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Republic of Srpska (RS) is one of the two entities in Bosnia and Herzegovina (BiH). It is located in South-eastern Europe, in the western part of the Balkan Peninsula. As in most developing countries, waste management is based on landfilling. The constant increase in waste generation creates a growing burden on the existing waste management system and the environment. Comparing the amount of waste with GDP, despite a much lower standard, RS/BiH residents, compared to EU residents, generate far more waste for the same amount of money. All generated waste is disposed of in landfills, which causes greenhouse gas emissions and has a negative effect on the environment. On the other side, untreated waste deposited in landfills represents the loss of potential of waste as a renewable energy source. In this study, an attempt has been made to represent the possibilities of sustainable waste-to-energy management in RS with the goal to provide reliable support to decision makers to direct waste management in a sustainable framework.

The aim of this study was to examine the effects of variation of organic (yeast extract, soybean flour and peptone) and inorganic nitrogen sources (NaNO2, KNO3 and (NH4)2HPO4) in medium for biobactericides production by Bacillus subtilis ATCC 6633. The antibacterial activity of samples against the reference strain Xanthomonas campestris ATCC 13951 and different Xanthomonas strains isolated from cabbage, kale and cauliflower was determined in vitro by discdiffusion method. Following statistical analysis, it was concluded that both nitrogen sources have statistically significant effect on biobactericides production under applied conditions whereby the influence of inorganic nitrogen is more pronounced. The results of Duncan's test suggested that the highest biosynthesis of antibacterial compounds was achieved in both media, with or without yeast extract, when NaNO2 was used as an inorganic nitrogen source. The results from this study can be used in further research to improve biobactericides production by the reference B. subtilis strain.

This study is concerned with the effect of different cultivation techniques on enzymes production from sugar beet pulp by strain Neurospora crassa isolated from the environment. Cultivation of selected producing microorganism was carried out under the same process conditions using five techniques. Bioprocess efficacy was estimated based on amylolytic, cellulolytic and xylanolytic activity of prepared enzymes mixtures. The obtained results indicate that the selection of cultivation technique had a statistically significant effect on the production of examined hydrolytic enzymes. It was confirmed that solid state cultivation with spontaneous aeration is the best cultivation technique for the production of amylolytic, cellulolytic and xylanolytic enzymes from sugar beet pulp by Neurospora crassa. Submerged cultivation of producing strain with spontaneous aeration resulted in the lowest production of all investigated enzymes under applied experimental conditions. The obtained results are the basis for further research aimed to increase the enzymes yield and activity of their mixture. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR-31002]

Xanthan is the most important microbial exopolysaccharide used in different branches of food and non-food industry. Due to high consumption of glucose and sucrose, usually utilized carbon sources in many bioprocesses, the most prominent research in xanthan production is related to the isolation of new xanthan producing strains capable to metabolize other carbon sources and to produce xanthan with characteristics appropriate for usage in food industry. The aim of this study was to examine the effect of variation of carbon sources in cultivation media on xanthan production by reference strain Xanthomonas campestris ATCC 13951 and five Xanthomonas strains isolated from infected pepper leaves. Bioprocess efficacy was estimated based on the quantity and quality of biosynthesized xanthan. The obtained results show that both, the selection of the carbon source and selection of producing strain have a statistically significant effect on bioprocess efficacy whereby the influence of carbon sources on the analyzed parameters is more pronounced. It was confirmed that Xanthomonas strains responsible for the production of a large amount of xanthan are not suitable for biosynthesis of good-quality biopolymer. Taking into account all results it can be concluded that the greatest potential in biotechnological xanthan production has strain PFNS PL4 and glucose or starch containing media which is the basis for further research.

Xanthan is a microbial biopolymer with a wide range of industrial applications and it is expected that the demand for this product will significantly increase in the coming decade and for this reason it is important to constantly work on improving all aspects of this biotechnological process. The aim of this research was to examine the kinetics of batch cultivation of Xanthomonas campestris ATCC 13951 using vegetable oil industry wastewaters as a basis for the cultivation medium, in order to produce the biopolymer xanthan. Kinetic modelling is very important for process control, reducing process costs and increasing product quality. By performing xanthan production on a medium with optimized content, the experimental values of content of biomass, carbon source and the desired product were obtained and used to determine the kinetics of biosynthesis. In order to describe biomass multiplication, product formation and carbon source consumption, the logistics, the Luedeking-Piret and modified Luedeking- -Piret equation, respectively, were successfully used. Additionally, using process simulation software (SuperPro Designer®), a process and cost model for a xanthan production facility was developed. The developed model represents the basis for a 21,294.29 and 23,107.97 kg/year xanthan production facility, which uses a vegetable oil industry wastewater-based medium and a semi-synthetic medium. The simulation model of the suggested xanthan production process, developed and based on defined kinetic models, represents an excellent basis for its further improvement and for increasing its efficiency.

Biotechnologically produced antifungal compounds for control of plant diseases caused by phytopathogenic fungi represent a promising alternative to synthetic pesticides. Fungi from the genera Alternaria and Fusarium are listed among important storage pathogens of apple fruits. A. alternata causes significant annual losses of apple fruit. Also, F. avenaceum is one of the most commonly encountered Fusarium species identified as the causal agent of a wet apple core rot. Species of the Streptomyces genera are soil bacteria that produce significant quantities of bioactive compounds in appropriate media. Defining the culture medium composition is the basis of bioprocesses development. Nitrogen source is critical component of cultivation medium and also the most useful tool for stimulation of the antifungal metabolites production. The aim of this study was to select the best nitrogen sources in medium for the production of antifungal compounds effective against A. alternata and F. avenaceum by S. hygroscopicus. Activity of the cultivation liquids on A. alternata and F. avenaceum isolates was tested in vitro using well diffusion method. The results in­dicate that maximum inhibition zone was reached in medium with soybean meal as nitrogen source with inhibition diameter more than 35 mm for both tested fungi. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR-31002]

In the effort to overcome the increase in antimicrobial resistance of different pathogens, natural products from microbial sources appear to be the most favorable alternative to current antibiotics. Production of antimicrobial compounds is highly dependent on the nutritional conditions. Hence, in order to achieve high product yields, selection of the media constituents and optimization of their concentrations are required. In this research, the possibility of antimicrobial substances production using Bacillus subtilis ATCC 6633 was investigated. Also, optimization of the cultivation medium composition in terms of contents of glycerol, sodium nitrite and phosphates was done. Response surface methodology and the method of desirability function were applied for determination of optimal values of the examined factors. The developed model predicts that the maximum inhibition zone diameters for Bacillus cereus ATCC 10876 (33.50 mm) and Pseudomonas aeruginosa ATCC 27853 (12.00 mm) are achieved when the initial contents of glycerol, sodium nitrite and phosphates were 43.72 g/L, 1.93 g/L and 5.64 g/L, respectively. The results of these experiments suggest that further research should include the utilization of crude glycerol as a carbon source and optimization of composition of such media and cultivation conditions in order to improve production of antimicrobial substances using Bacillus subtilis ATCC 6633.

The aim of this research was to examine the possibility of using the vegetable oil industry wastewater as the basis of the cultivation media for xanthan production, using Xanthomonas campestris ATCC 13951, in order to reuse the wastewater obtained in one industry as a raw material in another. Furthermore, the aim was to optimize the composition of wastewater based media in terms of carbon, nitrogen, and phosphorus content. Experiments were carried out in accordance with the Box-Behnken design with three factors on three levels (glucose: 10.00- -30.00 g/L; nitrogen: 0.02-0.20 g/L; phosphorus: 0.0045-0.045 g/L) and three repetitions at the central point. The success of biosynthesis was assessed by analysing broths to determine xanthan yield, cultivation broth apparent viscosity, residual carbon, nitrogen, and phosphorus content. Graphical and numerical optimization using response surface methodology was performed to determine the optimal content values of the carbon source, nitrogen, and phosphorus content. The results of graphical optimization suggest that for the defined response interval values, the defined model predicts that the xanthan production is possible when 15.00-16.00 g/L of the carbon source, 0.02-0.09 g/L of nitrogen and 0.01-0.02 g/L of phosphorus are added to the media that as a basis contains vegetable oil industry wastewater.

Wine industry generates large volumes of wastewaters resulting from numerous cleaning operations that occur during the different stages of winemaking. Disposal of these effluents in the environment causes huge problems due to their high organic and inorganic load and seasonal variability. The bioconversion of winery wastewaters in valuable product, such as xanthan, is an important alternative to overcome environmental problems. In this research, the possibility of xanthan production using Xanthomonas campestris on blended wastewaters from different stages of white and rose wine production with initial sugar content of 50 g/L was investigated. In addition to the media parameters (content of sugars, total and assimilable nitrogen, phosphorus, total dissolved salts and apparent viscosity), raw xanthan yield and degree of sugar conversion into product were determined in order to examine the success of xanthan biosynthesis. In applied experimental conditions, xanthan yield of 20.92 and 30.64 g/L and sugar conversion into product of 40.23 and 60.73% were achieved on wastewaters from white and rose wine production, respectively. The results of these experiments suggest that winery wastewaters, after additional optimization of the process in terms of the substrate composition and the cultivation conditions, may be a suitable raw material for industrial xanthan production.

The application of antifungal compounds produced by microorganisms in the control of plant diseases caused by phytopathogenic fungi is a promising alternative to synthetic pesticides. Among phytopathogenic fungi, Alternaria alternata and Fusarium avenaceum are significant pathogens responsible for the storage rot of apple fruits. During storage, transport and marketing A. alternata and F. avenaceum can cause significant losses of apple fruits and their control is of great importance for the producers and consumers. In the present study, the effects of agitation rate on the production of antifungal methabolite( s) by Streptomyces hygroscopicus in a 3-L lab-scale bioreactor (Biostat® Aplus, Sartorius AG, Germany) against two isolates of A. alternata and two isolates of F. avenaceum were investigated. The cultivation of S. hygroscopicus was carried out at 27°C with agitation rates of 100 rpm and 200 rpm during 7 days. The aim was to analyze the bioprocess parameters of biofungicide production in a medium containing glycerol as a carbon source, and examine the effect of agitation rate on the production of antifungal metabolite(s). The in vitro antifungal activity of the produced metabolites against fungi from the genera Alternaria and Fusarium grown on potato dextrose agar medium was determined every 24 h using wells technique. In the experiments conducted in the bioreactor at different stirring speeds, it was found that the maximum production of antifungal metabolites occurred after 96 hours of cultivation. A higher consumption of nutrients and a larger inhibition zone diameter was registered in the experiment with an agitation rate of 200 rpm. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR-31002]

Ethanol is an important industrial chemical with emerging potential as a biofuel to replace fossil fuels. In order to enhance the efficiency and yield of alcoholic fermentation, combined techniques such as cells immobilization and media optimization have been used. The aim of this study was the optimization of sodium alginate concentration and glucose and yeast extract content in the media for ethanol production with immobilized cells of Saccharomyces cerevisiae. Optimization of these parameters was attempted by using a Box-Behnken design using the response surface methodology. The obtained model predicts that the maximum ethanol content of 7.21% (v/v) is produced when the optimal values of sodium alginate concentration and initial content of glucose and yeast extract in the medium are 22.84 g/L, 196.42 g/L and 3.77 g/L, respectively. To minimize the number of yeast cells "eluted" from the alginate beads and residual glucose content in fermented media, additional two sets of optimization were made. The obtained results can be used for further techno-economic analyses of the process to select the optimum conditions of the fermentation process for industrial application.[Projekat Ministarstva nauke Republike Srbije, br. TR-31002]

The success of xanthan biosynthesis depends on several factors, most importantly the genetic potential of the production microorganism and cultivation media composition. Cultivation media composition affects the yield and quality of the desired product as well as production costs. This is why many studies focus on finding cheap alternative raw materials, especially carbon sources, to replace commercially used glucose and sucrose. In addition to the Xanthomonas campestris ATCC 13951 which is the primary industrial production microorganism, other Xanthomonas strains can produce xanthan as well. Under the same conditions, different strains produce different amounts of the biopolymer of varying quality. The aim of this paper is to compare producibility of phytopathogenic X. campestris strains, isolated from the environment with the reference X. campestris ATCC 13951 strain and to estimate the possibility of xanthan production using alternative glycerol-based media than the synthetic glucose-based media. Submerged cultivation on the medium based on glucose or glycerol (2.0 %w/v) was performed using the reference strain and eight isolated X. campestris strains. In order to assess the success of biosynthesis, xanthan yield and rheological properties were determined. Strains isolated from the environment produced yields between 2.98 g/L and 12.17 g/L on the glucose-based medium and 1.68 g/L and 6.31 g/L on the glycerol-based medium. Additionally, X. campestris ATCC 13951 provided the highest yield when using glucose (13.24 g/L), as well as glycerol-based medium (7.44 g/L). The obtained results indicate that in the applied experimental conditions and using all tested strains, glycerol is viable as a carbon source for the production of xanthan.

This study is concerned with the effect of different initial glycerol concentrations in the medium on xanthan production by Xanthomonas campestris ATCC 13951. Xanthan biosynthesis was carried out in batch mode under aerobic conditions at a temperature of 30oC and agitation rate of 150 rpm for 7 days. The process efficiency was estimated based on the values of raw xanthan yield, average molecular weight of the polymer and residual content of glycerol, total nitrogen and phosphorus. Based on these results, the initial concentration of glycerol as a carbon source in the production medium was suggested. In the applied experimental conditions, high raw xanthan yield (12.15 g/l) of good quality (Mw = 2.86•105 g/mol) and the lowest amount of residual nutrients (glycerol 2.75 g/l, nitrogen 0.46 g/l and phosphorus 0.67 g/l) was achieved in the medium with the initial glycerol content of 20 g/l. The obtained results are the basis for optimization of xanthan production on glycerol containing media in order to increase the product yield and quality.

The biocontrol agents are a very promising alternative to synthetic pesticides that are presently used to control plant diseases caused by phytopathogenic microorganisms. Members of the Bacillus genera are soil bacteria that produce significant quantities of agriculturally important bioactive compounds. Production of these compounds can be improved by changing the nutritional and environmental conditions. The aim of this study was the optimization of medium composition, using response surface methodology, for the production of compounds effective against Xanthomonas campestris ATCC 13951 by Bacillus subtilis ATCC 6633. To study the production of antimicrobial compounds by selected Bacillus strain, the producing microorganisms were cultivated on nutrient broth. The inhibition zone diameter of 18.0 mm obtained by the diffusion-disc method indicated that the used Bacillus subtilis strain produces compounds with antimicrobial activity against Xanthomonas campestris ATCC 13951. To optimize the composition of the cultivation medium in terms of glycerol, sodium nitrite and phosphates content, experiments were carried out in accordance with Box-Behnken design, and optimization of multiple responses was performed using the concept of desirability function. The developed model predicted that the maximum inhibition zone diameter (26.23 mm) against tested phytopathogen is achieved when the initial content of glycerol, sodium nitrite and phosphate were 50.00 g/L, 2.85 g/L and 11.00 g/L, respectively. To minimize the consumption of medium components and costs of effluents processing, additional optimization set was made. The techno-economic analysis of the obtained results has to be done to select optimal medium composition for industrial production of antimicrobial compounds.

The pollution caused by the use of fossil fuels for the production of mechanical or electrical energy is one of the most important environmental issues nowa­days. In this respect, biofuels represent a viable source of energy. Bioethanol as a renewable energy source is derived from organic material of plant origin, so-called biomass, thus reducing environmental pollution. The aim of this study was to analyze the potential of bioethanol in meeting future energy demands in the Republic of Serbia. [Projekat Ministarstva nauke Republike SRbije, br. TR31002: The improvement of bioethanol production from sugar beet processing products]

In this paper the bioethanol production in batch culture by free Saccharomyces cerevisiae cells from thick juice as intermediate product of sugar beet processing was examined. The obtained results suggest that it is possible to decrease fermentation time for the cultivation medium based on thick juice with starting sugar content of 5-15 g kg-1. For the fermentation of cultivation medium based on thick juice with starting sugar content of 20 and 25 g kg-1 significant increase in ethanol content was attained during the whole fermentation process, resulting in 12.51 and 10.95 dm3 m-3 ethanol contents after 48 h, respectively. Other goals of this work were to investigate the possibilities for experimental results prediction using artificial neural networks (ANNs) and to find its optimal topology. A feed-forward back-propagation artificial neural network was used to test the hypothesis. As input variables fermentation time and starting sugar content were used. Neural networks had one output value, ethanol content, yeast cell number or sugar content. There was one hidden layer and the optimal number of neurons was found to be nine for all selected network outputs. In this study transfer function was tansig and the selected learning rule was Levenberg-Marquardt. Results suggest that artificial neural networks are good prediction tool for selected network outputs. It was found that experimental results are in very good agreement with computed ones. The coefficient of determination (the R-squared) was found to be 0.9997, 0.9997 and 0.9999 for ethanol content, yeast cell number and sugar content, respectively.

Methods that can provide adequate accuracy in the estimation of variables from incomplete information are desirable for the prediction of fermentation processes. A feed-forward back-propagation artificial neural network was used for modelling of thick juice fermentation. Fermentation time and starting sugar content were usedas input variables, i.e. nodes. Neural network had one output node (ethanol content, yeast cell number or sugar content). The hidden layer had nine neurons. Garson's algorithm and connection weights were used for interpreting neural network. The inadequacy of Garson's algorithm can be seen by comparing with the results of regression analysis, which indicates that the influence of the fermentation time is higher. A better agreement of the results was obtained using network connection weights, a method that can be used to determine the relative importance of input variables.

The present study examines xanthan production by Xanthomonas campestris under aerobic conditions on spent grains liquor based cultivation media containing four different amounts of maltose. All four experiments were performed simultaneously, so that all stages of the biotechnological process would be carried out under identical conditions. In addition to the cultivation parameters (pH value, contents of dissolved oxygen, reducing sugars and assimilable nitrogen, turbidity and trickling time), xanthan yield and sugar conversion percentage were also determined. Based on the results of the experiments maltose addition up to 5% to spent grains liquor does not hinder the cultivation course. On the other hand, the conversion of sugars into xanthan decreases with the increase of maltose content to 5% and a large amount of sugars lags in the cultivation broth.

Investigations include the analysis of the impact of commercial products: complex additive (0.1, 0.3, and 0.5%), L-ascorbic acid (0.002, 0.004 and 0.012%), diacetyl ester of tartaric acid with monoglycerides (DATEM E472e, 0.1, 0.3, and 0.5%), α-amylase (0.002, 0.006 and 0.012%), xylanase (0.004, 0.012 and 0.024%), alcohol extract of rosemary, thyme or sage (0.5, 1.0 and 2.0%), as well as the combination of complex additive and rosemary, thyme and sage extract on rheological characteristics of dough. The study includes amylograph, farinograph and extensograph analysis of dough with and without additives (control sample). The volume of lost CO2 gas (mL) is the lowest in dough samples with an added combination of complex additive and thyme extract (0.05 and 0.5%) and rosemary extract (2.0%). In the samples with thyme extract (1.0%) added, the volume of lost gas is at a level of samples with added complex additive, DATEM, and L-ascorbic acid.

The need to freeze dough is increasing in bakery production. Frozen dough can be stored for a long time without quality change. The capacity of bakery production can be increased in this way, and in the same time, the night shifts can be decreased. Yeast cells can be damaged by freezing process resulting in poor technological quality of dough after defrostation (longer fermentation of dough). The influence of frozen storage time of dough on survival percentage of Saccharomyces cerevisiae was investigated. Dough samples were taken after 1, 7, 14 and 28 days of frozen storage at -20°C. After defrosting, at room temperature, samples were taken from the surface and the middle part of dough (under aseptic conditions), and the percentage of living S. cerevisiae cells was determined. During frozen storage of dough, the number of living S. cerevisiae decreased. After 28 days of frozen storage, the percentage of live cells on the surface and inside the dough was 53,1% and 54,95%, respectively. The addition of k-carragenan to dough increased the percentage of living cells in the middle part of dough up to 64,63%. Pure cultures, isolated from survived S. cerevisia cells in frozen dough by agar plates method (Koch's method), were multiplied in optimal liquid medium for yeasts. The content of cytochromes in S. cerevisiae cells was determined by spectrophotometric method. The obtained results showed that the content of cytochromes in survived S. cerevisiae cells was not affected by dough freezing process. Growth rate and fermentative activity (Einchor's method) were determined in multiplied cells.

Processing parameters of Kantata wheat variety indicate that it is not suitable for bread making. Therefore, an investigation was carried out to evaluate the suitability of the variety for ethanol production. Wheat was cultivated at the following sites: Kovin, Zrenjanin, Pančevo and Vrbas. Ethanol yields depend on the location on which the samples were grown and on the temperature during thermal and enzymatic preparation of flour samples. Wheat sample cultivated on the Vrbas site gave the highest ethanol yield.

Dough samples with different content of amylases were investigated immediately after mixing and after 7, 14 and 30 days of frozen storage. The obtained results show that the fermentation time is shorter, both in fresh and frozen samples, when amylase sample 1 was added, compared to dough without enzymes. The addition of amylase 2 to dough resulted in minimal decrease of "rising" time, both is frozen and fresh dough samples. The rising time of fresh samples was shorter when amylase 3 was added to dough. The specific fermentative activity of fresh dough samples is increasing by about 10% compared to the control sample, for all amounts of amylase 1 and 2 added to the do- ugh. The fermentative activity of yeast in frozen samples increased by 5-10%, after keeping of dough with the addition of amylase 1 for 14 days. The specific fermentative activity of fresh dough samples increased compared to the control, for all amounts of added amylase 3 to the dough. In frozen dough samples the fermentative activity of yeast decreased by 10% for all added amounts of amylase 3. Baked goods made of fresh and frozen dough, prepared with the addition of amylase 1, are better than the ones made of control dough sample, considering all evaluated parameters.

The highest amount of zinc (= 90%) is bound after 3 hrs of contact at low initial (total) concentrations of zinc in suspension of yeast, 10-100 mg/l at 10-30°C. The equilibrium between bound and free zinc ions is established after 6 hrs of contact time, independently on the total zinc concentration in yeast milk. No bigger changes of content of zinc bound to brewer's yeast cells was determined at temperatures 10°C and 30°C. 40% of bound zinc in the equilibrium state is bound during the first 15 min of contact of zinc ions and brewer's yeast cells at all initial (total) zinc concentrations in suspension of yeast both at 10°C and 30°C. The "KEKAM" equation can be used for the description of kinetics of zinc biosorption by waste brewer's yeast cells, for the ranges of zinc concentration 10-100 mg/l at 30°C (mean correlation coefficient 0,96) and 60,0-100 mg/l at 10°C (mean correlation coefficient 0,95).

The waste brewer's yeast S. cerevisiae (activated and non-activated) was compared with the commercial baker's yeast regarding the volume of developed gas in dough, volume and freshness stability of produced bread. The activation of waste brewer's yeast resulted in the increased volume of developed gas in dough by 100% compared to non-activated brewer's yeast, and the obtained bread is of more stable freshness compared to bread produced with baker's yeast. The activation of BY affects positively the quality of produced bread regarding bread volume. The volume of developed gas in dough prepared with the use of non-activated BY was not sufficient, therefore, it should not be used as fermentation agent, but only as an additive in bread production process for bread freshness preservation. Intense mixing of dough results in more compressible crumb 48 hrs after baking compared to high-speed mixing.

The evidence of compounds that increase the nutritive value of red wines has been presented in a number of papers. These compounds include catechins and proanthocyanidins among others. Their protective effect on the cardiovascular system and anticarcinogenic properties has been proved. The effect of maceration conditions and increased solid grape parts content, seed in the first place, on the content of phenolic compounds of wine was investigated. Several micro-trials were performed with Cabernet sauvignon sort; in some variants time and temperature of maceration were varied, while the ratio of residual stem and content of seed was increased several times, resulting in a significant change of phenolic compounds content in the obtained wine samples. The presence of ripe stem yielded good results, but only during six days of maceration, while in the case of longer maceration, the change of colour quality was negative. Supplementary quantities of seeds during maceration resulted in an increase of total phenols and catechins. A significant influence on colour of wines was also observed, especially in wines obtained applying shorter maceration.