Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1872

Search results for: liquid fermentation

1872 Isolation, Identification and Characterization of the Bacteria and Yeast from the Fermented Stevia Extract

Authors: Asato Takaishi, Masashi Nasuhara, Ayuko Itsuki, Kenichi Suga


Stevia (Stevia rebaudiana Bertoni) is a composite plant native to Paraguay. Stevia sweetener is derived from a hot water extract of Stevia (Stevia extract), which has some effects such as histamine decomposition, antioxidative effect, and blood sugar level-lowering function. The steviol glycosides in the Stevia extract are considered to contribute to these effects. In addition, these effects increase by the fermentation. However, it takes a long time for fermentation of Stevia extract and the fermentation liquid sometimes decays during the fermentation process because natural fermentation method is used. The aim of this study is to perform the fermentation of Stevia extract in a shorter period, and to produce the fermentation liquid in stable quality. From the natural fermentation liquid of Stevia extract, the four strains of useful (good taste) microorganisms were isolated using dilution plate count method and some properties were determined. The base sequences of 16S rDNA and 28S rDNA revealed three bacteria (two Lactobacillus sp. and Microbacterium sp.) and one yeast (Issatchenkia sp.). This result has corresponded that several kinds of lactic bacterium such as Lactobacillus pentosus and Lactobacillus buchneri were isolated from Stevia leaves. Liquid chromatography/mass spectrometory (LC/MS/MS) and High-Performance Liquid Chromatography (HPLC) were used to determine the contents of steviol glycosides and neutral sugars. When these strains were cultured in the sterile Stevia extract, the steviol and stevioside were increased in the fermented Stevia extract. So, it was suggested that the rebaudioside A and the mixture of steviol glycosides in the Stevia extract were decomposed into stevioside and steviol by microbial metabolism.

Keywords: fermentation, lactobacillus, Stevia, steviol glycosides, yeast

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1871 Adjustment and Scale-Up Strategy of Pilot Liquid Fermentation Process of Azotobacter sp.

Authors: G. Quiroga-Cubides, A. Díaz, M. Gómez


The genus Azotobacter has been widely used as bio-fertilizer due to its significant effects on the stimulation and promotion of plant growth in various agricultural species of commercial interest. In order to obtain significantly viable cellular concentration, a scale-up strategy for a liquid fermentation process (SmF) with two strains of A. chroococcum (named Ac1 and Ac10) was validated and adjusted at laboratory and pilot scale. A batch fermentation process under previously defined conditions was carried out on a biorreactor Infors®, model Minifors of 3.5 L, which served as a baseline for this research. For the purpose of increasing process efficiency, the effect of the reduction of stirring speed was evaluated in combination with a fed-batch-type fermentation laboratory scale. To reproduce the efficiency parameters obtained, a scale-up strategy with geometric and fluid dynamic behavior similarities was evaluated. According to the analysis of variance, this scale-up strategy did not have significant effect on cellular concentration and in laboratory and pilot fermentations (Tukey, p > 0.05). Regarding air consumption, fermentation process at pilot scale showed a reduction of 23% versus the baseline. The percentage of reduction related to energy consumption reduction under laboratory and pilot scale conditions was 96.9% compared with baseline.

Keywords: Azotobacter chroococcum, scale-up, liquid fermentation, fed-batch process

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1870 Evaluation of Liquid Fermentation Strategies to Obtain a Biofertilizer Based on Rhizobium sp.

Authors: Andres Diaz Garcia, Ana Maria Ceballos Rojas, Duvan Albeiro Millan Montano


This paper describes the initial technological development stages in the area of liquid fermentation required to reach the quantities of biomass of the biofertilizer microorganism Rhizobium sp. strain B02, for the application of the unitary stages downstream at laboratory scale. In the first stage, the adjustment and standardization of the fermentation process in conventional batch mode were carried out. In the second stage, various fed-batch and continuous fermentation strategies were evaluated in 10L-bioreactor in order to optimize the yields in concentration (Colony Forming Units/ml•h) and biomass (g/l•h), to make feasible the application of unit operations downstream of process. The growth kinetics, the evolution of dissolved oxygen and the pH profile generated in each of the strategies were monitored and used to make sequential adjustments. Once the fermentation was finished, the final concentration and viability of the obtained biomass were determined and performance parameters were calculated with the purpose of select the optimal operating conditions that significantly improved the baseline results. Under the conditions adjusted and standardized in batch mode, concentrations of 6.67E9 CFU/ml were reached after 27 hours of fermentation and a subsequent noticeable decrease was observed associated with a basification of the culture medium. By applying fed-batch and continuous strategies, significant increases in yields were achieved, but with similar concentration levels, which involved the design of several production scenarios based on the availability of equipment usage time and volume of required batch.

Keywords: biofertilizer, liquid fermentation, Rhizobium sp., standardization of processes

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1869 Bioproduction of Phytohormones by Liquid Fermentation Using a Mexican Strain of Botryodiplodia theobromae

Authors: Laredo Alcalá Elan Iñaky, Hernandez Castillo Daniel, Martinez Hernandez José Luis, Arredondo Valdes Roberto, Gonzalez Gallegos Esmeralda, Anguiano Cabello Julia Cecilia


Plant hormones are a group of molecules that control different processes ranging from the growth and development of the plant until their response to biotic and abiotic stresses. In this study, the capacity of production of various phytohormones was evaluated from a strain of Botryodiplodia theobromae by liquid fermentation system using the modified Mierch medium added with a hydrolyzate compound of mead all in a reactor without agitation at 28 °C for 15 days. Quantification of the metabolites was performed using high performance liquid chromatography techniques. The results showed that a microbial broth with at least five different types of plant hormones was obtained: gibberellic acid, zeatin, kinetin, indoleacetic acid and jasmonic acid, the last one was higher than the others metabolites produced. The production of such hormones using a single type of microorganism could be in the future a great alternative to reduce production costs and similarly reduce the use of synthetic chemicals.

Keywords: biosystem, plant hormones, Botryodiplodia theobromae, fermentation

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1868 Comparison of Filamentous Fungus (Monascus purpureus)Growth in Submerged and Solid State Culture

Authors: Shafieeh Mansoori, Fatemeh Yazdian, Ashrafsadat Hatamian, Majid Azizi


Monascus purpureus, which has a special metabolite with many therapeutic and medicinal properties including antioxidant, antibiotic, anti-hypercholesterolemia, and immunosuppressive properties, is a traditional Chinese fermentation fungus and is used as a natural dietary supplement. Production of desired metabolites actually determined by optimized growth which is supported by some factors such as substrates and Monascus strains type, moisture content of the fermentation mixture, aeration, and control of contamination issues. In this experiment, M. purpureus PTCC5305 was cultured in both the liquid and solid culture medium. The former medium contain YMP (yeast extract, maltose and peptone), PGC (peptone, glucose complex), and GYP (glucose, yeast extract and peptone) medium. After 8 days, the best medium for the cell production was PGC agar medium on solid culture with 0.28 g dry weight of cell mass whereas the best liquid culture was GYP medium with 3.5 g/l dry weight of cell mass. The lowest cell production was on YMP agar with 0.1 g dry weight of cell mass and then YMP medium with 2.5 g/l dry cell weight.

Keywords: Monascus purpureus, solid state fermentation, submerged culture, Chinese fermentation fungus

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1867 Usage of Crude Glycerol for Biological Hydrogen Production, Experiments and Analysis

Authors: Ilze Dimanta, Zane Rutkovska, Vizma Nikolajeva, Janis Kleperis, Indrikis Muiznieks


Majority of word’s steadily increasing energy consumption is provided by non-renewable fossil resources. Need to find an alternative energy resource is essential for further socio-economic development. Hydrogen is renewable, clean energy carrier with high energy density (142 MJ/kg, accordingly – oil has 42 MJ/kg). Biological hydrogen production is an alternative way to produce hydrogen from renewable resources, e.g. using organic waste material resource fermentation that facilitate recycling of sewage and are environmentally benign. Hydrogen gas is produced during the fermentation process of bacteria in anaerobic conditions. Bacteria are producing hydrogen in the liquid phase and when thermodynamic equilibrium is reached, hydrogen is diffusing from liquid to gaseous phase. Because of large quantities of available crude glycerol and the highly reduced nature of carbon in glycerol per se, microbial conversion of it seems to be economically and environmentally viable possibility. Such industrial organic waste product as crude glycerol is perspective for usage in feedstock for hydrogen producing bacteria. The process of biodiesel production results in 41% (w/w) of crude glycerol. The developed lab-scale test system (experimental bioreactor) with hydrogen micro-electrode (Unisense, Denmark) was used to determine hydrogen production yield and rate in the liquid phase. For hydrogen analysis in the gas phase the RGAPro-100 mass-spectrometer connected to the experimental test-system was used. Fermentative bacteria strains were tested for hydrogen gas production rates. The presence of hydrogen in gaseous phase was measured using mass spectrometer but registered concentrations were comparatively small. To decrease the hydrogen partial pressure in liquid phase reactor with a system for continuous bubbling with inert gas was developed. H2 production rate for the best producer in liquid phase reached 0,40 mmol H2/l, in gaseous phase - 1,32 mmol H2/l. Hydrogen production rate is time dependent – higher rate of hydrogen production is at the fermentation process beginning when concentration increases, but after three hours of fermentation, it decreases.

Keywords: bio-hydrogen, fermentation, experimental bioreactor, crude glycerol

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1866 Schizosaccharomyces pombe, Saccharomyces cerevisiae Yeasts and Acetic Acid Bacteria in Alcoholic and Acetous Fermentations: Effect on Phenolic Acids of Kei-Apple (Dovyalis caffra L.) Vinegar

Authors: Phillip Minnaar, Neil Jolly, Louisa Beukes, Santiago Benito-Saez


Dovyalis caffra is a tree found on the African continent. Limited information exists on the effect of acetous fermentation on the phytochemicals of Kei-apple fruit. The phytochemical content of vinegars is derived from compounds present in the fruit the vinegar is made of. Kei-apple fruit juice was co-inoculated with Schizosaccharomyces pombe and Saccharomyces cerevisiae to induce alcoholic fermentation (AF). Acetous fermentation followed AF, using an acetic acid bacteria consortium as an inoculant. Juice had the lowest pH and highest total acidity (TA). The wine had the highest pH and vinegars lowest TA. Total soluble solids and L-malic acid decreased during AF and acetous fermentation. Volatile acidity concentration was not different among vinegars. Gallic, syringic, caffeic, p-coumaric, and chlorogenic acids increased during acetous fermentation, whereas ferulic, sinapic, and protocatechuic acids decreased. Chlorogenic acid was the most abundant phenolic acid in both wines and vinegars. It is evident from this investigation that Kei-apple vinegar is a source of plant-derived phenolics, which evolved through fermentation. However, the AAB selection showed minimal performance with respect to VA production. Acetic acid bacteria selection for acetous fermentation should be reconsidered, and the reasons for the decrease of certain phenolic acids during acetous fermentation needs to be investigated.

Keywords: acetic acid bacteria, acetous fermentation, liquid chromatography, phenolic acids

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1865 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


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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1864 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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1863 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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1862 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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1861 Fermentation of Pretreated Herbaceous Cellulosic Wastes to Ethanol by Anaerobic Cellulolytic and Saccharolytic Thermophilic Clostridia

Authors: Lali Kutateladze, Tamar Urushadze, Tamar Dudauri, Besarion Metreveli, Nino Zakariashvili, Izolda Khokhashvili, Maya Jobava


Lignocellulosic waste streams from agriculture, paper and wood industry are renewable, plentiful and low-cost raw materials that can be used for large-scale production of liquid and gaseous biofuels. As opposed to prevailing multi-stage biotechnological processes developed for bioconversion of cellulosic substrates to ethanol where high-cost cellulase preparations are used, Consolidated Bioprocessing (CBP) offers to accomplish cellulose and xylan hydrolysis followed by fermentation of both C6 and C5 sugars to ethanol in a single-stage process. Syntrophic microbial consortium comprising of anaerobic, thermophilic, cellulolytic, and saccharolytic bacteria in the genus Clostridia with improved ethanol productivity and high tolerance to fermentation end-products had been proposed for achieving CBP. 65 new strains of anaerobic thermophilic cellulolytic and saccharolytic Clostridia were isolated from different wetlands and hot springs in Georgia. Using new isolates, fermentation of mechanically pretreated wheat straw and corn stalks was done under oxygen-free nitrogen environment in thermophilic conditions (T=550C) and pH 7.1. Process duration was 120 hours. Liquid and gaseous products of fermentation were analyzed on a daily basis using Perkin-Elmer gas chromatographs with flame ionization and thermal detectors. Residual cellulose, xylan, xylose, and glucose were determined using standard methods. Cellulolytic and saccharolytic bacteria strains degraded mechanically pretreated herbaceous cellulosic wastes and fermented glucose and xylose to ethanol, acetic acid and gaseous products like hydrogen and CO2. Specifically, maximum yield of ethanol was reached at 96 h of fermentation and varied between 2.9 – 3.2 g/ 10 g of substrate. The content of acetic acid didn’t exceed 0.35 g/l. Other volatile fatty acids were detected in trace quantities.

Keywords: anaerobic bacteria, cellulosic wastes, Clostridia sp, ethanol

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1860 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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1859 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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1858 Production and Purification of Monosaccharides by Hydrolysis of Sugar Cane Bagasse in an Ionic Liquid Medium

Authors: T. R. Bandara, H. Jaelani, G. J. Griffin


The conversion of lignocellulosic waste materials, such as sugar cane bagasse, to biofuels such as ethanol has attracted significant interest as a potential element for transforming transport fuel supplies to totally renewable sources. However, the refractory nature of the cellulosic structure of lignocellulosic materials has impeded progress on developing an economic process, whereby the cellulose component may be effectively broken down to glucose monosaccharides and then purified to allow downstream fermentation. Ionic liquid (IL) treatment of lignocellulosic biomass has been shown to disrupt the crystalline structure of cellulose thus potentially enabling the cellulose to be more readily hydrolysed to monosaccharides. Furthermore, conventional hydrolysis of lignocellulosic materials yields byproducts that are inhibitors for efficient fermentation of the monosaccharides. However, selective extraction of monosaccharides from an aqueous/IL phase into an organic phase utilizing a combination of boronic acids and quaternary amines has shown promise as a purification process. Hydrolysis of sugar cane bagasse immersed in an aqueous solution with IL (1-ethyl-3-methylimidazolium acetate) was conducted at different pH and temperature below 100 ºC. It was found that the use of a high concentration of hydrochloric acid to acidify the solution inhibited the hydrolysis of bagasse. At high pH (i.e. basic conditions), using sodium hydroxide, catalyst yields were reduced for total reducing sugars (TRS) due to the rapid degradation of the sugars formed. For purification trials, a supported liquid membrane (SLM) apparatus was constructed, whereby a synthetic solution containing xylose and glucose in an aqueous IL phase was transported across a membrane impregnated with phenyl boronic acid/Aliquat 336 to an aqueous phase. The transport rate of xylose was generally higher than that of glucose indicating that a SLM scheme may not only be useful for purifying sugars from undesirable toxic compounds, but also for fractionating sugars to improve fermentation efficiency.

Keywords: biomass, bagasse, hydrolysis, monosaccharide, supported liquid membrane, purification

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1857 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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1856 Stability of Ochratoxin a During Bread Making Process

Authors: Sara Heidari, Jafar Mohammadzadeh Milani, Elmira Pouladi Borj


In this research, stability of Ochratoxin A (OTA) during bread making process including fermentation with yeasts (Saccharomyces cerevisiae) and Sourdough (Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus and Lactobacillus fermentum) and baking at 200°C were examined. Bread was prepared on a pilot-plant scale by using wheat flour spiked with standard solution of OTA. During this process, mycotoxin levels were determined after fermentation of the dough with sourdough and three types of yeast including active dry yeast, instant dry yeast and compressed yeast after further baking 200°C by high performance liquid chromatography (HPLC) with fluorescence detector after extraction and clean-up on an immunoaffinity column. According to the results, the highest stability of was observed in the first fermentation (first proof), while the lowest stability was observed in the baking stage in comparison to contaminated flour. In addition, compressed yeast showed the maximum impact on stability of OTA during bread making process.

Keywords: Ochratoxin A, bread, dough, yeast, sourdough

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1855 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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1854 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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1853 Optimization of Fermentation Parameters for Bioethanol Production from Waste Glycerol by Microwave Induced Mutant Escherichia coli EC-MW (ATCC 11105)

Authors: Refal Hussain, Saifuddin M. Nomanbhay


Glycerol is a valuable raw material for the production of industrially useful metabolites. Among many promising applications for the use of glycerol is its bioconversion to high value-added compounds, such as bioethanol through microbial fermentation. Bioethanol is an important industrial chemical with emerging potential as a biofuel to replace vanishing fossil fuels. The yield of liquid fuel in this process was greatly influenced by various parameters viz, temperature, pH, glycerol concentration, organic concentration, and agitation speed were considered. The present study was undertaken to investigate optimum parameters for bioethanol production from raw glycerol by immobilized mutant Escherichia coli (E.coli) (ATCC11505) strain on chitosan cross linked glutaraldehyde optimized by Taguchi statistical method in shake flasks. The initial parameters were set each at four levels and the orthogonal array layout of L16 (45) conducted. The important controlling parameters for optimized the operational fermentation was temperature 38 °C, medium pH 6.5, initial glycerol concentration (250 g/l), and organic source concentration (5 g/l). Fermentation with optimized parameters was carried out in a custom fabricated shake flask. The predicted value of bioethanol production under optimized conditions was (118.13 g/l). Immobilized cells are mainly used for economic benefits of continuous production or repeated use in continuous as well as in batch mode.

Keywords: bioethanol, Escherichia coli, immobilization, optimization

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1852 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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1851 Numerical Simulation and Analysis on Liquid Nitrogen Spray Heat Exchanger

Authors: Wenjing Ding, Weiwei Shan, Zijuan, Wang, Chao He


Liquid spray heat exchanger is the critical equipment of temperature regulating system by gaseous nitrogen which realizes the environment temperature in the range of -180 ℃~+180 ℃. Liquid nitrogen is atomized into smaller liquid drops through liquid nitrogen sprayer and then contacts with gaseous nitrogen to be cooled. By adjusting the pressure of liquid nitrogen and gaseous nitrogen, the flowrate of liquid nitrogen is changed to realize the required outlet temperature of heat exchanger. The temperature accuracy of shrouds is ±1 ℃. Liquid nitrogen spray heat exchanger is simulated by CATIA, and the numerical simulation is performed by FLUENT. The comparison between the tests and numerical simulation is conducted. Moreover, the results help to improve the design of liquid nitrogen spray heat exchanger.

Keywords: liquid nitrogen spray, temperature regulating system, heat exchanger, numerical simulation

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1850 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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1849 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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1848 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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1847 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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1846 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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1845 Effect of Alcoholic and Acetous Fermentations on Phenolic Acids of Kei-Apple (Dovyalis Caffra L.) Fruit

Authors: Neil Jolly, Louisa Beukes, Santiago Benito-SaEz


Kei-apple is a tree found on the African continent. Limited information exists on the effect of alcoholic and acetous fermentation on the phytochemicals. The fruit has increased L-malic, ascorbic, and phenolic acids. Juice was co-inoculated with Schizosaccharomyces pombe and Saccharomyces cerevisiae to induce alcoholic fermentation and acetous fermentation using acetic acid bacteria. Saccharomyces cerevisiae+S. pombe wines and vinegars had highest pH. Total acidity, soluble solids and L-malic acid decreased during alcoholic and acetous fermentation with highest in S. cerevisiae wines and vinegars. Volatile acidity was highest in S. pombe vinegars but not different from S. cerevisiae and S. cerevisiae+S. pombe. Gallic acid was highest in S. pombe wines and vinegars. Syringic acid was highest in S. cerevisiae wines and vinegars. S. cerevisiae+S. pombe wines were highest in caffeic, p-coumaric and protocatechuic acids. Schizosaccharomyces pombe vinegars were highest in caffeic and p-coumaric acids. Ferulic and sinapic acids were highest in S. pombe and S. cerevisiae wines, respectively. Chlorogenic acid was most abundant in both wines and vinegars. Saccharomyces cerevisiae+S. pombe and S. cerevisiae had a positive effect on most phenolic acids. Saccharomyces cerevisiae +acetic acid bacteria had an increased effect on syringic and chlorogenic acids. Schizosaccharomyces pombe+acetic acid bacteria resulted in an increase in gallic, caffeic and p-coumaric acids. Acetic acid bacteria had minimal performance with respect to volatile acidity production in comparison to commercial vinegars. Acetic acid bacteria selection should therefore be reconsidered and the decrease of certain phenolic acids during acetous fermentation needs to be investigated.

Keywords: acetic acid bacteria, liquid chromatography, phenolics, saccharomyces cerevisiae, schizosaccharomyces pombe

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1844 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 212
1843 Optimization of Cacao Fermentation in Davao Philippines Using Sustainable Method

Authors: Ian Marc G. Cabugsa, Kim Ryan Won, Kareem Mamac, Manuel Dee, Merlita Garcia


An optimized cacao fermentation technique was developed for the cacao farmers of Davao City Philippines. Cacao samples with weights ranging from 150-250 kilograms were collected from various cacao farms in Davao City and Zamboanga City Philippines. Different fermentation techniques were used starting with design of the sweat box, prefermentation conditionings, number of days for fermentation and number of turns. As the beans are being fermented, its temperature was regularly monitored using a digital thermometer. The resultant cacao beans were assessed using physical and chemical means. For the physical assessment, the bean cut test, bean count tests, and sensory test were used. Quantification of theobromine, caffeine, and antioxidants in the form of equivalent quercetin was used for chemical assessment. Both the theobromine and caffeine were analyzed using HPLC method while the antioxidant was analyzed spectrometrically. To come up with the best fermentation procedure, the different assessment were given priority coefficients wherein the physical tests – taste test, cut, and bean count tests were given priority over the results of the chemical test. The result of the study was an optimized fermentation protocol that is readily adaptable and transferable to any cacao cooperatives or groups in Mindanao or even Philippines as a whole.

Keywords: cacao, fermentation, HPLC, optimization, Philippines

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