Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 665

Search results for: enzymatic fermentation

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

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


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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664 Enhance Biogas Production by Enzymatic Pre-Treatment from Palm Oil Mill Effluent (POME)

Authors: M. S. Tajul Islam, Md. Zahangir Alam


To enhance biogas production through anaerobic digestion, the application of various type of pre-treatment method has some limitations in terms of sustainable environmental management. Many studies on pretreatments especially chemical and physical processes are carried out to evaluate the anaerobic digestion for enhanced biogas production. Among the pretreatment methods acid and alkali pre-treatments gained the highest importance. Previous studies have showed that although acid and alkali pretreatment has significant effect on degradation of biomass, these methods have some negative impact on environment due to their hazard in nature while enzymatic pre-treatment is environmentally friendly. One of the constrains to use of enzyme in pretreatment process for biogas production is high cost which is currently focused to reduce cost through fermentation of waste-based media. As such palm oil mill effluent (POME) as an abundant resource generated during palm oil processing at mill is being used a potential fermentation media for enzyme production. This low cost of enzyme could be an alternative to biogas pretreatment process. This review is to focus direct application of enzyme as enzymatic pre-treatment on POME to enhanced production of biogas.

Keywords: POME, enzymatic pre-treatment, biogas, lignocellulosic biomass, anaerobic digestion

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663 Effect of Different Microbial Strains on Biological Pretreatment of Sugarcane Bagasse for Enzymatic Hydrolysis

Authors: Achiraya Jiraprasertwong, Erdogan Gulari, Sumaeth Chavadej


Among agricultural residues, sugarcane bagasse is one of the most convincing raw materials for the production of bioethanol due to its availability, and low cost through enzymatic hydrolysis and yeast fermentation. A pretreatment step is needed to enhance the enzymatic step. In this study, sugarcane bagasse (SCB), one of the most abundant agricultural residues in Thailand, was pretreated biologically with various microorganisms of white-rot fungus—Phanerochaete sordid (SK 7), Cellulomonas sp. (TISTR 784), and strain A 002 (Bacillus subtilis isolated from Thai higher termites). All samples with various microbial pretreatments were further hydrolyzed enzymatically by a commercial enzyme obtained from Aspergillus niger. The results showed that the pretreatment with the white-rot fungus gave the highest glucose concentration around two-fold higher when compared with the others.

Keywords: sugarcane bagasse, microorganisms, pretreatment, enzymatic hydrolysis

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662 Enzymatic Saccharification of Dilute Alkaline Pre-treated Microalgal (Tetraselmis suecica) Biomass for Biobutanol Production

Authors: M. A. Kassim, R. Potumarthi, A. Tanksale, S. C. Srivatsa, S. Bhattacharya


Enzymatic saccharification of biomass for reducing sugar production is one of the crucial processes in biofuel production through biochemical conversion. In this study, enzymatic saccharification of dilute potassium hydroxide (KOH) pre-treated Tetraselmis suecica biomass was carried out by using cellulase enzyme obtained from Trichoderma longibrachiatum. Initially, the pre-treatment conditions were optimised by changing alkali reagent concentration, retention time for reaction, and temperature. The T. suecica biomass after pre-treatment was also characterized using Fourier Transform Infrared Spectra and Scanning Electron Microscope. These analyses revealed that the functional group such as acetyl and hydroxyl groups, structure and surface of T. suecica biomass were changed through pre-treatment, which is favourable for enzymatic saccharification process. Comparison of enzymatic saccharification of untreated and pre-treated microalgal biomass indicated that higher level of reducing sugar can be obtained from pre-treated T. suecica. Enzymatic saccharification of pre-treated T. suecica biomass was optimised by changing temperature, pH, and enzyme concentration to solid ratio ([E]/[S]). Highest conversion of carbohydrate into reducing sugar of 95% amounted to reducing sugar yield of 20 (wt%) from pre-treated T. suecica was obtained from saccharification, at temperature: 40°C, pH: 4.5 and [E]/[S] of 0.1 after 72 h of incubation. Hydrolysate obtained from enzymatic saccharification of pretreated T. suecica biomass was further fermented into biobutanol using Clostridium saccharoperbutyliticum as biocatalyst. The results from this study demonstrate a positive prospect of application of dilute alkaline pre-treatment to enhance enzymatic saccharification and biobutanol production from microalgal biomass.

Keywords: microalgal biomass, enzymatic saccharification, biobutanol, fermentation

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661 Screening and Optimization of Pretreatments for Rice Straw and Their Utilization for Bioethanol Production Using Developed Yeast Strain

Authors: Ganesh Dattatraya Saratale, Min Kyu Oh


Rice straw is one of the most abundant lignocellulosic waste materials and its annual production is about 731 Mt in the world. This study treats the subject of effective utilization of this waste biomass for biofuels production. We have showed a comparative assessment of numerous pretreatment strategies for rice straw, comprising of major physical, chemical and physicochemical methods. Among the different methods employed for pretreatment alkaline pretreatment in combination with sodium chlorite/acetic acid delignification found efficient pretreatment with significant improvement in the enzymatic digestibility of rice straw. A cellulase dose of 20 filter paper units (FPU) released a maximum 63.21 g/L of reducing sugar with 94.45% hydrolysis yield and 64.64% glucose yield from rice straw, respectively. The effects of different pretreatment methods on biomass structure and complexity were investigated by FTIR, XRD and SEM analytical techniques. Finally the enzymatic hydrolysate of rice straw was used for ethanol production using developed Saccharomyces cerevisiae SR8. The developed yeast strain enabled efficient fermentation of xylose and glucose and produced higher ethanol production. Thus development of bioethanol production from lignocellulosic waste biomass is generic, applicable methodology and have great implication for using ‘green raw materials’ and producing ‘green products’ much needed today.

Keywords: rice straw, pretreatment, enzymatic hydrolysis, FPU, Saccharomyces cerevisiae SR8, ethanol fermentation

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660 Conversion of Sweet Sorghum Bagasse to Sugars for Succinic Acid Production

Authors: Enlin Lo, Ioannis Dogaris, George Philippidis


Succinic acid is a compound used for manufacturing lacquers, resins, and other coating chemicals. It is also used in the food and beverage industry as a flavor additive. It is predominantly manufactured from petrochemicals, but it can also be produced by fermentation of sugars from renewable feedstocks, such as plant biomass. Bio-based succinic acid has great potential in becoming a platform chemical (building block) for commodity and high-value chemicals. In this study, the production of bio-based succinic acid from sweet sorghum was investigated. Sweet sorghum has high fermentable sugar content and can be cultivated in a variety of climates. In order to avoid competition with food feedstocks, its non-edible ‘bagasse’ (the fiber part after extracting the juice) was targeted. Initially, various conditions of pretreating sweet sorghum bagasse (SSB) were studied in an effort to remove most of the non-fermentable components and expose the cellulosic fiber containing the fermentable sugars (glucose). Concentrated (83%) phosphoric acid was utilized at temperatures 50-80 oC for 30-60 min at various SSB loadings (10-15%), coupled with enzymatic hydrolysis using commercial cellulase (Ctec2, Novozymes) enzyme, to identify the conditions that lead to the highest glucose yields for subsequent fermentation to succinic acid. As the pretreatment temperature and duration increased, the bagasse color changed from light brown to dark brown-black, indicating decomposition, which ranged from 15% to 72%, while the theoretical glucose yield is 91%. With Minitab software statistical analysis, a model was built to identify the optimal pretreatment condition for maximum glucose released. The projected theoretical bio-based succinic acid production is 23g per 100g of SSB, which will be confirmed with fermentation experiments using the bacterium Actinobacillus succinogenes.

Keywords: biomass, cellulose, enzymatic hydrolysis, fermentation, pretreatment, succinic acid

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659 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


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

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658 Fermentation of Wood Waste by Treating with H₃PO₄-Acetone for Bioethanol Production

Authors: Deokyeong Choe, Keonwook Nam, Young Hoon Roh


Wood waste is a potentially significant resource for economic and environment-friendly recycling. Wood waste represents a key sustainable source of biomass for transformation into bioethanol. Unfortunately, wood waste is highly recalcitrant for biotransformation, which limits its use and prevents economically viable conversion into bioethanol. As a result, an effective pretreatment is necessary to degrade cellulose of the wood waste, which improves the accessibility of cellulase. In this work, a H₃PO₄-acetone pretreatment was selected among the various pretreatment methods and used to dissolve cellulose and lignin. When the H₃PO₄ and acetone were used, 5–6% of the wood waste was found to be very appropriate for saccharification. Also, when the enzymatic saccharification was conducted in the mixture of the wood waste and 0.05 M citrate buffer solution, glucose and xylose were measured to be 80.2 g/L and 9.2 g/L respectively. Furthermore, ethanol obtained after 70 h of fermentation by S. cerevisiae was 30.4 g/L. As a result, the conversion yield from wood waste to bioethanol was calculated to be 57.4%. These results show that the pretreated wood waste can be used as good feedstocks for bioethanol production and that the H₃PO₄-acetone pretreatment can effectively increase the yield of ethanol production.

Keywords: wood waste, H₃PO₄-acetone, bioethanol, fermentation

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657 Extractive Fermentation of Ethanol Using Vacuum Fractionation Technique

Authors: Weeraya Samnuknit, Apichat Boontawan


A vacuum fractionation technique was introduced to remove ethanol from fermentation broth. The effect of initial glucose and ethanol concentrations were investigated for specific productivity. The inhibitory ethanol concentration was observed at 100 g/L. In order to increase the fermentation performance, the ethanol product was removed as soon as it is produced. The broth was boiled at 35°C by reducing the pressure to 65 mBar. The ethanol/water vapor was fractionated for up to 90 wt% before leaving the column. Ethanol concentration in the broth was kept lower than 25 g/L, thus minimized the product inhibition effect to the yeast cells. For batch extractive fermentation, a high substrate utilization rate was obtained at 26.6 g/L.h and most of glucose was consumed within 21 h. For repeated-batch extractive fermentation, addition of glucose was carried out up to 9 times and ethanol was produced more than 8-fold higher than batch fermentation.

Keywords: ethanol, extractive fermentation, product inhibition, vacuum fractionation

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656 Effects of Fermentation Techniques on the Quality of Cocoa Beans

Authors: Monday O. Ale, Adebukola A. Akintade, Olasunbo O. Orungbemi


Fermentation as an important operation in the processing of cocoa beans is now affected by the recent climate change across the globe. The major requirement for effective fermentation is the ability of the material used to retain sufficient heat for the required microbial activities. Apart from the effects of climate on the rate of heat retention, the materials used for fermentation plays an important role. Most Farmers still restrict fermentation activities to the use of traditional methods. Improving on cocoa fermentation in this era of climate change makes it necessary to work on other materials that can be suitable for cocoa fermentation. Therefore, the objective of this study was to determine the effects of fermentation techniques on the quality of cocoa beans. The materials used in this fermentation research were heap-leaves (traditional), stainless steel, plastic tin, plastic basket and wooden box. The period of fermentation varies from zero days to 10 days. Physical and chemical tests were carried out for variables in quality determination in the samples. The weight per bean varied from 1.0-1.2 g after drying across the samples and the major color of the dry beans observed was brown except with the samples from stainless steel. The moisture content varied from 5.5-7%. The mineral content and the heavy metals decreased with increase in the fermentation period. A wooden box can conclusively be used as an alternative to heap-leaves as there was no significant difference in the physical features of the samples fermented with the two methods. The use of a wooden box as an alternative for cocoa fermentation is therefore recommended for cocoa farmers.

Keywords: fermentation, effects, fermentation materials, period, quality

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655 Medium Composition for the Laboratory Production of Enzyme Fructosyltransferase (FTase)

Authors: O. R. Raimi, A. Lateef


Inoculum developments of A. niger were used for inoculation of medium for submerged fermentation and solid state fermentation. The filtrate obtained were used as sources of the extra-cellular enzymes. The FTase activities and the course of pH in submerged fermentation ranged from 7.53-24.42µ/ml and 4.4-4.8 respectively. The maximum FTase activity was obtained at 48 hours fermentation. In solid state fermentation, FTase activities ranged from 2.41-27.77µ/ml. Using ripe plantain peel and kola nut pod respectively. Both substrates supported the growth of the fungus, producing profuse growth during fermentation. In the control experiment (using kolanut pod) that lack supplementation, appreciable FTase activity of 16.92µ/ml was obtained. The optimum temperature range was 600C. it was also active at broad pH range of 1-9 with optimum obtain at pH of 5.0. FTase was stable within the range of investigated pH showing more than 60% activities. FTase can be used in the production of fructooligosaccharide, a functional food.

Keywords: Aspergillus niger, solid state fermentation, kola nut pods, Fructosyltransferase (FTase)

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654 Evaluation of Microbial Community, Biochemical and Physiological Properties of Korean Black Raspberry (Rubus coreanus Miquel) Vinegar Manufacturing Process

Authors: Nho-Eul Song, Sang-Ho Baik


Fermentation characteristics of black raspberry vinegar by using static cultures without any additives were has been investigated to establish of vinegar manufacturing conditions and improve the quality of vinegar by optimization the vinegar manufacturing process. The two vinegar manufacturing conditions were prepared; one-step fermentation condition only using mother vinegar that prepared naturally occurring black raspberry vinegar without starter yeast for alcohol fermentation (traditional method) and two-step fermentation condition using commercial wine yeast and mother vinegar for acetic acid fermentation. Approximately 12% ethanol was produced after 35 days fermentation with log 7.6 CFU/mL of yeast population in one-step fermentation, resulting sugar reduction from 14 to 6oBrix whereas in two-step fermentation, ethanol concentration was reached up to 8% after 27 days with continuous increasing yeast until log 7.0 CFU/mL. In addition, yeast and ethanol were decreased after day 60 accompanied with proliferation of acetic acid bacteria (log 5.8 CFU/mL) and titratable acidity; 4.4% in traditional method and 6% in two-step fermentation method. DGGE analysis showed that S. cerevisiae was detected until 77 days of traditional fermentation and gradually changed to AAB, Acetobacter pasteurianus, as dominant species and Komagataeibacter xylinus at the end of the fermentation. However, S. cerevisiae and A. pasteurianus was dominant in two-step fermentation process. The prepared two-step fermentation showed enhanced total polyphenol and flavonoid content significantly resulting in higher radical scavenging activity. Our studies firstly revealed the microbial community change with chemical change and demonstrated a suitable fermentation system for black raspberry vinegar by the static surface method.

Keywords: bacteria, black raspberry, vinegar fermentation, yeast

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653 Solid State Fermentation of Tamarind (Tamarindus indica) Seed to Produce Food Condiment

Authors: Olufunke O. Ezekiel, Adenike O. Ogunshe, Omotola F. Olagunju, Arinola O. Falola


Studies were conducted on fermentation of tamarind seed for production of food condiment. Fermentation followed the conventional traditional method of fermented locust bean (iru) production and was carried out over a period of three days (72 hours). Samples were withdrawn and analysed for proximate composition, pH, titratable acidity, tannin content, phytic acid content and trypsin inhibitor activity using standard methods. Effects of fermentation on proximate composition, anti-nutritional factors and sensory properties of the seed were evaluated. All data were analysed using ANOVA and means separated using Duncan multiple range test. Microbiological analysis to identify and characterize the microflora responsible for the fermentation of the seed was also carried out. Fermentation had significant effect on the proximate composition on the fermented seeds. As fermentation progressed, there was significant reduction in the anti-nutrient contents. Organisms isolated from the fermenting tamarind seeds were identified as non-pathogenic and common with fermented legumes.

Keywords: condiment, fermentation, legume, tamarind seed

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652 Analysis of Total Acid in Arabica Coffee Beans after Fermentation with Ohmic Technology

Authors: Reta


Coffee is widely consumed not only because of its typical taste, but coffee has antioxidant properties because of its polyphenols, and it stimulates brain's performance. The main problem with the consumption of coffee is its content of caffeine. Caffeine, when consumed in excess, can increase muscle tension, stimulate the heart, and increase the secretion of gastric acid. In this research, we applied ohmic-based fermentation technology, which is specially designed to mimic the stomach. We used Arabica coffee, which although cheaper than Luwak coffee, has high acidity, which needs to be reduced. Hence, we applied the ohmic technology, varied the time and temperature of the process and measured the total acidity of the coffee to determine optimum fermentation conditions. Results revealed total acidity of the coffee varied with fermentation conditions; 0.32% at 400C and 12 hr, and 0.52% at 400C and 6 hr. The longer the fermentation, the lower was the acidity. The acidity of the mongoose-fermented (natural fermentation) beans was 2.34%, which is substantially higher than the acidity of the ohmic samples. Ohmic-based fermentation technology, therefore, offers improvements in coffee quality, and this is discussed to highlight the potential of ohmic technology in coffee processing.

Keywords: ohmic technology, fermentation, coffee quality, Arabica coffee

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651 Enzymatic Esterification of Sardine Oil Processed in Morocco

Authors: M. Kharroubi, Y. Rady, F. Bellali, S. Himmi


The global objective of this study is to upgrade the sardine oil processed in Morocco by using enzymatic solutions. The specific objective of this part of study is to optimize the various parameters involved in enzymatic deacidification of fish oil processed in Morocco: pressure, ratio of oil/novozymes 435, ratio of oil/glycerol, temperature. The best deacidification yields were obtained with: -A temperature of 70 °C; -A ratio -Oil/Glycerol: 2% (% P); -A ratio -Oil/Novozyme 435: 1% (% P); -A pressure: 15 to 25 mbar. On the other hand, the study of the effect of initial oil acidity showed that whatever the acidity of the oil studied (very acidic, or low acidic), the final yields are high. Acidity does not reduce the reaction efficiency. From an industrial point of view, this represents a competitive advantage to consider. This eco-friend enzymatic solution may allows Moroccan fish oil producers to achieve acid number values that meet the standard.

Keywords: sardine oil, enzymatic esterfication, desacidification, acid number

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650 Colour Characteristics of Dried Cocoa Using Shallow Box Fermentation Technique

Authors: Khairul Bariah Sulaiman, Tajul Aris Yang


Fermentation is well known as an essential process in cocoa beans. Besides to develop the precursor of cocoa flavour, it also induce the colour changes in the beans.The fermentation process is reported to be influenced by duration of pod storage and fermentation. Therefore, this study was conducted to evaluate colour of Malaysian cocoa beans and how the pods storage and fermentation duration using shallow box technique will effect on it characteristics. There are two factors being studied ie duration of cocoa pod storage (0, 2, 4, and 6 days) and duration of cocoa fermentation (0, 1, 2, 3, 4 and 5 days). The experiment is arranged in 4 x 6 factorial design with 24 treatments and arrangement is in a Completely Randomised Design (CRD). The produced beans is inspected for colour changes under artificial light during cut test and divided into four groups of colour namely fully brown, purple brown, fully purple and slaty. Cut tests indicated that cocoa beans which are directly dried without undergone fermentation has the highest slaty percentage. However, application of pods storage before fermentation process is found to decrease the slaty percentage. In contrast, the percentages of fully brown beans start to dominate after two days of fermentation, especially from four and six days of pods storage batch. Whereas, almost all batch have percentage of fully purple less than 20%. Interestingly, the percentage of purple brown beans are scattered in the entire beans batch regardless any specific trend. Meanwhile, statistical analysis using General Linear Model showed that the pods storage has a significant effect on the colour characteristic of the Malaysian dried beans compared to fermentation duration.

Keywords: cocoa beans, colour, fermentation, shallow box

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649 Optimization of Monascus Orange Pigments Production Using pH-Controlled Fed-Batch Fermentation

Authors: Young Min Kim, Deokyeong Choe, Chul Soo Shin


Monascus pigments, commonly used as a natural colorant in Asia, have many biological activities, such as cholesterol level control, anti-obesity, anti-cancer, and anti-oxidant, that have recently been elucidated. Especially, amino acid derivatives of Monascus pigments are receiving much attention because they have higher biological activities than original Monascus pigments. Previously, there have been two ways to produce amino acid derivatives: one-step production and two-step production. However, the one-step production has low purity, and the two-step production—precursor(orange pigments) fermentation and derivatives synthesis—has low productivity and growth rate during its precursor fermentation step. In this study, it was verified that pH is a key factor that affects the stability of orange pigments and the growth rate of Monascus. With an optimal pH profile obtained by pH-stat fermentation, we designed a process of precursor(orange pigments) fermentation that is a pH-controlled fed-batch fermentation. The final concentration of orange pigments in this process increased to 5.5g/L which is about 30% higher than the concentration produced from the previously used precursor fermentation step.

Keywords: cultivation process, fed-batch fermentation, monascus pigments, pH stability

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648 Fermentation with Lactobacillus plantarum CK10 Enhanced Antioxidant Activity of Blueberry Puree

Authors: So Yae Koh, YeonWoo Song, Ji-Yeon Ryu, Jeong Yong Moon, Somi Kim Cho


Blueberry, a perennial shrub, is one of the most popular fruits due to its flavor and strong free radical scavenging properties. In this study, the blueberry puree was fermented by Lactobacillus plantarum CK10 and the antioxidant activities of fermentation products were examined. Various conditions with different supplements (5% sucrose or 10% skim milk) were evaluated for fermentation efficiency and the effects on antioxidant properties. The viable cell count of lactic acid bacteria, pH, total phenolic compounds and flavonoids contents were measured after 7 days of fermentation. DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS [2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)] radical scavenging activities were highly enhanced compared to non-fermented blueberry puree after fermentation. Interestingly, the antioxidant activities were greatly increased in the fermentation of blueberry puree alone without supplements. The present results indicate that the blueberry puree fermented by Lactobacillus plantarum CK10 could be used as a potential source of natural antioxidants and these findings will facilitate the utilization of blueberry as a resource for food additive.

Keywords: antioxidant activity, blueberry, lactobacillus plantarum CK10, fermentation

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647 High Titer Cellulosic Ethanol Production Achieved by Fed-Batch Prehydrolysis Simultaneous Enzymatic Saccharification and Fermentation of Sulfite Pretreated Softwood

Authors: Chengyu Dong, Shao-Yuan Leu


Cellulosic ethanol production from lignocellulosic biomass can reduce our reliance on fossil fuel, mitigate climate change, and stimulate rural economic development. The relative low ethanol production (60 g/L) limits the economic viable of lignocellulose-based biorefinery. The ethanol production can be increased up to 80 g/L by removing nearly all the non-cellulosic materials, while the capital of the pretreatment process increased significantly. In this study, a fed-batch prehydrolysis simultaneously saccharification and fermentation process (PSSF) was designed to converse the sulfite pretreated softwood (~30% residual lignin) to high concentrations of ethanol (80 g/L). The liquefaction time of hydrolysis process was shortened down to 24 h by employing the fed-batch strategy. Washing out the spent liquor with water could eliminate the inhibition of the pretreatment spent liquor. However, the ethanol yield of lignocellulose was reduced as the fermentable sugars were also lost during the process. Fed-batch prehydrolyzing the while slurry (i.e. liquid plus solid fraction) pretreated softwood for 24 h followed by simultaneously saccharification and fermentation process at 28 °C can generate 80 g/L ethanol production. Fed-batch strategy is very effectively to eliminate the “solid effect” of the high gravity saccharification, so concentrating the cellulose to nearly 90% by the pretreatment process is not a necessary step to get high ethanol production. Detoxification of the pretreatment spent liquor caused the loss of sugar and reduced the ethanol yield consequently. The tolerance of yeast to inhibitors was better at 28 °C, therefore, reducing the temperature of the following fermentation process is a simple and valid method to produce high ethanol production.

Keywords: cellulosic ethanol, sulfite pretreatment, Fed batch PSSF, temperature

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646 New Methodology for Monitoring Alcoholic Fermentation Processes Using Refractometry

Authors: Boukhiar Aissa, Iguergaziz Nadia, Halladj Fatima, Lamrani Yasmina, Benamara Salem


Determining the alcohol content in alcoholic fermentation bioprocess has a great importance. In fact, it is a key indicator for monitoring this fermentation bioprocess. Several methodologies (chemical, spectrophotometric, chromatographic...) are used to the determination of this parameter. However, these techniques are very long and require: rigorous preparations, sometimes dangerous chemical reagents, and/or expensive equipment. In the present study, the date juice is used as a substrate of alcoholic fermentation. The extracted juice undergoes an alcoholic fermentation by Saccharomyces cerevisiae. The study of the possible use of refractometry as a sole means for the in situ control of this process revealed a good correlation (R2 = 0.98) between initial and final ° Brix: ° Brix f = 0.377× ° Brixi. In addition, we verified the relationship between the variation in final and initial ° Brix (Δ ° Brix) and alcoholic rate produced (A exp): CΔ° Brix / A exp = 1.1. This allows the tracing of abacus isoresponses that permit to determine the alcoholic and residual sugar rates with a mean relative error (MRE) of 5.35%.

Keywords: refractometry, alcohol, residual sugar, fermentation, brix, date, juice

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645 High Efficient Biohydrogen Production from Cassava Starch Processing Wastewater by Two Stage Thermophilic Fermentation and Electrohydrogenesis

Authors: Peerawat Khongkliang, Prawit Kongjan, Tsuyoshi Imai, Poonsuk Prasertsan, Sompong O-Thong


A two-stage thermophilic fermentation and electrohydrogenesis process was used to convert cassava starch processing wastewater into hydrogen gas. Maximum hydrogen yield from fermentation stage by Thermoanaerobacterium thermosaccharolyticum PSU-2 was 248 mL H2/g-COD at optimal pH of 6.5. Optimum hydrogen production rate of 820 mL/L/d and yield of 200 mL/g COD was obtained at HRT of 2 days in fermentation stage. Cassava starch processing wastewater fermentation effluent consisted of acetic acid, butyric acid and propionic acid. The effluent from fermentation stage was used as feedstock to generate hydrogen production by microbial electrolysis cell (MECs) at an applied voltage of 0.6 V in second stage with additional 657 mL H2/g-COD was produced. Energy efficiencies based on electricity needed for the MEC were 330 % with COD removals of 95 %. The overall hydrogen yield was 800-900 mL H2/g-COD. Microbial community analysis of electrohydrogenesis by DGGE shows that exoelectrogens belong to Acidiphilium sp., Geobacter sulfurreducens and Thermincola sp. were dominated at anode. These results show two-stage thermophilic fermentation, and electrohydrogenesis process improved hydrogen production performance with high hydrogen yields, high gas production rates and high COD removal efficiency.

Keywords: cassava starch processing wastewater, biohydrogen, thermophilic fermentation, microbial electrolysis cell

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644 The Effect of Fermentation and Germination on the Nutrient and Antinutrient Composition of Lima Bean (Phaseolus lunatus) Flour

Authors: P. N. Okeke


Fermentation and germination of legumes have been an ancient practice. In this study, the influence of fermentation and germination on the chemical properties of Lima bean (Phaseolus lunatus) flour were evaluated. The flours were analyzed for their proximate and mineral composition, using the standard assay methods. The result showed that fermentation and germination increased the moisture, protein and ash content of the flours while fiber, fat and carbohydrate were decreased. The protein level of fermented and germinated lima bean increased from 21.06–26.60%. The minerals: iron, copper, zinc, and phosphorous increased due to germination and fermentation. The phytate and tannin levels were drastically reduced in both the fermented and germinated flours. The result of this study revealed that fermentation and germination makes the nutrient in lima beans more accessible as it reduces the anti-nutrients. It is therefore recommended that lima bean be process accordingly for richer and more bio-availability of the nutrients.

Keywords: nutrient, anti-nutrient, fermented, germinated, lima bean flour

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643 Kinetic Studies of Bioethanol Production from Salt-Pretreated Sugarcane Leaves

Authors: Preshanthan Moodley, E. B. Gueguim Kana


This study examines the kinetics of S. cerevisiae BY4743 growth and bioethanol production from sugarcane leaf waste (SLW), utilizing two different optimized pretreatment regimes; under two fermentation modes: steam salt-alkali filtered enzymatic hydrolysate (SSA-F), steam salt-alkali unfiltered (SSA-U), microwave salt-alkali filtered (MSA-F) and microwave salt-alkali unfiltered (MSA-U). The kinetic coefficients were determined by fitting the Monod, modified Gompertz, and logistic models to the experimental data with high coefficients of determination R² > 0.97. A maximum specific growth rate (µₘₐₓ) of 0.153 h⁻¹ was obtained under SSA-F and SSA-U whereas, 0.150 h⁻¹ was observed with MSA-F and MSA-U. SSA-U gave a potential maximum bioethanol concentration (Pₘ) of 31.06 g/L compared to 30.49, 23.26 and 21.79g/L for SSA-F, MSA-F and MSA-U respectively. An insignificant difference was observed in the μmax and Pm for the filtered and unfiltered enzymatic hydrolysate for both SSA and MSA pretreatments, thus potentially reducing a unit operation. These findings provide significant insights for process scale up.

Keywords: lignocellulosic bioethanol, microwave pretreatment, sugarcane leaves, kinetics

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642 High Dissolution of ATC by pH Control and Its Enzymatic Conversion to L-Cysteine

Authors: Deokyeong Choe, Sung Hun Youn, Younggon Kim, Chul Soo Shin


L-Cysteine is extensively used as a supplement of pharmaceuticals, cosmetics, food and feed additives. It has obtained industrially by hydrolysis of human hair and poultry feathers. However, there are some problems such as the restriction of using materials from animals and the intractable waste pollution. The enzymatic conversion has been regarded as an environmental-friendly method. Currently, the biggest bottle-neck of enzymatic conversion is the low yield of L-cysteine due to the low substrate solubility. In this study, the method of enhancing the solubility of the substrate D,L-2-amino-Δ2-thiazoline-4-carboxylicacid (ATC) was developed and the enzymatic reaction at high concentration levels was performed. A large amount of substrate in aqueous solutions was dissolved by pH control using salts. As the pH of the solution increased, the solubility of ATC increased. It was thought that a shift of ATC from acid form (-COOH) to dissociated carboxylic group (-COO-) would improve its hydrophilicity leading to solubility increase. The highest solubility of ATC was 610 mM at pH 10.5, whereas the maximum reaction rate was obtained at pH 8.3. As a result, a high L-cysteine yield of 250 mM was achieved at pH 9.1, which was obtained from a combination of optimum pH conditions for ATC solubility and enzymatic conversion. This yield corresponds to approximately 18 times of that in previous reports.

Keywords: D, L-2-amino-Δ2-thiazoline-4-carboxylicacid, enzymatic conversion, high-substrate solubilization, L-Cysteine

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641 Mathematical Modeling of Bi-Substrate Enzymatic Reactions in the Presence of Different Types of Inhibitors

Authors: Rafayel Azizyan, Valeri Arakelyan, Aram Gevorgyan, Varduhi Balayan, Emil Gevorgyan


Currently, mathematical and computer modeling are widely used in different biological studies to predict or assess behavior of such complex systems as biological ones. This study deals with mathematical and computer modeling of bi-substrate enzymatic reactions, which play an important role in different biochemical pathways. The main objective of this study is to represent the results from in silico investigation of bi-substrate enzymatic reactions in the presence of uncompetitive inhibitors, as well as to describe in details the inhibition effects. Four models of uncompetitive inhibition were designed using different software packages. Particularly, uncompetitive inhibitor to the first [ES1] and the second ([ES1S2]; [FS2]) enzyme-substrate complexes have been studied. The simulation, using the same kinetic parameters for all models allowed investigating the behavior of reactions as well as determined some interesting aspects concerning influence of different cases of uncompetitive inhibition. Besides that shown, that uncompetitive inhibitors exhibit specific selectivity depending on mechanism of bi-substrate enzymatic reaction.

Keywords: mathematical modeling, bi-substrate enzymatic reactions, reversible inhibition

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640 Traditional Dyeing of Silk with Natural Dyes by Eco-Friendly Method

Authors: Samera Salimpour Abkenar


In traditional dyeing of natural fibers with natural dyes, metal salts are commonly used to increase color stability. This method always carries the risk of environmental pollution (contamination of arable soils and fresh groundwater) due to the release of dyeing effluents containing large amounts of metal. Therefore, researchers are always looking for new methods to obtain a green dyeing system. In this research, the use of the enzymatic dyeing method to prevent environmental pollution with metals and reduce production costs has been proposed. After degumming and bleaching, raw silk fabrics were dyed with natural dyes (Madder and Sumac) by three methods (pre-mordanting with a metal salt, one-step enzymatic dyeing, and two-step enzymatic dyeing). Results show that silk dyed with natural dyes by the enzymatic method has higher color strength and colorfastness than the pretreated with a metal salt. Also, the amount of remained dyes in the dyeing wastewater is significantly reduced by the enzymatic method. It is found that the enzymatic dyeing method leads to improvement of dye absorption, color strength, soft hand, no change in color shade, low production costs (due to low dyeing temperature), and a significant reduction in environmental pollution.

Keywords: eco-friendly, natural dyes, silk, traditional dyeing

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639 Assessment of Green Fluorescent Protein Signal for Effective Monitoring of Recombinant Fermentation Processes

Authors: I. Sani, A. Abdulhamid, F. Bello, Isah M. Fakai


This research has focused on the application of green fluorescent protein (GFP) as a new technique for direct monitoring of fermentation processes involving cultured bacteria. To use GFP as a sensor for pH and oxygen, percentage ratio of red fluorescence to green (% R/G) was evaluated. Assessing the magnitude of the % R/G ratio in relation to low or high pH and oxygen concentration, the bacterial strains were cultivated under aerobic and anaerobic conditions. SCC1 strains of E. coli were grown in a 5 L laboratory fermenter, and during the fermentation, the pH and temperature were controlled at 7.0 and 370C respectively. Dissolved oxygen tension (DOT) was controlled between 15-100% by changing the agitation speed between 20-500 rpm respectively. Effect of reducing the DOT level from 100% to 15% was observed after 4.5 h fermentation. There was a growth arrest as indicated by the decrease in the OD650 at this time (4.5-5 h). The relative fluorescence (green) intensity was decreased from about 460 to 420 RFU. However, %R/G ratio was significantly increased from about 0.1% to about 0.25% when the DOT level was decreased to 15%. But when the DOT was changed to 100%, a little increase in the RF and decrease in the %R/G ratio were observed. Therefore, GFP can effectively detect and indicate any change in pH and oxygen level during fermentation processes.

Keywords: Escherichia coli SCC1, fermentation process, green fluorescent protein, red fluorescence

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638 Optimization of Digestive Conditions of Opuntia ficus-indica var. Saboten using Food-Grade Enzymes

Authors: Byung Wook Yang, Sae Kyul Kim, Seung Il Ahn, Jae Hee Choi, Heejung Jung, Yejin Choi, Byung Yong Kim, Young Tae Hahm


Opuntia ficus-indica is a member of the Cactaceae family that is widely grown in all the semiarid countries throughout the world. Opuntia ficus-indica var. Saboten (OFS), commonly known as prickly pear cactus, is commercially cultivated as a dietary foodstuffs and medicinal stuffs in Jeju Island, Korea. Owing to high viscosity of OFS’ pad, its application to the commercial field has been limited. When the low viscosity of OFS’s pad is obtained, it is useful for the manufacture of healthy food in the related field. This study was performed to obtain the optimal digestion conditions of food-grade enzymes (Pectinex, Viscozyme and Celluclast) with the powder of OFS stem. And also, the contents of water-soluble dietary fiber (WSDF) of the dried powder prepared by the extraction of OFS stem were monitored and optimized using the response surface methodology (RSM), which included 20 experimental points with 3 replicates for two independent variables (fermentation temperature and time). A central composite design was used to monitor the effect of fermentation temperature (30-90 °C, X1) and fermentation time (1-10h, X2) on dependent variables, such as viscosity (Y1), water-soluble dietary fiber (Y2) and dietary fiber yield (Y3). Estimated maximum values at predicted optimum conditions were in agreement with experimental values. Optimum temperature and duration were 50°C and 12 hours, respectively. Viscosity value reached 3.4 poise. Yield of water-soluble dietary fiber is determined in progress.

Keywords: Opuntia ficus-indica var. saboten, enzymatic fermentation, response surface methodology, water-soluble dietary fiber, viscosity

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637 Effect of Fermentation Time on Some Functional Properties of Moringa (Moringa oleifera) Seed Flour

Authors: Ocheme B. Ocheme, Omobolanle O. Oloyede, S. James, Eleojo V. Akpa


The effect of fermentation time on some functional properties of Moringa (Moringa oleifera) seed flour was examined. Fermentation, an effective processing method used to improve nutritional quality of plant foods, tends to affect the characteristics of food components and their behaviour in food systems just like other processing methods. Hence the need for this study. Moringa seeds were fermented naturally by soaking in potable water and allowing it to stand for 12, 24, 48 and 72 hours. At the end of fermentation, the seeds were oven dried at 600C for 12 hours and then milled into flour. Flour obtained from unfermented seeds served as control: hence a total of five flour samples. The functional properties were analyzed using standard methods. Fermentation significantly (p<0.05) increased the water holding capacity of Moringa seed flour from 0.86g/g - 2.31g/g. The highest value was observed after 48 hours of fermentation The same trend was observed for oil absorption capacity with values between 0.87 and 1.91g/g. Flour from unfermented Moringa seeds had a bulk density of 0.60g/cm3 which was significantly (p<0.05) higher than the bulk densities of flours from seeds fermented for 12, 24 and 48. Fermentation significantly (p<0.05) decreased the dispersibility of Moringa seed flours from 36% to 21, 24, 29 and 20% after 12, 24, 48 and 72 hours of fermentation respectively. The flours’ emulsifying capacities increased significantly (p<0.05) with increasing fermentation time with values between 50 – 68%. The flour obtained from seeds fermented for 12 hours had a significantly (p<0.05) higher foaming capacity of 16% while the flour obtained from seeds fermented for 0, 24 and 72 hours had the least foaming capacities of 9%. Flours from seeds fermented for 12 and 48 hours had better functional properties than flours from seeds fermented for 24 and 72 hours.

Keywords: fermentation, flour, functional properties, Moringa

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636 Application of Refractometric Methodology for Simultaneous Determination of Alcohol and Residual Sugar Concentrations during Alcoholic Fermentation Bioprocess of Date Juice

Authors: Boukhiar Aissa, Halladj Fatima, Iguergaziz Nadia, Lamrani yasmina, Benamara Salem


Determining the alcohol content in alcoholic fermentation bioprocess is of great importance. In fact, it is a key indicator for monitoring this bioprocess. Several methodologies (chemical, spectrophotometric, chromatographic) are used to the determination of this parameter. However, these techniques are very long and they require: rigorous preparations, sometimes dangerous chemical reagents and/or expensive equipment. In the present study, the date juice is used as the substrate of alcoholic fermentation. The extracted juice undergoes an alcoholic fermentation by Saccharomyces cerevisiae. The study of the possible use of refractometry as a sole means for the in situ control of alcoholic fermentation revealed a good correlation (R2=0.98) between initial and final °Brix: °Brixf=0.377×°Brixi. In addition, the relationship between Δ°Brix and alcoholic content of the final product (A,%) has been determined: Δ°Brix/A=1.1. The obtained results allowed us to establish iso-responses abacus, which can be used for the determination of alcohol and residual sugar content, with a mean relative error (MRE) of 5.35%.

Keywords: alcoholic fermentation, date juice, refractometry, residual sugar

Procedia PDF Downloads 247