Search results for: fermentation parameters

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: Calister Wingang Makebe, Wilson Agwanande Ambindei, Zangue Steve Carly Desobgo, Abraham Billu, Emmanuel Jong Nso, P. Nisha

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Traditional liquid-state fermentation processes of Annona muricata L. juice can result in fluctuating product quality and quantity due to difficulties in control and scale up. This work describes a laboratory-scale batch fermentation process to produce a probiotic Annona muricata L. enzymatically extracted juice, which was modeled using the Doehlert design with independent extraction factors being incubation time, temperature, and enzyme concentration. It aimed at a better understanding of the traditional process as an initial step for future optimization. Annona muricata L. juice was fermented with L. acidophilus (NCDC 291) (LA), L. casei (NCDC 17) (LC), and a blend of LA and LC (LCA) for 72 h at 37 °C. Experimental data were fitted into mathematical models (Monod, Logistic and Luedeking and Piret models) using MATLAB software, to describe biomass growth, sugar utilization, and organic acid production. The optimal fermentation time was obtained based on cell viability, which was 24 h for LC and 36 h for LA and LCA. The model was particularly effective in estimating biomass growth, reducing sugar consumption, and lactic acid production. The values of the determination coefficient, R2, were 0.9946, 0.9913 and 0.9946, while the residual sum of square error, SSE, was 0.2876, 0.1738 and 0.1589 for LC, LA and LCA, respectively. The growth kinetic parameters included the maximum specific growth rate, µm, which was 0.2876 h-1, 0.1738 h-1 and 0.1589 h-1, as well as the substrate saturation, Ks, with 9.0680 g/L, 9.9337 g/L and 9.0709 g/L respectively for LC, LA and LCA. For the stoichiometric parameters, the yield of biomass based on utilized substrate (YXS) was 50.7932, 3.3940 and 61.0202, and the yield of product based on utilized substrate (YPS) was 2.4524, 0.2307 and 0.7415 for LC, LA, and LCA, respectively. In addition, the maintenance energy parameter (ms) was 0.0128, 0.0001 and 0.0004 with respect to LC, LA and LCA. With the kinetic model proposed by Luedeking and Piret for lactic acid production rate, the growth associated and non-growth associated coefficients were determined as 1.0028 and 0.0109, respectively. The model was demonstrated for batch growth of LA, LC, and LCA in Annona muricata L. juice. The present investigation validates the potential of Annona muricata L. based medium for heightened economical production of a probiotic medium.

Keywords: L. acidophilus, L. casei, fermentation, modelling, kinetics

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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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Authors: G. A. Amin, A. D. Al-Talhi

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The bacterium B. subtilis produced surfactin in conventional batch culture as a growth associated product and a growth rate (0.4 h-1). A fed-batch process was developed and the fermentative substrate and other nutrients were fed on hourly basis and according to the growth rate of the bacterium. Conversion of different quantities of Maldex-15 into surfactin was investigated in five different fermentation runs. In all runs, most of Maldex-15 was consumed and converted into surfactin and cell biomass with appreciable efficiencies. The best results were obtained with fermentation run supplied with 200 g Maldex-15. Up to 35.4 g.l-1 of surfactin and cell biomass of 30.2 g.l-1 were achieved in 12 hrs. Also, markedly substrate yield of 0.269 g/g and volumetric reactor productivity of 2.61 g.1-1.h-1 were obtained confirming the establishment of a cost effective commercial surfactin production.

Keywords: Bacillus subtilis, biosurfactant, exponentially fed-batch fermentation, surfactin

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Authors: M. Jerold, V. Sivasubramanian, A. George, B.S. Ashik, S. S. Kumar

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Bioethanol is a sustainable biofuel that can be used alternative to fossil fuels. Today, third generation (3G) biofuel is gaining more attention than first and second-generation biofuel. The more lignin content in the lignocellulosic biomass is the major drawback of second generation biofuels. Algae are the renewable feedstock used in the third generation biofuel production. Algae contain a large number of carbohydrates, therefore it can be used for the fermentation by hydrolysis process. There are two groups of Algae, such as micro and macroalgae. In the present investigation, Macroalgae was chosen as raw material for the production of bioethanol. Two marine algae viz. Ulva Lactuca and Sargassum swartzii were used for the experimental studies. The algal biomass was characterized using various analytical techniques like Elemental Analysis, Scanning Electron Microscopy Analysis and Fourier Transform Infrared Spectroscopy to understand the physio-Chemical characteristics. The batch experiment was done to study the hydrolysis and operation parameters such as pH, agitation, fermentation time, inoculum size. The saccharification was done with acid and alkali treatment. The experimental results showed that NaOH treatment was shown to enhance the bioethanol. From the hydrolysis study, it was found that 0.5 M Alkali treatment would serve as optimum concentration for the saccharification of polysaccharide sugar to monomeric sugar. The maximum yield of bioethanol was attained at a fermentation time of 9 days. The inoculum volume of 1mL was found to be lowest for the ethanol fermentation. The agitation studies show that the fermentation was higher during the process. The percentage yield of bioethanol was found to be 22.752% and 14.23 %. The elemental analysis showed that S. swartzii contains a higher carbon source. The results confirmed hydrolysis was not completed to recover the sugar from biomass. The specific gravity of ethanol was found to 0.8047 and 0.808 for Ulva Lactuca and Sargassum swartzii, respectively. The purity of bioethanol also studied and found to be 92.55 %. Therefore, marine algae can be used as a most promising renewable feedstock for the production of bioethanol.

Keywords: algae, biomass, bioethaol, biofuel, pretreatment

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Authors: Young-Ran Song, Byeong-Uk Lim, Sang-Ho Baik

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This study was initiated in order to develop a method for soy sauce fermentation at low salt concentrations without decreasing quality. Soy sauce was fermented with the fermentation starter (meju) and different salt contents (8-14%, w/v) by inoculating two strains or not, in which Torulaspora delbrueckii and Pichia guilliermondii strains having different abilities to induce sterilizing effects or enhance flavor production were used. As the results, there were microbial and biochemical differences among prepared soy sauce. First, Staphylococcus and Enterococcus spp. in addition to Bacillus genus that is the most important bacteria in Korean fermented soy product were detected by salt reduction. However, application of yeast starters can inhibit the undesirable bacterial growth. Moreover, PCA bi-plots of major principal components on various biochemical parameters (final pH, total acidity, soluble sugar, reducing sugar, ethanol and 32 volatile flavor compounds) were drawn to demonstrate the physicochemical differences and similarities among the samples. It was confirmed that the soy sauce samples produced with different salt concentrations were clearly different since salt reduction induced low contents of acids, alcohols and esters with higher acidity. However despite low salt concentration, combining two different yeasts appeared to have similar characteristics to the high salt-fermented soy sauce with elevated concentrations of ethanol, some alcohols, and most ketones, hence resulted in a balance of more complex and richer flavors with a flavor profile pattern identical to that of high-salt.

Keywords: Soy sauce, low salt, fermentation, yeast.

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Authors: Valentina Obradović, Josip Mesić, Maja Ergović Ravančić, Kamila Mijowska, Brankica Svitlica

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French oak and American oak barrels are most famous all over the world, but barrels of different origin can also be used for obtaining high quality wines. The aim of this research was to compare the influence of different Slavonian (Croatian) and French oak barrels on the quality of Chardonnay wine. Grapes were grown in Croatian wine growing region of Kutjevo in 2015. Chardonnay wines were tested for basic oenological parameters (alcohol, extract, reducing sugar, SO2, acidity), total polyphenols content (Folin-Ciocalteu method), antioxidant activity (ABTS and DPPH method) and color density. Sensory evaluation was performed by students of viticulture/oenology. Samples produced by classical fermentation and ageing in French oak barrels, had better results for polyphenols and sensory evaluation (especially low toasting level) than samples in Slavonian barrels. All tested samples were scored as a “quality” or “premium quality” wines. Sur lie method of fermentation and ageing in Slavonian oak barrel had very good extraction of polyphenols and high antioxidant activity with the usage of authentic yeasts, while commercial yeast strain resulted in worse chemical and sensory parameters.

Keywords: chardonnay, French oak, Slavonian oak, sur lie

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Authors: Diya Alsafadi, Fawwaz Khalili, Mohammad W. Amer

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Butanol is an important industrial solvent and potentially a better liquid transportation biofuel than ethanol. The cost of feedstock is one key problem associated with the biobutanol production. Date palm is sugar-rich fruit and highly abundant. Thousands of tons of date wastes that generated from date processing industries are thrown away each year and imposing serious environmental problems. To exploit the utilization of renewable biomass feedstock, date palm waste was utilized for butanol production by Clostridium acetobutylicum DSM 1731. Fermentation conditions were optimized by investigating several parameters that affect the production of butanol such as temperature, substrate concentration and pH. The highest butanol yield (1.0 g/L) and acetone, butanol, and ethanol (ABE) content (1.3 g/L) were achieved at 20 g/L date waste, pH 5.0 and 37 °C. These results suggest that date palm waste can be used for biobutanol production.

Keywords: biofuel, acetone-butanol-ethanol fermentation, date palm waste, Clostridium acetobutylicum

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Authors: Henry O. Udeha, Kwaku G. Duodub, Afam I. O. Jideanic

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Finger millet (FM) [Eleusine coracana] is considered as a potential ‘‘super grain’’ by the United States National Academies as one of the most nutritious among all the major cereals. The regular consumption of FM-based diets has been associated with reduced risk of diabetes, cataract and gastrointestinal tract disorder. Hyperglycaemic, hypocholesterolaemic and anticataractogenic, and other health improvement properties have been reported. This study examined the effect of fermentation time and microbial source on physicochemical properties, phenolic compounds and antioxidant activity of two finger millet (FM) malt flours. Sorghum was used as an external reference. The grains were malted, mashed and fermented using the grain microflora and Lactobacillus fermentum. The phenolic compounds of the resulting beverage were identified and quantified using ultra-performance liquid chromatography (UPLC) and mass spectrometer system (MS). A fermentation-time dependent decrease in pH and viscosities of the beverages, with a corresponding increase in sugar content were noted. The phenolic compounds found in the FM beverages were protocatechuic acid, catechin and epicatechin. Decrease in total phenolics of the beverages was observed with increased fermentation time. The beverages exhibited 2, 2-diphenyl-1-picrylhydrazyl, 2, 2՛-azinobis-3-ethylbenzthiazoline-6-sulfonic acid radical scavenging action and iron reducing activities, which were significantly (p < 0.05) reduced at 96 h fermentation for both microbial sources. The 24 h fermented beverages retained a higher amount of total phenolics and had higher antioxidant activity compared to other fermentation periods. The study demonstrates that FM could be utilised as a functional grain in the production of non-alcoholic beverage with important phenolic compounds for health promotion and wellness.

Keywords: antioxidant activity, eleusine coracana, fermentation, phenolic compounds

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Authors: Tuaumelsan Shumye Gebre, Shimelis Admassu Emire, Simon Okomo Aloo, Ramachandran Chelliah, Deog-Hwan Oh

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This study investigates the fermentation of cereal substrates, based on the Ethiopian traditional beverage borde, using probiotic strains of Pediococcus acidilactici WS07 and Saccharomyces cerevisiae AM18 used singly and in co-culture. The pH and titratable acidity, microbial growth dynamics, fermentable sugars profile, volatile organic compounds, total flavonoid content, total phenolic content, antioxidant activity, pancreatic lipase, and α-glucosidase inhibition were analyzed. The viability of every tested strain remained higher than 7 log CFU/mL, satisfying the requirements suggested for probiotic food items. The formation of organic acids is what caused the pH to decrease from roughly 6.6 to 3.8, yet this had no effect on the viability of the microorganisms. The fermentation process, involving P. acidilactici WS07 and S. cerevisiae AM18, led to the utilization of initial carbohydrates, production of organic acids, and generation of volatile compounds that enhance flavor and aroma. Ethanol and glycerol concentrations increased during fermentation, particularly in co-culture assays, contributing to the sensory qualities and stability of the beverages. The primary organic acids generated during fermentation were lactic and acetic acids. A total of 22 volatile substances, such as acids, alcohols, aldehydes, esters, ketones, and other substances, were found. Furthermore, the study demonstrates that fermentation of maize and sorghum with P. acidilactici WS07 and S. cerevisiae AM18 enhances the antioxidant activity and inhibition of pancreatic lipase and α-glucosidase, suggesting potential benefits in managing obesity and diabetes. Therefore, co-cultivating S. cerevisiae AM18 and P. acidilactici WS07 in cereal fermentation led to the successful production of probiotic drinks.

Keywords: probiotic beverage, Pediococcus acidilactici, Saccharomyces cerevisiae, volatile compounds

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Authors: Thi Huong Vu, Haelee Fenton, Thi Huong Tra Nguyen, Gary Dykes

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In the present study, a submerged fermentation of carob kibble with Saccharomyces cerevisiae (S. cerevisiae) was performed. The total phenolic content and antioxidant activity in fermented carob kibble were determined by Folin–Ciocalteu method and scavenging capacity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS). The study showed that S. cerevisiae improved total phenolic content by 45 % and 50 % in acetone and water extracts respectively. Similarly, the antioxidant capacity of water extracts increased by 25 % and 41%, while acetone extracts indicated by 70% and 80% in DPPH and ABTS respectively. It is also found that initial pH 7.0 was more effective in improvement of total phenolic content and antioxidant activity. The efficiency of treatment was recorded at 15 h. This report suggested that submerged fermentation with S. cerevisiae is a potential and cost effective manner to further increase bioactive compounds in carob kibble, which are in use for food, cosmetic and pharmaceutical industries.

Keywords: antioxidant activity, carob kibble, saccharomyces cerevisiae, submerged fermentation, total phenolics

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

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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, malting, ratio, roasting, wettability

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

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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, African yam bean, acha, biscuits, meat-pie

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Authors: Neti Yuliana, Srisetyani, Siti Nurdjanah, Dewi Sartika, Yoan Martiansari, Putri Nabila

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Naturally sweet potato flour usually requires a modification process to improve its inherent property for expanding its application in food system. The study was aimed to modify sweet potato flour (SPF), to increase its utilization for composite noodle production, trough fermentation of sweet potato slices before its flouring process. Fermentation were prepared with five different starters: pickle brine, Lactobacillus plantarum, Leuconostoc mesenteroides, mixed of Lactobacillus plantarum, Leuconostoc mesenteroides , and mixed of Lactobacillus plantarum, Leuconostoc mesenteroides, and Sacharomyces cerevisiae. Samples were withdrawn every 0, 24, 48, 72 and 96 hours. The fermented flours were characterized for swelling power, solubility, paste transmittance, pH, sensory properties (acidic aroma and whiteness), and the amount of broken composite noodle strips. The results indicated that there was no significant effect of different starters on fermented SPF characteristic and on the amount of broken noodle strip, while length of fermentation significantly affected. Longer fermentation, reaching 48-72 h, increased swelling power, pH, acidic aroma and whiteness of flour and reduced solubility, paste transmittance, and the amount of broken noodle strip. The results suggested that fermentation within 48-72 h period of time could provide great composite SPF for noodle.

Keywords: starters, fermented flour, sweet potato, composite noodle

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

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

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

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Authors: Zelal Polat, Şebnem Harsa, Semra Ülkü

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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: fermentation, ion exchange, lactic acid, purification, whey

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Authors: Noura El-Ahmady El-Naggar

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L-asparaginase is an important enzyme as therapeutic agents used in combination therapy with other drugs in the treatment of acute lymphoblastic leukemia in children. L-asparaginase producing actinomycete strain, NEAE-K, was isolated from soil sample and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as new subsp. Streptomyces rochei subsp. chromatogenes NEAE-K and sequencing product (1532 bp) was deposited in the GenBank database under accession number KJ200343. The study was conducted to screen parameters affecting the production of L-asparaginase by Streptomyces rochei subsp. chromatogenes NEAE-K on solid state fermentation using Plackett–Burman experimental design. Sixteen different independent variables including incubation time, moisture content, inoculum size, temperature, pH, soybean meal+ wheat bran, dextrose, fructose, L-asparagine, yeast extract, KNO3, K2HPO4, MgSO4.7H2O, NaCl, FeSO4. 7H2O, CaCl2, and three dummy variables were screened in Plackett–Burman experimental design of 20 trials. The most significant independent variables affecting enzyme production (dextrose, L-asparagine and K2HPO4) were further optimized by the central composite design. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase by Streptomyces rochei subsp. chromatogenes NEAE-K from solid state fermentation: g/L (soybean meal+ wheat bran 15, dextrose 3, fructose 4, L-asparagine 8, yeast extract 2, KNO3 1, K2HPO4 2, MgSO4.7H2O 0.5, NaCl 0.1, FeSO4. 7H2O 0.02, CaCl2 0.01), incubation time 7 days, moisture content 50%, inoculum size 3 mL, temperature 30°C, pH 8.5.

Keywords: streptomyces rochei subsp. chromatogenes neae-k, 16s rrna, identification, solid state fermentation, l-asparaginase production, plackett-burman design, central composite design

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Authors: Calister Wingang Makebe, Wilson Ambindei Agwanande, Emmanuel Jong Nso, P. Nisha

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Traditional liquid-state fermentation processes of Annona muricata L. juice can result in fluctuating product quality and quantity due to difficulties in control and scale up. This work describes a laboratory-scale batch fermentation process to produce a probiotic Annona muricata L. enzymatically extracted juice, which was modeled using the Doehlert design with independent extraction factors being incubation time, temperature, and enzyme concentration. It aimed at a better understanding of the traditional process as an initial step for future optimization. Annona muricata L. juice was fermented with L. acidophilus (NCDC 291) (LA), L. casei (NCDC 17) (LC), and a blend of LA and LC (LCA) for 72 h at 37 °C. Experimental data were fitted into mathematical models (Monod, Logistic and Luedeking and Piret models) using MATLAB software, to describe biomass growth, sugar utilization, and organic acid production. The optimal fermentation time was obtained based on cell viability, which was 24 h for LC and 36 h for LA and LCA. The model was particularly effective in estimating biomass growth, reducing sugar consumption, and lactic acid production. The values of the determination coefficient, R2, were 0.9946, 0.9913 and 0.9946, while the residual sum of square error, SSE, was 0.2876, 0.1738 and 0.1589 for LC, LA and LCA, respectively. The growth kinetic parameters included the maximum specific growth rate, µm, which was 0.2876 h-1, 0.1738 h-1 and 0.1589 h-1 as well as the substrate saturation, Ks, with 9.0680 g/L, 9.9337 g/L and 9.0709 g/L respectively for LC, LA and LCA. For the stoichiometric parameters, the yield of biomass based on utilized substrate (YXS) was 50.7932, 3.3940 and 61.0202, and the yield of product based on utilized substrate (YPS) was 2.4524, 0.2307 and 0.7415 for LC, LA, and LCA, respectively. In addition, the maintenance energy parameter (ms) was 0.0128, 0.0001 and 0.0004 with respect to LC, LA and LCA. With the kinetic model proposed by Luedeking and Piret for lactic acid production rate, the growth associated, and non-growth associated coefficients were determined as 1.0028 and 0.0109, respectively. The model was demonstrated for batch growth of LA, LC, and LCA in Annona muricata L. juice. The present investigation validates the potential of Annona muricata L. based medium for heightened economical production of a probiotic medium.

Keywords: L. acidophilus, L. casei, fermentation, modelling, kinetics

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Authors: Antonietta Maoloni, Federica Cardinali, Vesna Milanović, Andrea Osimani, Ilario Ferrocino, Maria Rita Corvaglia, Luca Cocolin, Lucia Aquilanti

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Table olives (Olea europaea L.) are among the most important fermented vegetables all over the world, while sea fennel (Crithmum maritimum L.) is an emerging food crop with interesting nutritional and sensory traits. Both of them are characterized by the presence of several bioactive compounds with potential beneficial health effects, thus representing two valuable substrates for the manufacture of innovative vegetable-based preserves. Given these premises, the present study was aimed at exploring the co-fermentation of table olives and sea fennel to produce new high-value preserves. Spanish style or Greek style processing method and the use of a multiple strain starter were explored. The preserves were evaluated for their microbial dynamics and key sensory traits. During the fermentation, a progressive pH reduction was observed. Mesophilic lactobacilli, mesophilic lactococci, and yeasts were the main microbial groups at the end of the fermentation, whereas Enterobacteriaceae decreased during fermentation. An evolution of the microbiota was revealed by metataxonomic analysis, with Lactiplantibacillus plantarum dominating in the late stage of fermentation, irrespective of processing method and use of the starter. Greek style preserves resulted in more crunchy and less fibrous than Spanish style one and were preferred by trained panelists.

Keywords: lactic acid bacteria, Lactiplantibacillus plantarum, metataxonomy, panel test, rock samphire

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Authors: Shraddha Digole, Swarali Hingse, Uday Annapure

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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: Box-Behnken design, fermentation, mycophenolic acid, Penicillium brevicompactum

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Authors: Alexandra Tucaliuc, Alexandra C. Blaga, Anca I. Galaction, Lenuta Kloetzer, Dan Cascaval

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Ergosterol (ergosta-5,7,22-trien-3β-ol), also known as provitamin D2, is the precursor of vitamin D2 (ergocalciferol), because it is converted under UV radiation to this vitamin. The natural sources of ergosterol are mainly the yeasts (Saccharomyces sp., Candida sp.), but it can be also found in fungus (Claviceps sp.) or plants (orchids). In the yeasts cells, ergosterol is accumulated in membranes, especially in free form in the plasma membrane, but also as esters with fatty acids in membrane lipids. The chemical synthesis of ergosterol does not represent an efficient method for its production, in these circumstances, the most attractive alternative for producing ergosterol at larger-scale remains the aerobic fermentation using S. cerevisiae on glucose or by-products from agriculture of food industry as substrates, in batch or fed-batch operating systems. The aim of this work is to analyze comparatively the influence of aeration efficiency on ergosterol production by S. cerevisiae in batch and fed-batch fermentations, by considering different levels of mixing intensity, aeration rate, and n-dodecane concentration. The effects of the studied factors are quantitatively described by means of the mathematical correlations proposed for each of the two fermentation systems, valid both for the absence and presence of oxygen-vector inside the broth. The experiments were carried out in a laboratory stirred bioreactor, provided with computer-controlled and recorded parameters. n-Dodecane was used as oxygen-vector and the ergosterol content inside the yeasts cells has been considered at the fermentation moment related to the maximum concentration of ergosterol, 9 hrs for batch process and 20 hrs for fed-batch one. Ergosterol biosynthesis is strongly dependent on the dissolved oxygen concentration. The hydrocarbon concentration exhibits a significant influence on ergosterol production mainly by accelerating the oxygen transfer rate. Regardless of n-dodecane addition, by maintaining the glucose concentration at a constant level in the fed-batch process, the amount of ergosterol accumulated into the yeasts cells has been almost tripled. In the presence of hydrocarbon, the ergosterol concentration increased by over 50%. The value of oxygen-vector concentration corresponding to the maximum level of ergosterol depends mainly on biomass concentration, due to its negative influences on broth viscosity and interfacial phenomena of air bubbles blockage through the adsorption of hydrocarbon droplets–yeast cells associations. Therefore, for the batch process, the maximum ergosterol amount was reached for 5% vol. n-dodecane, while for the fed-batch process for 10% vol. hydrocarbon.

Keywords: bioreactors, ergosterol, fermentation, oxygen-vector

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Authors: Scholz Heiko, Kuhne Petra, Heckenberger Gerd

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

Keywords: suckler cow, feeding systems, crude fiber, digestibilty of organic matter

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

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Given the high-utility of peroxidase, its production in large amount is of utmost importance. Over the years, actinomycetes have been the major peroxidase-producing bacteria. Consequently, other classes of bacteria with peroxidase production potentials are underexplored. This study, therefore, sought to enhance peroxidase production by a Raoultella species, a new ligninolytic proteobacteria strain, by determining the optimum culture conditions (initial pH, incubation temperature and agitation speed) for peroxidase production under submerged fermentation using the classical process of one variable at a time and supplementing the fermentation medium with some lignin model and inorganic nitrogen compounds. Subsequently, the time-course assay was carried out under optimized conditions. Then, some agricultural residues were valorized for peroxidase production under solid state fermentation. Peroxidase production was optimal at initial pH 5, incubation temperature of 35 °C and agitation speed of 150 rpm with guaiacol and ammonium chloride as the best inducer and nitrogen supplement respectively. Peroxidase production by the Raoultella species was optimal at 72 h with specific productivity of 16.48 ± 0.89 U mg⁻¹. A simultaneous production of a non-peroxide dependent extracellular enzyme which suggests probable laccase production was observed with specific productivity of 13.63 ± 0.45 U mg⁻¹ while sawdust gave the best peroxidase yield under solid state fermentation. In conclusion, peroxidase production by the Raoultella species was increased by 3.40-fold.

Keywords: enzyme production, ligninolytic bacteria, peroxidase, proteobacteria

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

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The effects of electro-stimulation of S. cerevisiae cells in colloidal suspension by Pulsed Electric Fields ‎‎(PEF) with electric field strength E = 20 – 2000 V.cm-1 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 V.cm-1) 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: intensification, yeast, fermentation, electroporation, biotechnology

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Authors: Marcin Dębowski, Marcin Zieliński, Mirosław Krzemieniewski

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This publication presents a reactor designed for methane fermentation of organic substrates. The design is based on rotating cellular cylinders connected to a biomass feeder and an ultrasonic generator. This allows for simultaneous mixing and partial disintegration of the biomass, as well as stimulating higher metabolic rates within the microorganisms. Such a design allows from 2-fold to 14-fold reduction of power usage when compared to conventional mixing systems. The sludge does not undergo mechanical deformation during the mixing process, which improves substrate biodegradation efficiency by 10-15%. Cavitation occurs near the surface of the rods, partially releasing the biomass and separating it from the destroyed microorganisms. Biogas is released further away from the cellular cylinder rods due to the effect of the ultrasonic waves, in addition to increased biochemical activity of the microorganisms and increased exchange of the nutrient medium with metabolic products, which results in biogas production increase by about 15%.

Keywords: methane fermentation, bioreactors, biomass, mixing system

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Authors: Regiane A. Oliveira, Carlos E. Vaz Rossell, Rubens Maciel Filho

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Lactic acid, besides a valuable chemical may be considered a platform for other chemicals. In fact, the feasibility of hemicellulosic sugars as feedstock for lactic acid production process, may represent the drop of some of the barriers for the second generation bioproducts, especially bearing in mind the 5-carbon sugars from the pre-treatment of sugarcane bagasse. Bearing this in mind, the purpose of this study was to use the hemicellulosic liquor from sugarcane bagasse as a substrate to produce lactic acid by fermentation. To release of sugars from hemicellulose it was made a pre-treatment with a diluted sulfuric acid in order to obtain a xylose's rich liquor with low concentration of inhibiting compounds for fermentation (≈ 67% of xylose, ≈ 21% of glucose, ≈ 10% of cellobiose and arabinose, and around 1% of inhibiting compounds as furfural, hydroxymethilfurfural and acetic acid). The hemicellulosic sugars associated with 20 g/L of yeast extract were used in a fermentation process with Lactobacillus plantarum to produce lactic acid. The fermentation process pH was controlled with automatic injection of Ca(OH)2 to keep pH at 6.00. The lactic acid concentration remained stable from the time when the glucose was depleted (48 hours of fermentation), with no further production. While lactic acid is produced occurs the concomitant consumption of xylose and glucose. The yield of fermentation was 0.933 g lactic acid /g sugars. Besides, it was not detected the presence of by-products, what allows considering that the microorganism uses a homolactic fermentation to produce its own energy using pentose-phosphate pathway. Through facultative heterofermentative metabolism the bacteria consume pentose, as is the case of L. plantarum, but the energy efficiency for the cell is lower than during the hexose consumption. This implies both in a slower cell growth, as in a reduction in lactic acid productivity compared with the use of hexose. Also, L. plantarum had shown to have a capacity for lactic acid production from hemicellulosic hydrolysate without detoxification, which is very attractive in terms of robustness for an industrial process. Xylose from hydrolyzed bagasse and without detoxification is consumed, although the hydrolyzed bagasse inhibitors (especially aromatic inhibitors) affect productivity and yield of lactic acid. The use of sugars and the lack of need for detoxification of the C5 liquor from sugarcane bagasse hydrolyzed is a crucial factor for the economic viability of second generation processes. Taking this information into account, the production of second generation lactic acid using sugars from hemicellulose appears to be a good alternative to the complete utilization of sugarcane plant, directing molasses and cellulosic carbohydrates to produce 2G-ethanol, and hemicellulosic carbohydrates to produce 2G-lactic acid.

Keywords: fermentation, lactic acid, hemicellulosic sugars, sugarcane

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

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

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

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Authors: Marianna Ciaccia, Gennaro Agrimi, Isabella Pisano, Maurizio Bettiga, Silvia Rapacioli, Giulia Mensa, Monica Marzagalli

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Objective: Untargeted metabolomic analysis of extracellular metabolites is a powerful approach that focuses on comprehensively profiling in the extracellular space. In this study, we applied extracellular metabolomic analysis to investigate the metabolism of two probiotic microorganisms with health benefits that extend far beyond the digestive tract and the immune system. Methods: Analytical techniques employed in extracellular metabolomic analysis encompass various technologies, including mass spectrometry (MS), which enables the identification of metabolites present in the fermentation media, as well as the comparison of metabolic profiles under different experimental conditions. Multivariate statistical analysis techniques like principal component analysis (PCA) or partial least squares-discriminant analysis (PLS-DA) play a crucial role in uncovering metabolic signatures and understanding the dynamics of metabolic networks. Results: Different types of supernatants from fermentation processes, such as dairy-free, not dairy-free media and media with no cells or pasteurized, were subjected to metabolite profiling, which contained a complex mixture of metabolites, including substrates, intermediates, and end-products. This profiling provided insights into the metabolic activity of the microorganisms. The integration of advanced software tools has facilitated the identification and characterization of metabolites in different fermentation conditions and microorganism strains. Conclusions: In conclusion, untargeted extracellular metabolomic analysis, combined with software tools, allowed the study of the metabolites consumed and produced during the fermentation processes of probiotic microorganisms. Ongoing advancements in data analysis methods will further enhance the application of extracellular metabolomic analysis in fermentation research, leading to improved bioproduction and the advancement of sustainable manufacturing processes.

Keywords: biotechnology, metabolomics, lactic bacteria, probiotics, postbiotics

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