Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 58

Fermentation Related Abstracts

58 New Methodology for Monitoring Alcoholic Fermentation Processes Using Refractometry

Authors: Boukhiar Aissa, Halladj Fatima, Iguergaziz Nadia, 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: Alcohol, Fermentation, refractometry, residual sugar, brix, date, juice

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57 Device for Thermo-Magnetic Depolymerisation of Plant Biomass Prior to Methane Fermentation

Authors: Marcin Debowski, Marcin Zielinski, Mirosław Krzemieniewski


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

Keywords: biomass, Pre-treatment, Fermentation, depolymerisation

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56 Effect of Different Processing Methods on the Proximate, Functional, Sensory, and Nutritional Properties of Weaning Foods Formulated from Maize (Zea mays) and Soybean (Glycine max) Flour Blends

Authors: C. C. Okafor, C. O. Agu


Maize and soybean flours were produced using different methods of processing which include fermentation (FWF), roasting (RWF) and malting (MWF). Products from the different methods were mixed in the ratio 60:40 maize/soybean, respectively. These composites mixed with other ingredients such as sugar, vegetable oil, vanilla flavour and vitamin mix were analyzed for proximate composition, physical/functional, sensory and nutritional properties. The results for the protein content ranged between 6.25% and 16.65% with sample RWF having the highest value. Crude fibre values ranged from 3.72 to 10.0%, carbohydrate from 58.98% to 64.2%, ash from 1.27 to 2.45%. Physical and functional properties such as bulk density, wettability, gelation capacity have values between 0.74 and 0.76g/ml, 20.33 and 46.33 min and 0.73 to 0.93g/ml, respectively. On the sensory quality colour, flavour, taste, texture and general acceptability were determined. In terms of colour and flavour there was no significant difference (P < 0.05) while the values for taste ranged between 4.89 and 7.1 l, texture 5.50 to 8.38 and general acceptability 6.09 and 7.89. Nutritionally there is no significant difference (P < 0.05) between sample RWF and the control in all parameters considered. Samples FWF and MWF showed significantly (P < 0.5) lower values in all parameters determined. In the light of the above findings, roasting method is highly recommend in the production of weaning foods.

Keywords: Fermentation, Wettability, malting, ratio, roasting

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55 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: Fermentation, legume, condiment, tamarind seed

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54 Preliminary Study of Fermented Pickle of Tabah Bamboo Shoot: Gigantochloa nigrociliata (Buese) Kurz

Authors: Luh Putu T. Darmayanti, A. A. Duwipayana, Nyoman S. Antara, I. Nengah K. Putra


Tabah Bamboo (Gigantochloa nigrociliata (Buese) Kurz) is the indigenous bamboo species which grows in District of Pupuan, Tabanan at Province of Bali. Compared to the others, this shoot has low concentration of hydrocyanide acid (HCN). However, as found for almost of bamboo shoot, its seasonal availability, perishable in nature, and short-lived. This study aimed to gather information about total of lactic acid bacteria (LAB), pH, total acidity, HCN content, detection of LAB’s type involved during fermentation, and organic acids’ profiles of fermented pickles of Tabah bamboo shoot. The pickle was made by natural fermentation with 6 % salt concentration and fermentation conducted for 13 days. The result showed during the fermentation time, in the fourth day we found LAB’s number was highest as much as 72 x 107 CFU/ml and the lowest pH was 3.09. We also found decreasing in HCN from 37.8 ppm at the beginning to 20.52 ppm at the end of fermentation process. The total number of indigenous LAB isolated from the pickle are 48 strains we found 18 out of these had rod shape. For the preliminary study, all of the LAB with rod shape were detected by PCR as member of Lactobacillus spp., in which 17 strains detected as L. plantarum. The organic acids detected during the fermentation were lactic acid with the highest concentration was 0.0546 g/100 g and small amount of acetic acid.

Keywords: Fermentation, LAB, pickle, Tabah Bamboo shoot

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53 Dependence of the Electro-Stimulation of Saccharomyces cerevisiae by Pulsed Electric Field at the Yeast Growth Phase

Authors: Henri El Zakhem, Jessy Mattar, Mohamad Turk, Maurice Nonus, Nikolai Lebovka, Eugene Vorobiev


The effects of electro-stimulation of S. cerevisiae cells in colloidal suspension by Pulsed Electric Fields ‎‎(PEF) with electric field strength E = 20 – 2000 and effective PEF treatment time tPEF = 10^−5 – 1 s were ‎investigated. The applied experimental procedure includes variations in the preliminary fermentation time and ‎electro-stimulation by PEF-treatment. Plate counting was performed.‎ At relatively high electric fields (E ≥ 1000 and moderate PEF treatment time (tPEF > 100 µs), the ‎extraction of ionic components from yeast was observed by conductivity measurements, which can be related to ‎electroporation of cell membranes. Cell counting revealed a dependency of the colonies’ size on the time of ‎preliminary fermentation tf and the power consumption W, however no dependencies were noticeable by varying the initial yeast concentration in the treated suspensions.‎

Keywords: Biotechnology, Fermentation, Yeast, Electroporation, intensification

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52 Effect of Temperature on the Production of Fructose and Bioethanol from Date’s Syrup using S. cerevisiae ATCC 36859

Authors: M. A. Zeinelabdeen, A. E. Abasaeed, M. H. Gaily, A. K. Sulieman, M. D. Putra


The effect of temperature on the production of fructose and bioethanol from date syrup via selective fermentation by S. cerevisiae ATCC 36859 strain was studied. Various temperatures have been tested (27, 30 and 33 ᵒC). The fermentation experiments were conducted in a water shaker bath at the three temperatures under testing and 120 rpm. The results showed that a high fructose yield can be achieved at all temperatures under testing while the optimal is 27 ᵒC with 84% fructose yield. A high ethanol yield can be obtained for all temperatures under testing. However; the maximum biomass concentration and ethanol yield (86.22%) were obtained at 30 ᵒC.

Keywords: Fermentation, Ethanol, fructose, dates, S. cerevisiae

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51 Bioproduction of L(+)-Lactic Acid and Purification by Ion Exchange Mechanism

Authors: Zelal Polat, Şebnem Harsa, Semra Ülkü


Lactic acid exists in nature optically in two forms, L(+), D(-)-lactic acid, and has been used in food, leather, textile, pharmaceutical and cosmetic industries. Moreover, L(+)-lactic acid constitutes the raw material for the production of poly-L-lactic acid which is used in biomedical applications. Microbially produced lactic acid was aimed to be recovered from the fermentation media efficiently and economically. Among the various downstream operations, ion exchange chromatography is highly selective and yields a low cost product recovery within a short period of time. In this project, Lactobacillus casei NRRL B-441 was used for the production of L(+)-lactic acid from whey by fermentation at pH 5.5 and 37°C that took 12 hours. The product concentration was 50 g/l with 100% L(+)-lactic acid content. Next, the suitable resin was selected due to its high sorption capacity with rapid equilibrium behavior. Dowex marathon WBA, weakly basic anion exchanger in OH form reached the equilibrium in 15 minutes. The batch adsorption experiments were done approximately at pH 7.0 and 30°C and sampling was continued for 20 hours. Furthermore, the effect of temperature and pH was investigated and their influence was found to be unimportant. All the adsorption/desorption experiments were applied to both model lactic acid and biomass free fermentation broth. The ion exchange equilibria of lactic acid and L(+)-lactic acid in fermentation broth on Dowex marathon WBA was explained by Langmuir isotherm. The maximum exchange capacity (qm) for model lactic acid was 0.25 g La/g wet resin and for fermentation broth 0.04 g La/g wet resin. The equilibrium loading and exchange efficiency of L(+)-lactic acid in fermentation broth were reduced as a result of competition by other ionic species. The competing ions inhibit the binding of L(+)-lactic acid to the free sites of ion exchanger. Moreover, column operations were applied to recover adsorbed lactic acid from the ion exchanger. 2.0 M HCl was the suitable eluting agent to recover the bound L(+)-lactic acid with a flowrate of 1 ml/min at ambient temperature. About 95% of bound L(+)-lactic acid was recovered from Dowex marathon WBA. The equilibrium was reached within 15 minutes. The aim of this project was to investigate the purification of L(+)-lactic acid with ion exchange method from fermentation broth. The additional goals were to investigate the end product purity, to obtain new data on the adsorption/desorption behaviours of lactic acid and applicability of the system in industrial usage.

Keywords: Purification, Fermentation, ion exchange, whey, lactic acid

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50 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: Fermentation, microalgal biomass, enzymatic saccharification, biobutanol

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49 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: Fermentation, bio-hydrogen, experimental bioreactor, crude glycerol

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

Authors: Kamla Malik


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

Keywords: Fermentation, Ethanol, fruit wastes, paddy straw

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47 The Nutritive Value of Fermented Sago Pith (Metroxylon sago Rottb) Enriched with Micro Nutrients for Poultry Feed

Authors: Wizna, Helmi Muis, Hafil Abbas


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

Keywords: Fermentation, sulfur, urea, sago pith, Bacillus amyloliquefaciens

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46 Optimization of Media for Enhanced Fermentative Production of Mycophenolic Acid by Penicillium brevicompactum

Authors: Shraddha Digole, Swarali Hingse, Uday Annapure


Mycophenolic acid (MPA) is an immunosuppressant; produced by Penicillium Sp. Box-Behnken statistical experimental design was employed to optimize the condition of Penicillium brevicompactum NRRL 2011 for mycophenolic acid (MPA) production. Initially optimization of various physicochemical parameters and media components was carried out using one factor at a time approach and significant factors were screened by Taguchi L-16 orthogonal array design. Taguchi design indicated that glucose, KH2PO4 and MgSO4 had significant effect on MPA production. These variables were selected for further optimization studies using Box-Behnken design. Optimised fermentation condition, glucose (60 g/L), glycine (28 g/L), L-leucine (1.5g/L), KH2PO4 (3g/L), MgSO4.7H2O (1.5g/L), increased the production of MPA from 170 mg/L to 1032.54 mg/L. Analysis of variance (ANOVA) showed a high value of coefficient of determination R2 (0.9965), indicating a good agreement between experimental and predicted values and proves validity of the statistical model.

Keywords: Fermentation, Box-Behnken design, mycophenolic acid, Penicillium brevicompactum

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

Authors: S. James, Ocheme B. Ocheme, Omobolanle O. Oloyede, 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, functional properties, moringa, flour

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44 Integrated Process Modelling of a Thermophilic Biogas Plant

Authors: Jeremiah L. Chukwuneke, Obiora E. Anisiji, Chinonso H. Achebe, Paul C. Okolie


This work developed a mathematical model of a biogas plant from a mechanistic point of view, for urban area clean energy requirement. It aimed at integrating thermodynamics; which deals with the direction in which a process occurs and Biochemical kinetics; which gives the understanding of the rates of biochemical reaction. The mathematical formulation of the proposed gas plant follows the fundamental principles of thermodynamics, and further analysis were accomplished to develop an algorithm for evaluating the plant performance preferably in terms of daily production capacity. In addition, the capacity of the plant is equally estimated for a given cycle of operation and presented in time histories. A nominal 1500m3 biogas plant was studied characteristically and its performance efficiency evaluated. It was observed that the rate of biogas production is essentially a function of enthalpy ratio, the reactor temperature, pH, substrate concentration, rate of degradation of the biomass, and the accumulation of matter in the system due to bacteria growth. The results of this study conform to a very large extent with reported empirical data of some existing plant and further model validations were conducted in line with classical records found in literature.

Keywords: Energy, Fermentation, temperature, biogas production, Anaerobic Digestion, bio-reactor, biogas plant, rate of production, therm

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43 Effects of Varying Fermentation Periods on the Chemical Composition of African Yam Bean (Sphenostylis stenocarpa) and Acha (Digitaria exilis) Flour Blends and Sensory Properties of Their Products

Authors: P. N. Okeke, J. N. Chikwendu


The study evaluated the effects of varying fermentation periods on the nutrients and anti-nutrients composition of African yam bean (Sphenostylis stenocarpa) and acha (Digitaria exilis) flour blends and sensory properties of their products. The African yam bean seeds and acha grains were fermented for 24 hrs, 48 and 72 hrs, dried (sun drying) and milled into fine flour. The fermented flours were used in a ratio of 70:30 (Protein basis) to formulate composite flour for meat pie and biscuits production. Both the fermented and unfermented flours and products were analyzed for chemical composition using the standard method. The data were statistically analyzed using SPSS version 15 to determine the mean and standard deviation. The 24, 48, and 72 hrs fermentation periods increased protein (22.81, 26.15 and 24.00% respectively). The carbohydrate, ash and moisture contents of the flours were also increased as a result of fermentation (68.01-76.83, 2.26-4.88, and 8.36-13.00% respectively). The 48 hrs fermented flour blends had the highest increase in ash relative to the control (4.88%). Fermentation increased zinc, iron, magnesium and phosphorus content of the flours. Treatment drastically reduced the anti-nutrient (oxalate, saponin, tannin, phytate, and hemagglutinin) levels of the flours. Both meat pie and biscuits had increased protein relative to the control (27.36-34.28% and 23.66-25.09%). However, the protein content of the meat pie increased more than that of the biscuits. Zinc, Iron, Magnesium and phosphorus levels increased in both meat pie and biscuits. Organoleptic attributes of the products (meat pie and biscuits) were slightly lower than the control except those of the 72 hrs fermented flours.

Keywords: Fermentation, acha, African yam bean, biscuits, meat-pie

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42 Hyper-Production of Lysine through Fermentation and Its Biological Evaluation on Broiler Chicks

Authors: Shagufta Gulraiz, Abu Saeed Hashmi, Muhammad Mohsin Javed


Lysine required for poultry feed is imported in Pakistan to fulfil the desired dietary needs. Present study was designed to produce maximum lysine by utilizing cheap sources to save the foreign exchange. To achieve the goal of lysine production through fermentation, large scale production of lysine was carried out in 7.5 L stirred glass vessel fermenter with wild and mutant Brevibacterium flavum (B. flavum) using all pre-optimized conditions. The identification of produced lysine was carried out by TLC and amino acid analyzer. Toxicity evaluation of produced lysine was performed before feeding to broiler chicks. During biological trial concentrated fermented broth having 8% lysine was used in poultry rations as a source of Lysine for test birds. Fermenter scale studies showed that the maximum lysine (20.8 g/L) was produced at 250 rpm, 1.5 vvm aeration, 6.0% inoculum under controlled pH conditions after 56 h of fermentation with wild culture but mutant (BFENU2) gave maximum yield of lysine 36.3 g/L under optimized condition after 48 h. Amino acid profiling showed 1.826% Lysine in fermented broth by wild B. flavum and 2.644% by mutant strain (BFENU2). Toxicity evaluation report showed that the produced lysine is safe for consumption by broilers. Biological evaluation results showed that produced lysine was equally good as commercial lysine in terms of weight gain, feed intake and feed conversion ratio. A cheap and practical bioprocess of Lysine production was concluded, that can be exploited commercially in Pakistan to save foreign exchange.

Keywords: Fermentation, broiler chicks, lysine, biological evaluation

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41 Process Optimization and Microbial Quality of Provitamin A-Biofortified Amahewu, a Non-Alcoholic Maize Based Beverage

Authors: Oluwatosin A. Ijabadeniyi, Temitope D. Awobusuyi, Eric O. Amonsou, Muthulisi Siwela


Provitamin A-biofortified maize has been developed to alleviate Vitamin A deficiency; a major public health problem in developing countries. Amahewu, a non-alcoholic fermented maize based beverage is produced using white maize, which is deficient in Vitamin A. In this study, the suitable processing conditions for the production of amahewu using provitamin A-biofortified maize and the microbial quality of the processed products were evaluated. Provitamin A-biofortified amahewu was produced with reference to traditional processing method. Processing variables were Inoculum types (Malted provitamin A maize, Wheat bran, and lactobacillus mixed starter culture with either malted provitamin A or wheat bran) and concentration (0.5 %, 1 % and 2 %). A total of four provitamin A-biofortified amahewu products after fermentation were subjected to different storage conditions: 4ᴼC, 25ᴼC and 37ᴼC. pH and TTA were monitored throughout the storage period. Sample of provitamin A-biofortified amahewu were plated and observed every day for 5 days to assess the presence of Aerobic and Anaerobic spore formers, E.coli, Lactobacillus and Mould. The addition of starter culture substantially reduced the fermentation time (6 hour, pH 3.3) compared to those with no addition of starter culture (24 hour pH 3.5). It was observed that Lactobacillus were present from day 0 for all the storage temperatures. The presence of aerobic spore former and mould were observed on day 3. E.coli and Anaerobic spore formers were not present throughout the storage period. These microbial growth were minimal at 4ᴼC while 25ᴼC had higher counts of growth with 37ᴼC having the highest colony count. Throughout the storage period, pH of provitamin A-biofortified amahewu was stable. Provitamin A-biofortified amahewu stored under refrigerated condition (4ᴼC) had better storability compared to 25ᴼC and 37ᴼC. The production and microbial quality of provitamin A-biofortified amahewu might be important in combating Vitamin A Deficiency.

Keywords: Fermentation, biofortification, maize, vitamin A deficiency

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40 Fermented Unripe Plantain (Musa paradisiacal) Peel Meal as a Replacement for Maize in the Diet of Nile Tilapia (Oreochromis niloticus) Fingerlings

Authors: S. O. Obasa, N. A. Bamidele, O. E. Babalola, I. O. Taiwo, I. Abdulraheem, O. C. Odebiyi, A. A. Adeoye, O. V. Uzamere


A feeding trial was conducted to investigate the effect of fermented unripe plantain peel meal (FUP) on growth performance, nutrients digestibility and economic indices of production of Nile tilapia, Oreochromis niloticus fingerlings. Fingerlings (150) of Nile tilapia (1.70±0.1g) were stocked at 10 per plastic tank. Five iso-nitrogenous diets containing 40% crude protein in which maize meal was replaced by fermented unripe plantain peel meal at 0% (FUP0), 25% (FUP25), 50% (FUP50), 75% (FUP75) and 100% (FUP100) were formulated and prepared. The fingerlings were fed at 5% body weight per day for 56 days. There was no significant difference (p > 0.05) in all the growth parameters among the treatments. Feed conversion ratio of 1.35 in fish fed diet FUP25 was not significantly different (P > 0.05) from 1.42 of fish fed diet FUP0. Apparent protein digestibility of 86.94% in fish fed diet FUP100 was significantly higher (p < 0.05) than 70.37% in fish fed diet FUP0 while apparent carbohydrate of 88.34% in fish fed diet FUP0 was significantly different (p < 0.05) from 70.29% of FUP100. Red blood cell (4.30 ml/mm3) of fish fed diet FUP100 was not significantly different from 4.13 ml/mm3 of fish fed diet FUP50. The highest percentage profit of 88.85% in fish fed diet FUP100 was significantly higher than 66.68% in fish fed diet FUP0 while the profit index of 1.89 in fish fed diet FUP100 was significantly different from 1.67 in fish fed diet FUP0. Therefore, fermented unripe plantain peel meal can completely replace maize in the diet of O. niloticus fingerlings.

Keywords: Fermentation, tilapia, fish diets, plantain peel

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39 Bacteriological Quality of Commercially Prepared Fermented Ogi (AKAMU) Sold in Some Parts of South Eastern Nigeria

Authors: Alloysius C. Ogodo, Ositadinma C. Ugbogu, Uzochukwu G. Ekeleme


Food poisoning and infection by bacteria are of public health significance to both developing and developed countries. Samples of ogi (akamu) prepared from white and yellow variety of maize sold in Uturu and Okigwe were analyzed together with the laboratory prepared ogi for microbial quality using the standard microbiological methods. The analyses showed that both white and yellow variety had total bacterial counts (cfu/g) of 4.0 ×107 and 3.9 x 107 for the laboratory prepared ogi while the commercial ogi had 5.2 x 107 and 4.9 x107, 4.9 x107 and 4.5 x107, 5.4 x107 and 5.0 x107 for Eke-Okigwe, Up-gate and Nkwo-Achara market respectively. The Staphylococcal counts ranged from 2.0 x 102 to 5.0 x102 and 1.0 x 102 to 4.0 x102 for the white and yellow variety from the different markets while Staphylococcal growth was not recorded on the laboratory prepared ogi. The laboratory prepared ogi had no Coliform growth while the commercially prepared ogi had counts of 0.5 x103 to 1.6 x 103 for white variety and 0.3 x 103 to 1.1 x103 for yellow variety respectively. The Lactic acid bacterial count of 3.5x106 and 3.0x106 was recorded for the laboratory ogi while the commercially prepared ogi ranged from 3.2x106 to 4.2x106 (white variety) and 3.0 x106 to 3.9 x106 (yellow). The presence of bacteria isolates from the commercial and laboratory fermented ogi showed that Lactobacillus sp, Leuconostoc sp and Citrobacter sp were present in all the samples, Micrococcus sp and Klebsiella sp were isolated from Eke-Okigwe and ABSU-up-gate markets varieties respectively, E. coli and Staphylococcus sp were present in Eke-Okigwe and Nkwo-Achara markets while Salmonella sp were isolated from the three markets. Hence, there are chances of contracting food borne diseases from commercially prepared ogi. Therefore, there is the need for sanitary measures in the production of fermented cereals so as to minimize the rate of food borne pathogens during processing and storage.

Keywords: Lactic Acid Bacteria, Fermentation, maize, ogi, bacterial quality

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38 Fermentable Bio-Ethanol Using Bakers and Palmwine Yeasts: Indices of Bioavailability of Carbohydrate and Sugar from Fungal Treated Rice Husk

Authors: Ezeonu, Chukwuma Stephen, Onwurah, Ikechukwu Noel Emmanuel


Pure strains of Aspergillus fumigatus (AF), aspergillus niger (AN), aspergillus oryzae (AO), trichophyton mentagrophyte (TM), trichophyton rubrum (TR) and Trichophyton soudanense (TS) were isolated from decomposing rice husk. Freshly processed rice husk in Mandle’s medium were heat pre-treated using an autoclave at 121oC for 20 minutes. The isolated fungi as monoculture and di-culture combinations were inoculated into each of the pre-treated rice husk with the exception of two controls. Seven days hydrolysis was followed by estimation of carbohydrate, reducing sugar and non-reducing sugar. Fungal treated rice husks were left to ferment for 7 days with introduction of both baker’s and palm wine yeast. The result obtained in the work gave the highest carbohydrate (20.53 ± 2.73 %) from rice husks treated with TS + TR di-culture. The highest soluble reducing sugar (2.66 ± 0.14 %) was obtained from rice husk treated with TM. The highest soluble nonreducing sugar (18.08 ± 2.61 %) was from AF. The introduction of yeasts from palm wine gave the highest bio-ethanol (12.82 ± 0.39 %) from AO. The highest bio-ethanol (6.60 ± 0.10 %) from baker's yeast fermentation was in AO + TS treated rice husk. There was increased availability of sugar and moderate yield of bio-ethanol, especially from palm wine yeast.

Keywords: Fermentation, Carbohydrate, Fungi, Ethanol, rice husk, reducing sugar, non-reducing sugar

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37 Preservation of Coconut Toddy Sediments as a Leavening Agent for Bakery Products

Authors: B. R. Madushan, S. B. Navaratne, I. Wickramasinge


Toddy sediment (TS) was cultured in a PDA medium to determine initial yeast load, and also it was undergone sun, shade, solar, dehumidified cold air (DCA) and hot air oven (at 400, 500 and 60oC) drying with a view to preserve viability of yeast. Thereafter, this study was conducted according to two factor factorial design in order to determine best preservation method. Therein the dried TS from the best drying method was taken and divided into two portions. One portion was mixed with 3: 7 ratio of TS: rice flour and the mixture was divided in to two again. While one portion was kept under in house condition the other was in a refrigerator. Same procedure was followed to the rest portion of TS too but it was at the same ratio of corn flour. All treatments were vacuum packed in triple laminate pouches and the best preservation method was determined in terms of leavening index (LI). The TS obtained from the best preservation method was used to make foods (bread and hopper) and organoleptic properties of it were evaluated against same of ordinary foods using sensory panel with a five point hedonic scale. Results revealed that yeast load or fresh TS was 58×106 CFU/g. The best drying method in preserving viability of yeast was DCA because LI of this treatment (96%) is higher than that of other three treatments. Organoleptic properties of foods prepared from best preservation method are as same as ordinary foods according to Duo trio test.

Keywords: Fermentation, Yeast, biological leavening agent, coconut toddy

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36 Effects of Rumen Protozoa and Nitrate on Fermentation and Methane Production

Authors: L. Li, S. H. Nguyen, R. S. Hegarty


Two experiments were conducted assessing the effects of presence or absence of rumen protozoa and dietary nitrate addition on rumen fermentation characteristics and methane production in Brahman heifers. The first experiment assessed changes in rumen fermentation pattern and in-vitro methane production post-refaunation and the second experiment investigated whether addition of nitrate to the incubation would give rise to methane mitigation additional to that contributed by defaunation. Ten Brahman heifers were progressively adapted to a diet containing coconut oil distillate 4.5% (COD) for 18 d and then all heifers were defaunated using sodium 1-(2-sulfonatooxyethoxy) dodecane (Empicol). After 15 d, the heifers were given a second dose of Empicol. Fifteen days after the second dosing, all heifers were allocated to defaunated or refaunated groups by stratified randomisation. On d 48, an oral dose of rumen fluid collected from unrelated faunated cattle was used to inoculate 5 heifers and form a refaunated group so that the effects of re-establishment of protozoa on fermentation characteristics could be investigated. Samples of rumen fluid collected from each animal using oesophageal intubation before feeding on d 48, 55, 62 and 69 were incubated for 23h in-vitro (experiment 1). On day 82, 2% of NO3 (as NaNO3) was included in in-vitro incubations (experiment 2) to test for additivity of NO3 and absence of protozoa effects on fermentation and methane production. It was concluded that increasing protozoal numbers were associated with increased methane production, with methane production rate significantly higher from refaunated heifers than from defaunated heifers 7, 14 and 21 d after refaunation. Concentration and proportions of major VFA, however, were not affected by protozoal treatments. There is scope for further reducing methane output through combining defaunation and dietary nitrate as the addition of nitrate in the defaunated heifers resulted in 86% reduction in methane production in-vitro.

Keywords: Fermentation, Methane Production, nitrate, defaunation

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35 Optimizing Fermented Paper Production Using Spyrogira sp. Interpolating with Banana Pulp

Authors: Hadiatullah, T. S. D. Desak Ketut, A. A. Ayu, A. N. Isna, D. P. Ririn


Spirogyra sp. is genus of microalgae which has a high carbohydrate content that used as a best medium for bacterial fermentation to produce cellulose. This study objective to determine the effect of pulp banana in the fermented paper production process using Spirogyra sp. and characterizing of the paper product. The method includes the production of bacterial cellulose, assay of the effect fermented paper interpolating with banana pulp using Spirogyra sp., and the assay of paper characteristics include gram-mage paper, water assay absorption, thickness, power assay of tensile resistance, assay of tear resistance, density, and organoleptic assay. Experiments were carried out with completely randomized design with a variation of the concentration of sewage treatment in the fermented paper production interpolating banana pulp using Spirogyra sp. Each parameter data to be analyzed by Anova variance that continued by real difference test with an error rate of 5% using the SPSS. Nata production results indicate that different carbon sources (glucose and sugar) did not show any significant differences from cellulose parameters assay. Significantly different results only indicated for the control treatment. Although not significantly different from the addition of a carbon source, sugar showed higher potency to produce high cellulose. Based on characteristic assay of the fermented paper showed that the paper gram-mage indicated that the control treatment without interpolation of a carbon source and a banana pulp have better result than banana pulp interpolation. Results of control gram-mage is 260 gsm that show optimized by cardboard. While on paper gram-mage produced with the banana pulp interpolation is about 120-200 gsm that show optimized by magazine paper and art paper. Based on the density, weight, water absorption assays, and organoleptic assay of paper showing the highest results in the treatment of pulp banana interpolation with sugar source as carbon is 14.28 g/m2, 0.02 g and 0.041 g/cm2.minutes. The conclusion found that paper with nata material interpolating with sugar and banana pulp has the potential formulation to produce super-quality paper.

Keywords: Fermentation, Cellulose, Spirogyra sp, paper, grammage

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34 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: Fermentation, Colour, cocoa beans, shallow box

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33 Comparative Study on Sensory Profiles of Liquor from Different Dried Cocoa Beans

Authors: Khairul Bariah Sulaiman, Tajul Aris Yang


Malaysian dried cocoa beans have been reported to have low quality flavour and are often sold at discounted prices. Various efforts have been made to improve the Malaysian beans quality. Among these efforts is introduction of the shallow box fermentation technique and pulp preconditioned through pods storage. However, after nearly four decades of the effort was done, Malaysian cocoa farmers still received lower prices for their beans. So, this study was carried out in order to assess the flavour quality of dried cocoa beans produced by shallow box fermentation techniques, combination of shallow box fermentation with pods storage and compared to dried cocoa beans obtained from Ghana. A total of eight samples of dried cocoa was used in this study, which one of the samples was Ghanaian beans (coded with no.8), while the rest were Malaysian cocoa beans with different post-harvest processing (coded with no. 1, 2, 3, 4, 5, 6 and 7). Cocoa liquor was prepared from all samples in the prescribed techniques and sensory evaluation was carried out using Quantitative Descriptive Analysis (QDA) Method with 0-10 scale by Malaysian Cocoa Board trained panelist. Sensory evaluation showed that cocoa attributes for all cocoa liquors ranging from 3.5 to 5.3, whereas bitterness was ranging from 3.4 to 4.6 and astringent attribute ranging from 3.9 to 5.5, respectively. Meanwhile, all cocoa liquors were having acid or sourness attribute ranging from 1.6 to 3.6, respectively. In general cocoa liquor prepared from sample coded no 4 has almost similar flavour profile and no significantly different at p < 0.05 with Ghana, in term of most flavour attributes as compared to the other six samples.

Keywords: Fermentation, cocoa beans, shallow box, flavour, pods storage

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32 Production of Human BMP-7 with Recombinant E. coli and B. subtilis

Authors: Jong Il Rhee


The polypeptide representing the mature part of human BMP-7 was cloned and efficiently expressed in Escherichia coli and Bacillus subtilis, which had a clear band for hBMP-7, a homodimeric protein with an apparent molecular weight of 15.4 kDa. Recombinant E.coli produced 111 pg hBMP-7/mg of protein hBMP-7 through IPTG induction. Recombinant B. subtilis also produced 350 pg hBMP-7/ml of culture medium. The hBMP-7 was purified in 2 steps using an FPLC system with an ion exchange column and a gel filtration column. The hBMP-7 produced in this work also stimulated the alkaline phosphatase (ALP) activity in a dose-dependent manner, i.e. 2.5- and 8.9-fold at 100 and 300 ng hBMP-7/ml, respectively, and showed intact biological activity.

Keywords: Fermentation, E. coli, B. subtilis, hBMP-7

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31 Alternative Energy and Carbon Source for Biosurfactant Production

Authors: Akram Abi, Mohammad Hossein Sarrafzadeh


Because of their several advantages over chemical surfactants, biosurfactants have given rise to a growing interest in the past decades. Advantages such as lower toxicity, higher biodegradability, higher selectivity and applicable at extreme temperature and pH which enables them to be used in a variety of applications such as: enhanced oil recovery, environmental and pharmaceutical applications, etc. Bacillus subtilis produces a cyclic lipopeptide, called surfactin, which is one of the most powerful biosurfactants with ability to decrease surface tension of water from 72 mN/m to 27 mN/m. In addition to its biosurfactant character, surfactin exhibits interesting biological activities such as: inhibition of fibrin clot formation, lyses of erythrocytes and several bacterial spheroplasts, antiviral, anti-tumoral and antibacterial properties. Surfactin is an antibiotic substance and has been shown recently to possess anti-HIV activity. However, application of biosurfactants is limited by their high production cost. The cost can be reduced by optimizing biosurfactant production using cheap feed stock. Utilization of inexpensive substrates and unconventional carbon sources like urban or agro-industrial wastes is a promising strategy to decrease the production cost of biosurfactants. With suitable engineering optimization and microbiological modifications, these wastes can be used as substrates for large-scale production of biosurfactants. As an effort to fulfill this purpose, in this work we have tried to utilize olive oil as second carbon source and also yeast extract as second nitrogen source to investigate the effect on both biomass and biosurfactant production improvement in Bacillus subtilis cultures. Since the turbidity of the culture was affected by presence of the oil, optical density was compromised and no longer could be used as an index of growth and biomass concentration. Therefore, cell Dry Weight measurements with applying necessary tactics for removing oil drops to prevent interference with biomass weight were carried out to monitor biomass concentration during the growth of the bacterium. The surface tension and critical micelle dilutions (CMD-1, CMD-2) were considered as an indirect measurement of biosurfactant production. Distinctive and promising results were obtained in the cultures containing olive oil compared to cultures without it: more than two fold increase in biomass production (from 2 g/l to 5 g/l) and considerable reduction in surface tension, down to 40 mN/m at surprisingly early hours of culture time (only 5hr after inoculation). This early onset of biosurfactant production in this culture is specially interesting when compared to the conventional cultures at which this reduction in surface tension is not obtained until 30 hour of culture time. Reducing the production time is a very prominent result to be considered for large scale process development. Furthermore, these results can be used to develop strategies for utilization of agro-industrial wastes (such as olive oil mill residue, molasses, etc.) as cheap and easily accessible feed stocks to decrease the high costs of biosurfactant production.

Keywords: Fermentation, biosurfactant, Bacillus subtilis, surfactin, agro-industrial waste, second carbon and nitrogen source

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

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


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

Keywords: Fermentation, bacterial cellulose, aeration, gluconacetobacter xylinus

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29 Enhanced Anti-Obesity Effect of Soybean by Fermentation with Lactobacillus plantarum P1201 in 3T3-L1 Adipocyte

Authors: Chengliang Xie, Jinhyun Ryu, Hyun Joon Kim, Gyeong Jae Cho, Wan Sung Choi, Sang Soo Kang, Kye Man Cho, Dong Hoon Lee


Obesity has become a global health problem and a source of major metabolic diseases like type-2 diabetes, hypertension, heart disease, nonalcoholic fatty liver and cancer. Synthetic anti-obesity drugs are effective but very costly and with undesirable side effects, so natural products such as soybean are needed as an alternative for obesity treatment. Lactobacillus Plantarum P1201is a probiotic bacterial strain reported to produce conjugated linoleic acid (CLA) and increase the ratio of aglycone-isoflavone of soybean, both of which have anti-obesity effect. In this study, the anti-obesity effect of the fermented soybean extract with P1201 (FSE) will be evaluated compared with that of the soybean extract (SE) by 3T3-L1 cells as an in vitro model of adipogenesis. 3T3-L1 cells were treated with SE and FSE during the nine days of the differentiation, lipid accumulation was evaluated by oil-red staining and triglyceride content and the mRNA expression level of adipogenic or lipogenic genes were analyzed by RT-PCR and qPCR. The results showed that formation of lipid droplets in differentiated 3T3-L1 cells was inhibited and triglyceride content was reduced by 23.1% after treated with 1000 μg/mL of FSE compared with control. For SE-treated groups, no delipidating effect was observed. The effect of FSE on adipogenesis inhibition can be attributed to the down-regulation of mRNA expressionof CCAAT/enhancer binding protein (C/EBP-α), lipoprotein lipase (LPL), adiponectin, adipocyte fatty acid-binding protein (aP2), fatty acid synthesis (FAS) and CoA carboxylase (ACC). Our results demonstrated that the anti-obesity effect of soybean can be improved by fermentation with P1201, and P1201can be used as a potential probiotic bacterial strain to produce natural anti-obesity food.

Keywords: Obesity, Fermentation, soybean, Lactobacillus plantarum P1201

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