufficient D-lactic acid concentrations without the generation of fermentation by-products.

Keywords: DDGS, alkaline pretreatment, SSF, D-lactic acid

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9085 Development of an in vitro Fermentation Chicken Ileum Microbiota Model

Authors: Bello Gonzalez, Setten Van M., Brouwer M.

Abstract:

The chicken small intestine represents a dynamic and complex organ in which the enzymatic digestion and absorption of nutrients take place. The development of an in vitro fermentation chicken small intestinal model could be used as an alternative to explore the interaction between the microbiota and nutrient metabolism and to enhance the efficacy of targeting interventions to improve animal health. In the present study we have developed an in vitro fermentation chicken ileum microbiota model for unrevealing the complex interaction of ileum microbial community under physiological conditions. A two-vessel continuous fermentation process simulating in real-time the physiological conditions of the ileum content (pH, temperature, microaerophilic/anoxic conditions, and peristaltic movements) has been standardized as a proof of concept. As inoculum, we use a pool of ileum microbial community obtained from chicken broilers at the age of day 14. The development and validation of the model provide insight into the initial characterization of the ileum microbial community and its dynamics over time-related to nutrient assimilation and fermentation. Samples can be collected at different time points and can be used to determine the microbial compositional structure, dynamics, and diversity over time. The results of studies using this in vitro model will serve as the foundation for the development of a whole small intestine in vitro fermentation chicken gastrointestinal model to complement our already established in vitro fermentation chicken caeca model. The insight gained from this model could provide us with some information about the nutritional strategies to restore and maintain chicken gut homeostasis. Moreover, the in vitro fermentation model will also allow us to study relationships between gut microbiota composition and its dynamics over time associated with nutrients, antimicrobial compounds, and disease modelling.

Keywords: broilers, in vitro model, ileum microbiota, fermentation

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9084 Biofuel Potential and Invasive Species Control: Exploring Prosopis Juliflora Pod Mash for Sustainable Energy Production

Authors: Mebrahtu Haile

Abstract:

Fuels obtained from renewable resources have garnered significant enthusiasm in recent decades due to concerns about fossil fuel depletion and climate change. This study aimed to investigate the potential of Prosopis juliflora pods mash for bio-ethanol production and its hydrolysis solid waste for solid fuel. Various parameters, such as acid concentration, hydrolysis times, fermentation times, fermentation temperature, and pH, were evaluated for their impact on bio-ethanol production using Saccharomyces cerevisiae yeast. The results showed that increasing acid concentration (up to 1 molar H₂SO₄) led to an increase in sugar content, reaching a maximum of 96.13%v/v. Optimal conditions for bio-ethanol production were found at 1 molar H₂SO₄ concentration (4.2%v/v), 48 hours fermentation time (5.1%v/v), 20 minutes hydrolysis time (5.57%v/v), 30°C fermentation temperature (5.57%v/v), and pH 5 (6.01%v/v), resulting in a maximum bio-ethanol yield of 6.01%v/v. The solid waste remaining after bio-ethanol production exhibited potential for use as a solid fuel, with a calorific value of 18.22 MJ/kg. These findings demonstrate the promising potential of Prosopis juliflora pods mash for bio-ethanol production and suggest a viable solution for addressing disposal challenges associated with solid waste, contributing to the exploration of renewable fuel sources in the face of fossil fuel depletion and climate change.

Keywords: prosopis juliflora, pods mash, invasive species, bio-ethanol, fermentation, Saccharomyces cerevisiae, solid fuel

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9083 Utilization of Extracted Spirogyra sp. Media Fermented by Gluconacetobacter Xylinum for Cellulose Production as Raw Material for Paper Product

Authors: T. S. Desak Ketut, A.n. Isna, A.a. Ayu, D. P. Ririn, Suharjono Hadiatullah

Abstract:

The requirement of paper from year to year rise rapidly. The raising of cellulose requirement in paper production caused increasing of wood requirement with the effect that limited forest areal because of deforestation. Alternative cellulose that can be used for making paper is microbial cellulose. The objective of this research are to know the effectivity fermentation media Spirogyra sp. by Gluconacetobacter xylinum for cellulose production as material for the making of paper and to know effect composition bacterial cellulose composite product of Gluconacetobacter xylinum in Spirogyra sp. The method, was used, is as follow, 1) the effect assay from variation composition of fermentation media to bacterial cellulose production by Gluconacetobacter xylinum. 2) The effect assay of composition bacterial cellulose fermentation producted by Gluconacetobacter xylinum in extracted Spirogyra media to paper quality. The result of this research is variation fermentation media Spirogyra sp. affect to production of cellulose by Gluconacetobacter xylinum. Thus, result showed by the highest value and significantly different in thickness parameter, dry weight and wet weight of nata in sucrose concentration 7,5 % and urea 0,75 %. Composition composite of bacterial cellulose from fermentation product by Gluconacetobacter xylinum in media Spirogyra sp. affect to paper quality from wet nata and dry nata. Parameters thickness, weight, water absorpsion, density and gramatur showed highest result in sucrose concentration 7,5 % and urea concentration 0,75 %, except paper density from dry nata had highest result in sucrose and urea concentration 0%.

Keywords: cellulose, fermentation media, , Gluconacetobacter xylinum, paper, Spirogyra sp.

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9082 Compositional Analysis and Antioxidant Activities of the Chocolate Fermented by Lactobacillus plantarum CK10

Authors: Hye Rim Kang, So Yae Koh, Ji-Yeon Ryu, Chang Kyu Lee, Ji Hee Lim, Hyeon A. Kim, Geun Hyung Im, Somi Kim Cho

Abstract:

In this study, antioxidant properties and compositional analysis of fermented chocolate were examined. Chocolate was fermented with Lactobacillus plantarum CK10. As fermentation time went by, pH was decreased (5.26±0.02 to 3.98±0.06) while titratable acidity was increased (5.36±0.19 to 13.31±0.34). The total polyphenol contents were maintained through the fermentation. The contents of total polyphenol were slightly increased at 8 hr (6.34±0.12 mg GAE (Gallic acid equivalent)/g), and it reached to comparable levels of the control at 24 hr (control, 5.47±0.36 mg GAE/g); 24 hr, 5.19±0.23 mg GAE/g). Similarly, the total flavonoid contents were not significantly changed during fermentation. The pronounced radical scavenging activities of chocolate, against DPPH-, ABTS-, and Alkyl radical, were observed. The levels of antioxidant activities were not dramatically altered in the course of fermentation. By gas chromatography-mass spectrometry analysis, the increase in lactic acid was measured and four major compounds, HMF, xanthosine, caffeine, and theobromine, were identified. The relative peak area of caffeine and theobromine was considerably changed during fermentation. However, no significant difference in the levels of caffeine and theobromine were observed by high-performance liquid chromatography analysis.

Keywords: antioxidant, chocolate, compositional analysis, fermentation, Lactobaillus plantarum

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9081 Isolation and Characterization of an Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation

Authors: Rukshika S Hewawasam, Sisira K. Weliwegamage, Sanath Rajapakse, Subramanium Sotheeswaran

Abstract:

Bio fuel is one of the emerging industries around the world due to arise of crisis in petroleum fuel. Fermentation is a cost effective and eco-friendly process in production of bio-fuel. So inventions in microbes, substrates, technologies in fermentation cause new modifications in fermentation. One major problem in microbial ethanol fermentation is the low resistance of conventional microorganisms to the high ethanol concentrations, which ultimately lead to decrease in the efficiency of the process. In the present investigation, an ethanol resistant bacterium was isolated from sap of Saccharum officinarum (sugar cane). The optimal cultural conditions such as pH, temperature, incubation period, and microbiological characteristics, morphological characteristics, biochemical characteristics, ethanol tolerance, sugar tolerance, growth curve assay were investigated. Isolated microorganism was tolerated to 18% (V/V) of ethanol concentration in the medium and 40% (V/V) glucose concentration in the medium. Biochemical characteristics have revealed as Gram negative, non-motile, negative for Indole test ,Methyl Red test, Voges- Proskauer`s test, Citrate Utilization test, and Urease test. Positive results for Oxidase test was shown by isolated bacterium. Sucrose, Glucose, Fructose, Maltose, Dextrose, Arabinose, Raffinose, Lactose, and Sachcharose can be utilized by this particular bacterium. It is a significant feature in effective fermentation. The fermentation process was carried out in glucose medium under optimum conditions; pH 4, temperature 30˚C, and incubated for 72 hours. Maximum ethanol production was recorded as 12.0±0.6% (V/V). Methanol was not detected in the final product of the fermentation process. This bacterium is especially useful in bio-fuel production due to high ethanol tolerance of this microorganism; it can be used to enhance the fermentation process over conventional microorganisms. Investigations are currently conducted on establishing the identity of the bacterium

Keywords: bacterium, bio-fuel, ethanol tolerance, fermentation

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9080 Optimization of Process Parameters for Peroxidase Production by Ensifer Species

Authors: Ayodeji O. Falade, Leonard V. Mabinya, Uchechukwu U. Nwodo, Anthony I. Okoh

Abstract:

Given the high utility of peroxidase in several industrial processes, the search for novel microorganisms with enhanced peroxidase production capacity is of keen interest. This study investigated the process conditions for optimum peroxidase production by Ensifer sp, new ligninolytic proteobacteria with peroxidase production potential. Also, some agricultural residues were valorized for peroxidase production under solid state fermentation. Peroxidase production was optimum at an initial medium pH 7, incubation temperature of 30 °C and agitation speed of 100 rpm using alkali lignin fermentation medium supplemented with guaiacol as the most effective inducer and ammonium sulphate as the best inorganic nitrogen. Optimum peroxidase production by Ensifer sp. was attained at 48 h with specific productivity of 12.76 ± 1.09 U mg⁻¹. Interestingly, probable laccase production was observed with optimum specific productivity of 12.76 ± 0.45 U mg⁻¹ at 72 h. The highest peroxidase yield was observed with sawdust as solid substrate under solid state fermentation. In conclusion, Ensifer sp. possesses the capacity for enhanced peroxidase production that can be exploited for various biotechnological applications.

Keywords: catalase-peroxidase, enzyme production, peroxidase, polymerase chain reaction, proteobacteria

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9079 Ergosterol Biosynthesis: Non-Conventional Method for Improving Process

Authors: Madalina Postaru, Alexandra Tucaliuc, Dan Cascaval, Anca Irina Galaction

Abstract:

Ergosterol (ergosta-5,7,22-trien-3β-ol) is the precursor of vitamin D2 (ergocalciferol), known as provitamin D2 as it is converted under UV radiation to this vitamin. The natural sources of ergosterol are mainly the yeasts (Saccharomyces sp., Candida sp.), but it can be also found in fungus (Claviceps sp.) or plants (orchids). As ergosterol is mainly accumulated in yeast cell membranes, especially in free form in the plasma-membrane, and the chemical synthesis of ergosterol does not represent an efficient method for its production, this study aimed to analyze the influence of aeration efficiency on ergosterol production by S. cerevisiae in batch and fed-batch fermentations, by considering different levels of mixing intensity, aeration rate, and n-dodecane concentration. Our previous studies on ergosterol production by S. cerevisiae in batch and fed-batch fermentation systems indicated that the addition of n-dodecane led to the increase of almost 50% of this sterol concentration, the highest productivity being reached for the fed-batch process. The experiments were carried out in a laboratory stirred bioreactor, provided with computer-controlled and recorded parameters. In batch fermentation system, the study indicated that the oxygen mass transfer coefficient, kLa, is amplified for about 3 times by increasing the volumetric concentration of n-dodecane from 0 to 15%. Moreover, the increase of dissolved oxygen concentration by adding n-dodecane leads to the diminution for 3.5 times of the produced alcohol amount. In fed-batch fermentation process, the positive influence of hydrocarbon on oxygen transfer rate is amplified mainly at its higher concentration level, as the result of the increased yeasts cells amount. Thus, by varying n-dodecane concentration from 0 to 15% vol., the kLa value increase becomes more important than for the batch fermentation, being of 4 times

Keywords: ergosterol, yeast fermentation, n-dodecane, oxygen-vector

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9078 Exploring the Biochemical and Therapeutic Properties of Aged Garlic

Authors: Farhan Saeed

Abstract:

The core objective of this work is to explicate the biochemical and therapeutic properties of aged garlic. For this purpose, two varieties of garlic were obtained from Ayub Agricultural Research Institute (AARI) Faisalabad-Pakistan. Additionally, fresh garlic was converted into aged garlic via fermentation method in the incubator at 70 to 80 % humidity level and 60C0 temperature for one month. Similarly, biochemical and antioxidant properties of fresh and aged garlic were also elucidated. Mean values showed that moisture content was decreased, whereas crude fat, crude protein, crude fiber, crude ash and total carbohydrates were enhanced after fermentation. Additionally, crude protein of fresh and aged garlic was 7.57±0.16 and 5.52±0.12%, respectively, whilst 9.68±0.41 and 8.78±0.29%, respectively, after the fermentation process. In addition, NFE contents were also enhanced up to 39% after the fermentation method. Moreover, Zn, S, Al, K, Fe, Na, Mg, and Cu contents were also increased. Furthermore, Total phenolic contents (TPC) of fresh and aged garlic were 2498.70 & 2188.50mg GAE/kg whilst 3008.59, & 2591.81mg GAE/kg for aged garlic. In conclusion, aged garlic explicated the better biochemical properties, mineral profile and antioxidant properties as compared to fresh garlic.

Keywords: aged garlic, nutritional values, bioactive properties, fermentation

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9077 Concentration of D-Pinitol from Carob Kibble Using Submerged Fermentation by Saccharomyces cerevisiae

Authors: Thi Huong Vu, Vijay Jayasena, Zhongxiang Fang, Gary Dykes

Abstract:

D-pinitol (3-O-methyl ether of D-chiro-inosito) has been known to have health benefits for diabetic patients. Carob kibble has received attention due to the presence of high value D-pinitol and polyphenol antioxidants. D-pinitol was concentrated from carob kibble using submerged fermentation with Saccharomyces cerevisiae. Total carbohydrates and D-pinitol were determined by the phenol-sulphuric acid method and HPLC, respectively. The content of D-pinitol increased from approximately 43 to 70 mg/g dry weight after fermentation. The yeast consumed over 70% of total carbohydrates in carob kibble without any negative effect on D-pinitol content. A range of substrate medium pH’s from 5.0 – 7.0 had no significant effect on the removal of carbohydrates and D-pinitol. This method may provide a practical solution for production of D-pinitol from carob in a cost effective manner.

Keywords: carob kibble, d-pinitol, saccharomyces cerevisiae, submerged fermentation, total carbohydrates

Procedia PDF Downloads 321
9076 Solid State Fermentation: A Technological Alternative for Enriching Bioavailability of Underutilized Crops

Authors: Vipin Bhandari, Anupama Singh, Kopal Gupta

Abstract:

Solid state fermentation, an eminent bioconversion technique for converting many biological substrates into a value-added product, has proven its role in the biotransformation of crops by nutritionally enriching them. Hence, an effort was made for nutritional enhancement of underutilized crops viz. barnyard millet, amaranthus and horse gram based composite flour using SSF. The grains were given pre-treatments before fermentation and these pre-treatments proved quite effective in diminishing the level of antinutrients in grains and in improving their nutritional characteristics. The present study deals with the enhancement of nutritional characteristics of underutilized crops viz. barnyard millet, amaranthus and horsegram based composite flour using solid state fermentation (SSF) as the principle bioconversion technique to convert the composite flour substrate into a nutritionally enriched value added product. Response surface methodology was used to design the experiments. The variables selected for the fermentation experiments were substrate particle size, substrate blend ratio, fermentation time, fermentation temperature and moisture content having three levels of each. Seventeen designed experiments were conducted randomly to find the effect of these variables on microbial count, reducing sugar, pH, total sugar, phytic acid and water absorption index. The data from all experiments were analyzed using Design Expert 8.0.6 and the response functions were developed using multiple regression analysis and second order models were fitted for each response. Results revealed that pretreatments proved quite handful in diminishing the level of antinutrients and thus enhancing the nutritional value of the grains appreciably, for instance, there was about 23% reduction in phytic acid levels after decortication of barnyard millet. The carbohydrate content of the decorticated barnyard millet increased to 81.5% from initial value of 65.2%. Similarly popping and puffing of horsegram and amaranthus respectively greatly reduced the trypsin inhibitor activity. Puffing of amaranthus also reduced the tannin content appreciably. Bacillus subtilis was used as the inoculating specie since it is known to produce phytases in solid state fermentation systems. These phytases remarkably reduce the phytic acid content which acts as a major antinutritional factor in food grains. Results of solid state fermentation experiments revealed that phytic acid levels reduced appreciably when fermentation was allowed to continue for 72 hours at a temperature of 35°C. Particle size and substrate blend ratio also affected the responses positively. All the parameters viz. substrate particle size, substrate blend ratio, fermentation time, fermentation temperature and moisture content affected the responses namely microbial count, reducing sugar, pH, total sugar, phytic acid and water absorption index but the effect of fermentation time was found to be most significant on all the responses. Statistical analysis resulted in the optimum conditions (particle size 355µ, substrate blend ratio 50:20:30 of barnyard millet, amaranthus and horsegram respectively, fermentation time 68 hrs, fermentation temperature 35°C and moisture content 47%) for maximum reduction in phytic acid. The model F- value was found to be highly significant at 1% level of significance in case of all the responses. Hence, second order model could be fitted to predict all the dependent parameters. The effect of fermentation time was found to be most significant as compared to other variables.

Keywords: composite flour, solid state fermentation, underutilized crops, cereals, fermentation technology, food processing

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9075 Effect of Aeration on Bacterial Cellulose (BC) Production by Gluconacetobacter xylinus DSM46604 in Batch Fermentation

Authors: Azila Adnan, Giridhar R. Nair, Mark C. Lay, Janis E. Swan

Abstract:

The effect of aeration on bacterial cellulose (BC) production by Gluconacetobacter xylinus DSM46604 was studied in 5-L bioreactor. Four aeration rates were applied (0.3, 0.6, 1.0 and 1.5 vvm) in the fermentation media at constant agitation rate, 150 rpm. One vvm enhanced BC concentration (4.4 g/L) and productivity (0.44 g/L/day) while greater agitation rate (1.5 vvm) decreased BC concentration (4.0 g/L) and productivity (0.40 g/L/day).

Keywords: aeration, bacterial cellulose, fermentation, gluconacetobacter xylinus

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9074 Biomass Production Improvement of Beauveria bassiana at Laboratory Scale for a Biopesticide Development

Authors: G. Quiroga-Cubides, M. Cruz, E. Grijalba, J. Sanabria, A. Ceballos, L. García, M. Gómez

Abstract:

Beauveria sp. has been used as an entomopathogenic microorganism for biological control of various plant pests such as whitefly, thrips, aphids and chrysomelidaes (including Cerotoma tingomariana species), which affect soybean crops in Colombia´s Altillanura region. Therefore, a biopesticide prototype based on B. bassiana strain Bv060 was developed at Corpoica laboratories. For the production of B. bassiana conidia, a baseline fermentation was performed at laboratory in a solid medium using broken rice as a substrate, a temperature of 25±2 °C and a relative humidity of 60±10%. The experimental design was completely randomized, with a three-time repetition. These culture conditions resulted in an average conidial concentration of 1.48x10^10 conidia/g, a yield of 13.07 g/kg dry substrate and a productivity of 8.83x10^7 conidia/g*h were achieved. Consequently, the objective of this study was to evaluate the influence of the particle size reduction of rice (<1 mm) and the addition of a complex nitrogen source over conidia production and efficiency parameters in a solid-state fermentation, in a completely randomized experiment with a three-time repetition. For this aim, baseline fermentation conditions of temperature and humidity were employed in a semisolid culture medium with powdered rice (10%) and a complex nitrogen source (8%). As a result, it was possible to increase conidial concentration until 9.87x10^10 conidia/g, yield to 87.07 g/g dry substrate and productivity to 3.43x10^8 conidia/g*h. This suggested that conidial concentration and yield in semisolid fermentation increased almost 7 times compared with baseline while the productivity increased 4 times. Finally, the designed system for semisolid-state fermentation allowed to achieve an easy conidia recovery, which means reduction in time and costs of the production process.

Keywords: Beauveria bassiana, biopesticide, solid state fermentation, semisolid medium culture

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9073 Improving the Quality and Nutrient Content of Palm Kernel Cake through Fermentation with Bacillus subtilis

Authors: Mirnawati, Gita Ciptaan, Ferawati

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Background and Objective: Palm kernel cake (PKC) is a waste of the palm oil industry. Indonesia, as the largest palm oil producer in the world, produced 45-46% palm kernel cake. Palm kernel cake can potentially be used as animal ration but its utilization for poultry is limited. Thus, fermentation process was done in order to increase the utilization PKC in poultry ration. An experiment was conducted to study the effect between Inoculum Doses with Bacillus subtilis and fermentation time to improve the quality and nutrient content of fermented Palm Kernel Cake. Material and Methods: 1) Palm kernel cake derived from Palm Kernel Processing Manufacture of Andalas Agro Industry in Pasaman, West Sumatra. 2) Bacillus subtilis obtained from The Research Center of Applied Chemistry LIPI, Bogor. 3) Preparations nutrient agar medium (NA) produced by Difoo - Becton Dickinson. 4) Rice bran 5) Aquades and mineral standard. The experiment used completely randomize design (CRD) with 3 x 3 factorial and 3 replications. The first factors were three doses of inoculum Bacillus subtilis: (3%), (5%), and (7%). The second factor was fermentation time: (1) 2 day, (2) 4 day, and (3) 6 day. The parameters were crude protein, crude fiber, nitrogen retention, and crude fiber digestibility of fermented palm kernel cake (FPKC). Results: The result of the study showed that there was significant interaction (P<0.01) between factor A and factor B and each factor A and B also showed significant effect (P<0.01) on crude protein, crude fiber, nitrogen retention, and crude fiber digestibility. Conclusion: From this study, it can be concluded that fermented PKC with 7% doses of Bacillus subtilis and 6 days fermentation time provides the best result as seen from 24.65% crude protein, 17.35% crude fiber, 68.47% nitrogen retention, 53.25% crude fiber digestibility of fermented palm kernel cake (FPKC).

Keywords: fermentation, Bacillus Subtilis, inoculum, palm kernel cake, quality, nutrient

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9072 Processing and Economic Analysis of Rain Tree (Samanea saman) Pods for Village Level Hydrous Bioethanol Production

Authors: Dharell B. Siano, Wendy C. Mateo, Victorino T. Taylan, Francisco D. Cuaresma

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Biofuel is one of the renewable energy sources adapted by the Philippine government in order to lessen the dependency on foreign fuel and to reduce carbon dioxide emissions. Rain tree pods were seen to be a promising source of bioethanol since it contains significant amount of fermentable sugars. The study was conducted to establish the complete procedure in processing rain tree pods for village level hydrous bioethanol production. Production processes were done for village level hydrous bioethanol production from collection, drying, storage, shredding, dilution, extraction, fermentation, and distillation. The feedstock was sundried, and moisture content was determined at a range of 20% to 26% prior to storage. Dilution ratio was 1:1.25 (1 kg of pods = 1.25 L of water) and after extraction process yielded a sugar concentration of 22 0Bx to 24 0Bx. The dilution period was three hours. After three hours of diluting the samples, the juice was extracted using extractor with a capacity of 64.10 L/hour. 150 L of rain tree pods juice was extracted and subjected to fermentation process using a village level anaerobic bioreactor. Fermentation with yeast (Saccharomyces cerevisiae) can fasten up the process, thus producing more ethanol at a shorter period of time; however, without yeast fermentation, it also produces ethanol at lower volume with slower fermentation process. Distillation of 150 L of fermented broth was done for six hours at 85 °C to 95 °C temperature (feedstock) and 74 °C to 95 °C temperature of the column head (vapor state of ethanol). The highest volume of ethanol recovered was established at with yeast fermentation at five-day duration with a value of 14.89 L and lowest actual ethanol content was found at without yeast fermentation at three-day duration having a value of 11.63 L. In general, the results suggested that rain tree pods had a very good potential as feedstock for bioethanol production. Fermentation of rain tree pods juice can be done with yeast and without yeast.

Keywords: fermentation, hydrous bioethanol, fermentation, rain tree pods, village level

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9071 Effect of Mineral Ion Addition on Yeast Performance during Very High Gravity Wort Fermentation

Authors: H. O. Udeh, T. E. Kgatla, A. I. O. Jideani

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The effect of Zn2+, Mg2+, and Ba2+ on Saccharomyces pastorianus during very high gravity fermentation was evaluated in this study at independent and three variable combinations. Wort gravity of 21oP was prepared from barley malt, hops and water, to which the metal ions were supplemented in their combinations and subsequently pitched. After 96 h of fermentation, high wort fermentability (%F)= 29.53 was obtained in wort medium containing 900:4 ppm Mg2+ + Ba2+. Increased ethanol titre 7.3491 %(v/v) and 7.1313 %(v/v) were obtained in media containing 900:4 ppm Mg2+ + Ba2+ and 12:900 ppm Zn2+ + Mg2+. Decrease %F= 22.54 and ethanol titre 6.1757% (v/v) was recorded in wort medium containing 12:4 ppm Zn2+ + Ba2+. In media containing the individual metal ions, increased %F= 27.94 and %F= 26.03 were obtained in media containing 700 ppm Mg2+ and 2 ppm Ba2+, with increased ethanol yield of 7.8844% (v/v) and 7.6245% (v/v) respectively. Least %F of 11.75 and 10.80, and ethanol titre of 4.99 (%v/v) and 4.80 (%v/v) were obtained for 10 ppm Zn2+ and 4 ppm Ba2+ respectively.

Keywords: ethanol yield, fermentability, mineral ions, yeast stress, very high gravity fermentation

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9070 Simultaneous Saccharification and Co-Fermentation of Paddy Straw and Fruit Wastes into Ethanol Production

Authors: Kamla Malik

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For ethanol production from paddy straw firstly pretreatment was done by using sodium hydroxide solution (2.0%) at 15 psi for 1 hr. The maximum lignin removal was achieved with 0.5 mm mesh size of paddy straw. It contained 72.4 % cellulose, 15.9% hemicelluloses and 2.0 % lignin after pretreatment. Paddy straw hydrolysate (PSH) with fruits wastes (5%), such as sweet lime, apple, sapota, grapes, kinnow, banana, papaya, mango, and watermelon were subjected to simultaneous saccharification and co-fermentation (SSCF) for 72 hrs by co-culture of Saccharomyces cerevisiae HAU-1 and Candida sp. with 0.3 % urea as a cheap nitrogen source. Fermentation was carried out at 35°C and determined ethanol yield at 24 hours interval. The maximum production of ethanol was produced within 72 hrs of fermentation in PSH + sapota peels (3.9% v/v) followed by PSH + kinnow peels (3.6%) and PSH+ papaya peels extract (3.1 %). In case of PSH+ banana peels and mango peel extract the ethanol produced were 2.8 % and 2.2 % (v/v). The results of this study suggest that wastes from fruits that contain fermentable sugar should not be discarded into our environment, but should be supplemented in paddy straw which converted to useful products like bio-ethanol that can serve as an alternative energy source.

Keywords: ethanol, fermentation, fruit wastes, paddy straw

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9069 Production of Biodiesel Using Tannery Fleshing as a Feedstock via Solid-State Fermentation

Authors: C. Santhana Krishnan, A. M. Mimi Sakinah, Lakhveer Singh, Zularisam A. Wahid

Abstract:

This study was initiated to evaluate and optimize the conversion of animal fat from tannery wastes into methyl ester. In the pre-treatment stage, animal fats feedstock was hydrolysed and esterified through solid state fermentation (SSF) using Microbacterium species immobilized onto sand silica matrix. After 72 hours of fermentation, predominant esters in the animal fats were found to be with 83.9% conversion rate. Later, esterified animal fats were transesterified at 3 hour reaction time with 1% NaOH (w/v %), 6% methanol to oil ratio (w/v %) to produce 89% conversion rate. C13 NMR revealed long carbon chain in fatty acid methyl esters at 22.2817-31.9727 ppm. Methyl esters of palmitic, stearic, oleic represented the major components in biodiesel.

Keywords: tannery wastes, fatty animal fleshing, trans-esterification, immobilization, solid state fermentation

Procedia PDF Downloads 267
9068 Investigation of Influence of Maize Stover Components and Urea Treatment on Dry Matter Digestibility and Fermentation Kinetics Using in vitro Gas Techniques

Authors: Anon Paserakung, Chaloemphon Muangyen, Suban Foiklang, Yanin Opatpatanakit

Abstract:

Improving nutritive values and digestibility of maize stover is an alternative way to increase their utilization in ruminant and reduce air pollution from open burning of maize stover in the northern Thailand. The present study, 2x3 factorial arrangements in completely randomized design was conducted to investigate the effect of maize stover components (whole and upper stover; cut above 5th node). Urea treatment at levels 0, 3, and 6% DM on dry matter digestibility and fermentation kinetics of maize stover using in vitro gas production. After 21 days of urea treatment, results illustrated that there was no interaction between maize stover components and urea treatment on 48h in vitro dry matter digestibility (IVDMD). IVDMD was unaffected by maize stover components (P > 0.05), average IVDMD was 55%. However, using whole maize stover gave higher cumulative gas and gas kinetic parameters than those of upper stover (P<0.05). Treating maize stover by ensiling with urea resulted in a significant linear increase in IVDMD (P<0.05). IVDMD increased from 42.6% to 53.9% when increased urea concentration from 0 to 3% and maximum IVDMD (65.1%) was observed when maize stover was ensiled with 6% urea. Maize stover treated with urea at levels of 0, 3, and 6% linearly increased cumulative gas production at 96h (31.1 vs 50.5 and 59.1 ml, respectively) and all gas kinetic parameters excepted the gas production from the immediately soluble fraction (P<0.50). The results indicate that maize stover treated with 6% urea enhance in vitro dry matter digestibility and fermentation kinetics. This study provides a practical approach to increasing utilization of maize stover in feeding ruminant animals.

Keywords: maize stover, urea treatment, ruminant feed, gas production

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9067 Effect of Acetic Acid Fermentation on Bioactive Components and Anti-Xanthine Oxidase Activities in Vinegar Brewed from Monascus-Fermented Soybeans

Authors: Kyung-Soon Choi, Ji-Young Hwang, Young-Hee Pyo

Abstract:

Vinegars have been used as an alternative remedy for treating gout, but the scientific basis remains to be elucidated. In this study, acetic acid fermentation was applied for the first time to Monascus-fermented soybeans to examine its effect on the bioactive components together with the xanthine oxidase inhibitory (XOI) activity of the soy vinegar. The content of total phenols (0.47~0.97 mg gallic acid equivalents/mL) and flavonoids (0.18~0.39 mg quercetin equivallents/mL) were spectrophotometrically determined, and the content of organic acid (10.22~59.76 mg/mL) and isoflavones (6.79~7.46 mg/mL) were determined using HPLC-UV. The analytical method for ubiquinones (0.079~0.276 μg/mL) employed saponification before solvent extraction and quantification using LC-MS. Soy vinegar also showed significant XOI (95.3%) after 20 days of acetic acid fermentation at 30 °C. The results suggest that soy vinegar has potential as a novel medicinal food.

Keywords: acetic acid fermentation, bioactive component, soy vinegar, xanthine oxidase inhibitory activity

Procedia PDF Downloads 383
9066 Cytotoxic Activity against MCF-7 Breast Cancer Cells and Antioxidant Property of Aqueous Tempe Extracts from Extended Fermentation

Authors: Zatil Athaillah, Anastasia Devi, Dian Muzdalifah, Wirasuwasti Nugrahani, Linar Udin

Abstract:

During tempe fermentation, some chemical changes occurred and they contributed to sensory, appearance, and health benefits of soybeans. Many studies on health properties of tempe have specialized on their isoflavones. In this study, other components of tempe, particularly water soluble chemicals, was investigated for their biofunctionality. The study was focused on the ability to suppress MCF-7 breast cancer cell growth and antioxidant activity, as expressed by DPPH radical scavenging activity, total phenols and total flavonoids, of the water extracts. Fermentation time of tempe was extended up to 120 hr to increase the possibility to find the functional components. Extraction yield and soluble nitrogen content were also quantified as accompanying data. Our findings suggested that yield of water extraction of tempe increased as fermentation was extended up to 120 hr, except for a slight decrease at 72 hr. Water extracts of tempe showed inhibition of MCF-7 breast cancer cell growth, as shown by lower IC50 values when compared to control (unfermented soybeans). Among the varied fermentation timescales, 60-hr period showed the highest activity (IC50 of 8.7 ± 4.95 µg/ml). The anticancer activity of extracts obtained from different fermentation time was positively correlated with total soluble nitrogens, but less relevant with antioxidant data. During 48-72 hr fermentation, at which cancer suppression activity was significant, the antioxidant properties from the three assays were not higher than control. These findings indicated that water extracts of tempe from extended fermentation could inhibit breast cancer cell growth but further study to determine the mechanism and compounds that play important role in the activity should be conducted.

Keywords: tempe, anticancer, antioxidant, phenolic compounds

Procedia PDF Downloads 245
9065 Bioproduction of Indirubin from Fermentation and Renewable Sugars Through Genomic and Metabolomic Engineering of a Bacterial Strain

Authors: Vijay H. Ingole, Efthimia Lioliou

Abstract:

Indirubin, a key bioactive component of traditional Chinese medicine, has gained increasing recognition for its potential in modern biomedical applications, particularly in pharmacology and therapeutics. The present work aimed to harness the potential by engineering an Escherichia coli strain capable of high-yield indirubin production. Through meticulous genetic engineering, we optimized the metabolic pathways in E. coli to enhance indirubin synthesis. Further, to explored the optimization of culture media and indirubin yield via batch and fed-batch fermentation techniques. By fine-tuning upstream process (USP) parameters, including nutrient composition, pH, temperature, and aeration, we established conditions that maximized both cell growth and indirubin production. Additionally, significant efforts were dedicated to refining downstream process (DSP) conditions for the extraction, purification, and quantification of indirubin. Utilizing advanced biochemical methods and analytical techniques such as UHPLC, we ensured the production of high purity indirubin. This approach not only improved the economic viability of indirubin bioproduction but also aligned with the principles of green production and sustainability.

Keywords: indirubin, bacterial strain, fermentation, HPLC

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9064 Effects of Rations with High Amount of Crude Fiber on Rumen Fermentation in Suckler Cows

Authors: H. Scholz, P. Kuehne, G. Heckenberger

Abstract:

Problems during the calving period (December until May) often are results in a high body condition score (BCS) at this time. At the end of the grazing period (frequently after early weaning), however, an increase of BCS can often be observed under German conditions. In the last eight weeks before calving, the body condition should be reduced or at least not increased. Rations with a higher amount of crude fiber can be used (rations with straw or late mowed grass silage). Fermentative digestion of fiber is slow and incomplete; that’s why the fermentative process in the rumen can be reduced over a long feeding time. Viewed in this context, feed intake of suckler cows (8 weeks before calving) in different rations and fermentation in the rumen should be checked by taking rumen fluid. Eight suckler cows (Charolais) were feeding a Total Mixed Ration (TMR) in the last eight weeks before calving and grass silage after calving. By the addition of straw (30 % [TMR1] vs. 60 % [TMR2] of dry matter) was varied the amount of crude fiber in the TMR (grass silage, straw, mineral) before calving. After calving of the cow's grass, silage [GS] was fed ad libitum, and the last measurement of rumen fluid took place on the pasture [PS]. Rumen fluid, plasma, body weight, and backfat thickness were collected. Rumen fluid pH was assessed using an electronic pH meter. Volatile fatty acids (VFA), sedimentation, methylene-blue, and amount of infusorians were measured. From these 4 parameters, an “index of rumen fermentation” [IRF] in the rumen was formed. Fixed effects of treatment (TMR1, TMR2, GS, and PS) and a number of lactations (3-7 lactations) were analyzed by ANOVA using SPSS Version 25.0 (significant by p ≤ 5 %). Rumen fluid pH was significantly influenced by variants (TMR 1 by 6.6; TMR 2 by 6.9; GS by 6.6 and PS by 6.9) but was not affected by other effects. The IRF showed disturbed fermentation in the rumen by feeding the TMR 1+2 with a high amount of crude fiber (Score: > 10.0 points) and a very good environment for fermentation during grazing the pasture (Score: 6.9 points). Furthermore, significant differences were found for VFA, methylene blue, and the number of infusorians. The use of rations with a high amount of crude fiber from weaning to calving may cause deviations from undisturbed fermentation in the rumen and adversely affect the utilization of the feed in the rumen.

Keywords: rumen fermentation, suckler cow, digestibility organic matter, crude fiber

Procedia PDF Downloads 144
9063 Device for Thermo-Magnetic Depolymerisation of Plant Biomass Prior to Methane Fermentation

Authors: Mirosław Krzemieniewski, Marcin Zieliński, Marcin Dębowski

Abstract:

This publication presents a device for depolymerisation of plant substrates applicable to agricultural biogas plants and closed-chamber sewage treatment plants where sludge fermentation is bolstered with plant mass. The device consists of a tank with a cover equipped with a heating system, an inlet for the substrate, and an outlet for the depolymerised substrate. Within the tank, a magnet shaft encased in a spiral casing is attached, equipped on its upper end with an internal magnetic disc. A motoreducer is mounted on an external magnetic disc located on the centre of the cover. Depolymerisation of the plant substrate allows for substrate destruction at much lower power levels than by conventional means. The temperature within the reactor can be lowered by 40% in comparison to existing designs. During the depolymerisation process, free radicals are generated within the magnetic field, oxidizing the conditioned substrate and promoting biodegradation. Thus, the fermentation time in the fermenters is reduced by approximately 20%.

Keywords: depolymerisation, pre-treatment, biomass, fermentation

Procedia PDF Downloads 518
9062 The Nutritive Value of Fermented Sago Pith (Metroxylon sago Rottb) Enriched with Micro Nutrients for Poultry Feed

Authors: Wizna, Helmi Muis, Hafil Abbas

Abstract:

An experiment was conducted to improve the nutrient value of sago pith (Metroxylon sago Rottb) supplemented with Zn, Sulfur and urea through fermentation by using cellulolytic bacteria (Bacillus amyloliquefaciens) as inoculums. The experiment was determination of the optimum dose combination (dosage of Zn, S and urea) for sago pith fermentation based on nutrient quality and quantity of these fermented products. The study was conducted in experimental method, using the completely randomized design in factorial with 3 treatments consist of: factor A (Dose of urea: A1 = 2.0%, A2 = 3.0%), factor B (Dose of S: B1 = 0.2%, B2 = 0.4%) and factor C (Dose of Zn: C1 = 0.0025%, C2 = 0.005%). Results of study showed that optimum condition for fermentation process of sago pith with B. amyloliquefaciens caused a change of nutrient content was obtained at urea (3%), S (0,2%), and Zn (0,0025%). This fermentation process was able to increase amino acid average, reduce crude fiber content by 67% and increase crude protein by 433%, which made the nutritional value of the product based on dry matter was 18.22% crude protein, 12.42% crude fiber, 2525 Kcal/kg metabolic energy and 65.73% nitrogen retention.

Keywords: fermentation, sago pith, sulfur, Zn, urea, Bacillus amyloliquefaciens

Procedia PDF Downloads 511
9061 Cytotoxic Activity of Acetone and Ethanol Overripe Tempe Extracts against MCF-7 Breast Cancer Cells and Their Antioxidant Property

Authors: Dian Muzdalifah, Anastasia F. Devi, Zatil A. Athaillah, Linar Z. Udin

Abstract:

Tempe is a functional food prepared from soybeans through Rhizopus spp fermentation. It is well known as functional food, originated from Indonesia. Most studies on tempe functionalities refer to ripe (48 h fermentation) tempe and only limited studies discuss overripe tempe while longer fermentation time possibly increased tempe health benefit. Hence, the present study was performed to investigate the cytotoxic activity againts MCF-7 breast cancer cells and antioxidant property of tempe prepared from 0–156 h of fermentation. Tempe samples were dried and extracted with acetone and ethanol, respectively. Their extracts were used for subsequent analysis. The cytotoxic activity was assessed on MCF 7 breast cancer cells using Alamar Blue method. The antioxidant activity was determined by DPPH free radical scavenging assay. The results indicated that acetone extracts of 108 h tempe had a potent cytotoxic activity against MCF-7 breast cancer cells (IC50 = 2.54 ± 0,30 μg/mL). Ethanol extracts of 108 h tempe also showed the potency, but at slightly higher IC50 (5.20 ± 1.01 μg/mL). Both acetone and ethanol extracts of 108 and 120 h tempe showed high antioxidant activity expressed as percent inhibition with no significant difference. However, acetone extracts of 120 h tempe (81.31 ± 3.70 %) had better ability to inhibit oxidation reaction than that of ethanol extracts (75.77 ± 6.00 %). It can be concluded that the cytotoxic activity of tempe from 0–156 h of fermentation is positively correlated to their corresponding antioxidant property. Longer fermentation time, up to 108 h, increased the ability of tempe to inhibit the growth of MCF-7 breast cancer cells and oxidative reaction. But extended fermentation time, up to 156 h, tends to decrease its ability. Further studies are encouraged to identify the active components contained in each extract.

Keywords: antioxidant property, cytotoxic activity, extracts, overripe tempeh

Procedia PDF Downloads 277
9060 Fermentation of Xylose and Glucose Mixture in Intensified Reactors by Scheffersomyces stipitis to Produce Ethanol

Authors: S. C. Santos, S. R. Dionísio, A. L. D. De Andrade, L. R. Roque, A. C. Da Costa, J. L. Ienczak

Abstract:

In this work, two fermentations at different temperatures (25 and 30 ºC), with cell recycling, were accomplished to produce ethanol, using a mix of commercial substrates, xylose (70%) and glucose (30%), as organic source for Scheffersomyces stipitis. Five consecutive fermentations of 80 g L-1 (1º, 2º and 3º recycles), 96 g L-1 (4º recycle) and 120 g L-1 (5º recycle)reduced sugars led to a final maximum ethanol concentration of 17.2 and 34.5 g L-1, at 25 and 30 ºC, respectively. Glucose was the preferred substrate; moreover xylose startup degradation was initiated after a remaining glucose presence in the medium. Results showed that yeast acid treatment, performed before each cycle, provided improvements on cell viability, accompanied by ethanol productivity of 2.16 g L-1 h-1 at 30 ºC. A maximum 36% of xylose was retained in the fermentation medium and after five-cycle fermentation an ethanol yield of 0.43 g ethanol/g sugars was observed. S. stipitis fermentation capacity and tolerance showed better results at 30 ºC with 83.4% of theoretical yield referenced on initial biomass.

Keywords: 5-carbon sugar, cell recycling fermenter, mixed sugars, xylose-fermenting yeast

Procedia PDF Downloads 417
9059 Investigation of the Variables Affecting the Use of Charcoal to Delay Fermentation in Wet Beans Slurry Using Chemical and Physical Analysis

Authors: Anuoluwapo O. Adewole

Abstract:

Fermentation is the conversion of monomeric sugars into ethanol and carbondioxide in the presence of microorganisms under anaerobic conditions. In line with the aim and objective of this research project, which is to investigate into the variables affecting the use of charcoal to delay fermentation in wet beans slurry, some physical and chemical analysis were carried out on the wet beans slurry using a PH meter in which a thermometer is incorporated in it, and a measuring cylinder was used for the foam level test. About 250 grams of the ground beans slurry was divided into two portions for testing. The sample with charcoal was labeled sample 'A' while the second sample without charcoal was labeled sample 'B' subsequently. The experiment lasted for a period of 41.15 hours (i.e., forty-one hours and nine minutes). During the fourth process, both samples could not be tested as the laboratory had been saturated with foul odor and both samples were packed and sealed in polythene bag for disposal in the trash can. It was generally observed that the sample with the charcoal lasted for a longer time before that without charcoal before total spoilage occurred.

Keywords: fermentation, monomeric sugars, beans slurry, charcoal, anaerobic conditions

Procedia PDF Downloads 332
9058 Homogenization of Cocoa Beans Fermentation to Upgrade Quality Using an Original Improved Fermenter

Authors: Aka S. Koffi, N’Goran Yao, Philippe Bastide, Denis Bruneau, Diby Kadjo

Abstract:

Cocoa beans (Theobroma cocoa L.) are the main components for chocolate manufacturing. The beans must be correctly fermented at first. Traditional process to perform the first fermentation (lactic fermentation) often consists in confining cacao beans using banana leaves or a fermentation basket, both of them leading to a poor product thermal insulation and to an inability to mix the product. Box fermenter reduces this loss by using a wood with large thickness (e>3cm), but mixing to homogenize the product is still hard to perform. Automatic fermenters are not rentable for most of producers. Heat (T>45°C) and acidity produced during the fermentation by microbiology activity of yeasts and bacteria are enabling the emergence of potential flavor and taste of future chocolate. In this study, a cylindro-rotative fermenter (FCR-V1) has been built and coconut fibers were used in its structure to confine heat. An axis of rotation (360°) has been integrated to facilitate the turning and homogenization of beans in the fermenter. This axis permits to put fermenter in a vertical position during the anaerobic alcoholic phase of fermentation, and horizontally during acetic phase to take advantage of the mid height filling. For circulation of air flow during turning in acetic phase, two woven rattan with grid have been made, one for the top and second for the bottom of the fermenter. In order to reduce air flow during acetic phase, two airtight covers are put on each grid cover. The efficiency of the turning by this kind of rotation, coupled with homogenization of the temperature, caused by the horizontal position in the acetic phase of the fermenter, contribute to having a good proportion of well-fermented beans (83.23%). In addition, beans’pH values ranged between 4.5 and 5.5. These values are ideal for enzymatic activity in the production of the aromatic compounds inside beans. The regularity of mass loss during all fermentation makes it possible to predict the drying surface corresponding to the amount being fermented.

Keywords: cocoa fermentation, fermenter, microbial activity, temperature, turning

Procedia PDF Downloads 261