Search results for: thermophilic fermentation

Authors: Sachin B. Patil, Narendra M. Kanhe

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A typical cane molasses based distillery generates 15 L of waste water per liter of alcohol production. Distillery waste with COD of over 1,00,000 mg/l and BOD of over 30,000 mg/l ranks high amongst the pollutants produced by industries both in magnitude and strength. Treatment and safe disposal of this waste is a challenging task since long. The high strength of waste water renders aerobic treatment very expensive and physico-chemical processes have met with little success. Thermophilic anaerobic treatment of distillery waste may provide high degree of treatment and better recovery of biogas. It may prove more feasible in most part of tropical country like India, where temperature is suitable for thermophilic micro-organisms. Researchers have reviled that, at thermophilic conditions due to increased destruction rate of organic matter and pathogens, higher digestion rate can be achieved. Literature review reveals that the variety of anaerobic reactors including anaerobic lagoon, conventional digester, anaerobic filter, two staged fixed film reactors, sludge bed and granular bed reactors have been studied, but little attempts have been made to evaluate the usefulness of thermophilic anaerobic treatment for treating distillery waste. The present study has been carried out, to study feasibility of thermophilic anaerobic digestion to facilitate the design of full scale reactor. A pilot scale anaerobic fixed film fixed bed reactor (AFFFB) of capacity 25m3 was designed, fabricated, installed and commissioned for thermophilic (55-65°C) anaerobic digestion at a constant pH of 6.5-7.5, because these temperature and pH ranges are considered to be optimum for biogas recovery from distillery wastewater. In these conditions, working of the reactor was studied, for different hydraulic retention times (HRT) (0.25days to 12days) and variable organic loading rates (361.46 to 7.96 Kg COD/m3d). The parameters such as flow rate and temperature, various chemical parameters such as pH, chemical oxygen demands (COD), biogas quantity, and biogas composition were regularly monitored. It was observed that, with the increase in OLR, the biogas production was increased, but the specific biogas yield decreased. Similarly, with the increase in HRT, the biogas production got decrease, but the specific biogas yield was increased. This may also be due to the predominant activity of acid producers to methane producers at the higher substrate loading rates. From the present investigation, it can be concluded that for thermophilic conditions the highest COD removal percentage was obtained at an HRT of 08 days, thereafter it tends to decrease from 8 to 12 days HRT. There is a little difference between COD removal efficiency of 8 days HRT (74.03%) and 5 day HRT (78.06%), therefore it would not be feasible to increase the reactor size by 1.5 times for mere 4 percent more efficiency. Hence, 5 days HRT is considered to be optimum, at which the biogas yield was 98 m3/day and specific biogas yield was 0.385 CH4 m3/Kg CODr.

Keywords: spent wash, anaerobic digestion, biomass, biogas

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

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

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

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

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

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

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

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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

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

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

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

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

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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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Authors: Farhan Saeed

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

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

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

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

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

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

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

Keywords: aeration, bacterial cellulose, fermentation, gluconacetobacter xylinus

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

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

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

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

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

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

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Authors: Kamla Malik

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

Keywords: ethanol, fermentation, fruit wastes, paddy straw

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Authors: Jožef Ritonja

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The biochemical industry is one of the most important modern industries. Biochemical reactors are crucial devices of the biochemical industry. The essential bioprocess carried out in bioreactors is the fermentation process. A thorough insight into the fermentation process and the knowledge how to control it are essential for effective use of bioreactors to produce high quality and quantitatively enough products. The development of the control system starts with the determination of a mathematical model that describes the steady state and dynamic properties of the controlled plant satisfactorily, and is suitable for the development of the control system. The paper analyses the fermentation process in bioreactors thoroughly, using existing mathematical models. Most existing mathematical models do not allow the design of a control system for controlling the fermentation process in batch bioreactors. Due to this, a mathematical model was developed and presented that allows the development of a control system for batch bioreactors. Based on the developed mathematical model, a control system was designed to ensure optimal response of the biochemical quantities in the fermentation process. Due to the time-varying and non-linear nature of the controlled plant, the conventional control system with a proportional-integral-differential controller with constant parameters does not provide the desired transient response. The improved adaptive control system was proposed to improve the dynamics of the fermentation. The use of the adaptive control is suggested because the parameters’ variations of the fermentation process are very slow. The developed control system was tested to produce dairy products in the laboratory bioreactor. A carbon dioxide concentration was chosen as the controlled variable. The carbon dioxide concentration correlates well with the other, for the quality of the fermentation process in significant quantities. The level of the carbon dioxide concentration gives important information about the fermentation process. The obtained results showed that the designed control system provides minimum error between reference and actual values of carbon dioxide concentration during a transient response and in a steady state. The recommended control system makes reference signal tracking much more efficient than the currently used conventional control systems which are based on linear control theory. The proposed control system represents a very effective solution for the improvement of the milk fermentation process.

Keywords: biochemical reactor, fermentation process, modelling, adaptive control

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

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

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

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

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

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

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

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

Keywords: tempe, anticancer, antioxidant, phenolic compounds

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

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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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Authors: L. Kutateldze, T. Urushadze, R. Khvedelidze, N. Zakariashvili, I. Khokhashvili, T. Sadunishvili

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Microbial cellulase/xylanase have shown their potential application in various industries including pulp and paper, textile, laundry, biofuel production, food and feed industry, brewing, and agriculture. Extremophilic micromycetes and their enzymes that are resistant to critical values of temperature and pH, and retaining enzyme activity for a long time are of great industrial interest. Among strains of microscopic fungi from the collection of S. Durmishidze Institute of Biochemistry and Biotechnology, strains isolated from different ecological niches of Southern Caucasus-active producers of cellulase/xylanase have been selected by means of screening under deep cultivation conditions. Extremophilic micromycetes and their enzymes that are resistant to critical values of temperature and pH, and retaining enzyme activity for a long time are of great industrial interest. Among strains of microscopic fungi from the collection of S. Durmishidze Institute of Biochemistry and Biotechnology, strains isolated from different ecological niches of Southern Caucasus-active producers of cellulase/xylanase have been selected by means of screening under deep cultivation conditions. Representatives of the genera Aspergillus, Penicillium and Trichoderma are outstanding by relatively high activities of these enzymes. Among the producers were revealed thermophilic strains, representatives of the genus Aspergillus-Aspergillus terreus, Aspergillus versicolor, Aspergillus wentii, also strains of Sporotrichum pulverulentum and Chaetomium thermophile. As a result of optimization of cultivation media and conditions, activities of enzymes produced by the strains have been increased by 4 -189 %. Two strains, active producers of cellulase/xylanase – Penicillium canescence E2 (mesophile) and Aspergillus versicolor Z17 (thermophile) were chosen for further studies. Cellulase/xylanase enzyme preparations from two different genera of microscopic fungi Penicillium canescence E2 and Aspergillus versicolor Z 17 were obtained with activities 220 U/g /1200 U/g and 125 U/g /940 U/g, correspondingly. Main technical characteristics were as follows: the highest enzyme activities were obtained for mesophilic strain Penicillium canescence E2 at 45-500C, while almost the same enzyme activities were fixed for the thermophilic strain Aspergillus versicolor Z 17 at temperature 60-65°C, exceeding the temperature optimum of the mesophile by 150C. Optimum pH of action of the studied cellulase/xylanases from mesophileic and thermophilic strains were similar and equaled to 4.5-5.0 It has been shown that cellulase/xylanase technical preparations from selected strains of Penicillium canescence E2 and Aspergillus versicolor Z17 hydrolyzed cellulose of untreated wheat straw to reducible sugars by 46-52%, and to glucose by 22-27%. However the thermophilic enzyme preparations from the thermophilic A.versicolor strains conducted the process at 600C higher by 100C as compared to mesophlic analogue. Rate of hydrolyses of the pretreated substrate by the same enzyme preparations to reducible sugars and glucose conducted at optimum for their action 60 and 500C was 52-61% and 29-33%, correspondingly. Thus, maximum yield of glucose and reducible sugars form untreated and pretreated wheat straw was achieved at higher temperature (600C) by enzyme preparations from thermophilic strain, which gives advantage for their industrial application.

Keywords: cellulase/xylanase, cellulose hydrolysis, microscopic fungi, thermophilic strain

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Authors: Arezoo Ghadi, Adonis Pishdadian, Ehsan Zahedi, Vahideh Rashedi, Mozhgan Mohammadi

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The presence of antibiotics in milk is one of the problems of dairy production units, especially yogurt and cheese, which leads to a decrease in lactic coagulation. Here, to assess the incubation conditions for the fermentation of milk containing antibiotics, concentrations of 50, 75, 100, and 200 ppb of tetracycline were added to each liter of milk. Inoculation process with starter culture performed at three temperatures of 35°C, 45°C, and 50°C. Afterward, pH, acidity, oxidation-reduction potential, and lactic coagulation of yogurt were evaluated. The results showed the existence of antibiotics in milk affects the quality and physicochemical properties of yogurt. However, antibiotic concentration and change in incubation temperature play a crucial role in the lactic coagulation of yogurt, such that the best lactic coagulation was observed at 50°C and a concentration of 50ppb. Hence, for tetracycline concentrations less than 75ppb, a process temperature of 50°C and incubation time of ~10 h recommend for fermentation of milk containing antibiotics.

Keywords: antibiotics residues, yogurt, fermentation parameters, incubation temperature

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Authors: Mirosław Krzemieniewski, Marcin Zieliński, Marcin Dębowski

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

Keywords: depolymerisation, pre-treatment, biomass, fermentation

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Authors: Wizna, Helmi Muis, Hafil Abbas

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

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

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

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

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

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

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

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

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Authors: Anuoluwapo O. Adewole

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

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

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

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

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

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Authors: Jean Paul Hategekimana, Josiane Irakoze, Eugene Niyonzima, Annick Ndekezi

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Cassava (Manihot esculenta Crantz) is a root crop comprising an anti-nutritive factor known as cyanide. The compound can be removed by numerous processing methods such as boiling, fermentation, blanching, and sun drying to avoid the possibility of cyanide poisoning. Inappropriate processing mean can lead to disease and death. Cassava-based dishes are consumed in different ways, where cassava is cultivated according to their culture and preference. However, they have been shown to be unsafe based on high cyanide levels. The current study targeted to resolve the problem of high cyanide in cassava consumed in Rwanda. This study was conducted to determine the effect of slicing, blanching, and soaking time to reduce the fermentation duration of cassava for hydrogen cyanide (HCN) in mg/g removal. Cassava was sliced into three different portions (1cm, 2cm, and 5cm). The first portions were naturally fermented for seven days, where each portion was removed every 24 hours from soaking tanks and then oven dried at a temperature of 60°C and then milled to obtain naturally fermented cassava flours. Other portions of 1cm, 2cm, and 5cm were blanched for 2, 5, 10 min, respectively, and each similarly dried at 60°C and milled to produce blanched cassava flour. Other blanched portions were used to follow the previous fermentation steps. The last portions, which formed the control, were simply chopped. Cyanide content and starch content in mg/100g were investigated. According to the conducted analysis on different cassava treatments for detoxification, found that usual fermentation can be used, but for sliced portions aimed to size reduction for the easy hydrogen cyanide diffuse out and it takes four days to complete fermentation, which has reduced at 94.44% with significantly different (p<0.05)of total hydrogen cyanide contained in cassava to safe level of consumption, and what is recommended as more effective is to apply blanching combined with fermentation due to the fact that, it takes three days to complete hydrogen cyanide removal at 95.56% on significantly different (p<0.05) of reduction to the safe level of consumption.

Keywords: cassava, cyanide, blanching, drying, fermentation

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

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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: dates, ethanol, fructose, fermentation, S. cerevisiae

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Authors: Ife O. Bolaji

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Environmental concerns arising from the use of fossil fuels has increased the interest in the development of renewable and sustainable sources of energy. This would minimize the dependence on fossil fuels and serve as future alternatives. Organic wastes contain carbohydrates, proteins and lipids, which can be utilised as carbon sources for the production of bio-based products. Cellulose is the most abundant natural biopolymer, being the main structural component of lignocellulosic materials. The aim of this project is to develop a biological process for the hydrolysis and fermentation of organic wastes into ethanol and organic acids. The hydrolysis and fermentation processes are integrated in a single vessel using undefined mixed culture microorganisms. The anaerobic fermentation of microcrystalline cellulose was investigated in continuous and batch reactors at 25°C with an appropriate growth medium for cellulase formation, hydrolysis, and fermentation. The reactors were inoculated with soil (B1, C1, C3) or sludge from an anaerobic digester (B2, C2) and the breakdown of cellulose was monitored by measuring the production of ethanol, organic acids and the residual cellulose. The batch reactors B1 and B2 showed negligible microbial activity due to inhibition while the continuous reactors, C1, C2 and C3, exhibited little cellulose hydrolysis which was concealed by the cellulose accumulation in the reactor. At the end of the continuous operation, the reactors C1, C2 and C3 were operated under batch conditions. 48%, 34% and 42% cellulose had been fermented by day 88, 55 and 55 respectively of the batch fermentation. Acetic acid, ethanol, propionic acid and butyric acids were the main fermentation products in the reactors. A stable concentration of 0.6 g/l ethanol and 5 g/L acetic acid was maintained in C3 for several weeks due to reduced activity of methanogens caused by the decrease in pH. Thus far, the results have demonstrated that mixed microbial culture is capable of hydrolysing and fermenting cellulose under lenient conditions. The fermentation of cellulose has been found effective in a combination of continuous and batch processes.

Keywords: cellulose, hydrolysis, mixed culture, organic waste

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

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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: defaunation, nitrate, fermentation, methane production

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