Dragiša S. Savić

PROGRESS IN THE PRODUCTION OF BIOETHANOL ON STARCH-BASED FEEDSTOCKS*

Bioethanol produced from renewable biomass, such as sugar, starch, or lignocellulosic materials, is one of the alternative energy resources, which is both renewable and environmentally friendly. Although, the priority in global future ethanol production is put on lignocellulosic processing, which is considered as one of the most promising second-generation biofuel technologies, the utilization of lignocellulosic material for fuel ethanol is still under improvement. Sugar-based (molasses, sugar cane, sugar beet) and starch-based (corn, wheat, triticale, potato, rice, etc.) feedstock are still currently predominant at the industrial level and they are, so far, economically favorable compared to lingocelluloses. Currently, approx. 80 % of total world ethanol production is obtained from the fermentation of simple sugars by yeast. In Serbia, one of the most suitable and available agricultural raw material for the industrial ethanol production are cereals such as corn, wheat and triticale. In addition, surpluses of this feedstock are being produced in our country constantly. In this paper, a brief review of the state of the art in bioethanol production and biomass availability is given, pointing out the progress possibilities on starch-based production. The progress possibilities are discussed in the domain of feedstock choice and pretreatment, optimization of fermentation, process integration and utilization of the process byproducts.

A survey of the lactic acid bacteria isolated from Serbian artisanal dairy product kajmak

Kajmak is an artisanal Serbian dairy product made by fermentation of milk fat. Overall, 374 bacterial isolates were collected from six kajmak samples of different ages produced in the households located in distinct regions of Serbia. In order to identify lactic acid bacteria present in chosen samples of kajmak, total 349 Gram-positive and catalase-negative isolates were analyzed. The recognition of isolates was performed by phenotypic characterization followed by molecular identification using (GTG)(5)-PCR and sequence analysis of 16S rRNA gene. Leuconostoc mesenteroides and Enterococcus faecium were the most frequently isolated species from kajmak samples. In contrast, leuconostocs and enterococci were found in BGMK3 and BGMK1 kajmak respectively, only after using enrichment technique for isolation suggesting they are present in low numbers in these kajmaks. Lactococcus lactis, Lactococcus raffinolactis and Lactococcus garvieae were also found in those samples but in lower proportion. Results showed that Lactobacillus plantarum, Lb. paracasei and Lb. kefiri were the most frequently isolated Lactobacillus species in analyzed kajmaks.

The characterisation of lactic acid bacteria during the fermentation of an artisan Serbian sausage (Petrovská Klobása)

Petrovská Klobása is an artisan Serbian sausage made only from meat and spices without any additives or starter cultures. In order to characterise lactic acid bacteria (LAB) microflora, a total number of 404 LAB strains were isolated from 15 samples collected during 90 days of the fermentation and 120 days of storage of one batch of Petrovská Klobása. The isolates were preliminarily identified by phenotypic tests and subjected to (GTG)₅-PCR fingerprinting. Representatives of each group were identified by 16S rDNA sequencing. The results showed that among the isolates, Lactobacillus sakei and Leuconostoc mesenteroides predominate with 36.4% and 37.1% of total LAB strains, respectively. Pediococcus pentosaceus was also isolated in high proportion (18.4%) whereas Enterococcus durans and Enterococcus caseliflavus made only 1% and 6% of the total isolates, correspondingly. The analysis of vacuum packed and modified atmosphere packed (MAP) samples showed higher presence of L. mesenteroides and L. sakei in the total microflora.

Effects of aeration intensity on pyrite oxidation by Thiobacillus ferrooxidans

The oxidation of pyrite in suspension by the bacterium Thiobacillus ferrooxidans B5 was studied under different aeration conditions in the shake flasks and a bubble column to establish the effects of aeration intensity on the bioprocess. Agitation is of the greatest importance for the bioprocess and has to be adequately intensive to suspend pyrite particle in the medium. Stronger agitation decreases biodissolution efficiency, due to the reduced action of the direct mechanism in the first part of the bioprocess. Although biooxidation in the bulk solution depends on ferrous iron realized from pyrite by direct bacterial actions, a stronger intensity reduces the contribution of the free suspended cells in the biooxidation process.

Oxygen mass transfer requirements during ferrous iron oxidation by Thiobacillus ferrooxidans under controlled pH conditions

The ferrous iron oxidation by the bacterium Thiobacillus ferrooxidans B5 was studied under different aeration conditions at constant pH (1.8 and 2.0) in three types of bioreactors (shake flasks, a bubble column and a stirred tank). The main goal was to reflect the effects of the method and the intensity of agitation and/or aeration on ferrous iron biooxidation. The specific growth rates and the substrate oxidation rate constants in the exponential phase were actually the same (0.069 ± 0.002 h-1), regardless of the bioreactor type and the pH condition, if the oxygen transfer rate was higher than approximately 1.0 mg O2/L/min. The maximum biooxidation constants were affected by the oxygen transfer only at lower aeration intensities. Thus, the oxygen transfer rate could be used as a basis for scaling-up bioreactors for the ferrous iron biooxidation by T. ferrooxidans under aerobic and controlled pH condition.

Production of biodiesel from microalgae

In recent years, more attention has been paid to the use of third generation feedstocs for the production of biodiesel. One of the most promising sources of oil for biodiesel production are microalgae. They are unicellular or colonial photosynthetic organisms, with permanently increasing industrial application in the production of not only chemicals and nutritional supplements but also biodiesel. Biodiesel productivity per hectare of cultivation area can be up to 100 times higher for microalgae than for oil crops. Also, microalgae can grow in a variety of environments that are often unsuitable for agricultural purposes. Microalgae oil content varies in different species and can reach up to 77% of dry biomass, while the oil productivity by the phototrophic cultivation of microalgae is up to 122 mg/l/d. Variations of the growth conditions and the implementation of the genetic engineering can induce the changes in the composition and productivity of microalgal oil. Biodiesel from microalgae can be produced in two ways: by transesterification of oil extracted from biomass or by direct transesterification of algal biomass (so called in situ transesterification). This paper reviews the curent status of microalgae used for the production of biodiesel including their isolation, cultivation, harvesting and conversion to biodiesel. Because of high oil productivity, microalgae will play a significant role in future biodiesel production. The advantages of using microalgae as a source for biofuel production are increased efficiency and reduced cost of production. Also, microalgae do not require a lot of space for growing and do not have a negative impact on the global food and water supplies. Disadvantages of using microalgae are more difficult separation of biomass and the need for further research to develop standardized methods for microalgae cultivation and biodiesel production. Currently, microalgae are not yet sustainable option for the commercial production of biodiesel. First of all, the price of biodiesel from microalgae is still higher than the price of diesel due to high production costs. [Projekat Ministarstva nauke Republike Srbije, br. III 45001]

Determination of CO2 Content in the Headspace of Spoiled Yogurt Packages

The CO2 formation during food storage can often be correlated with the increase in yeast population. Yogurt and other dairy products are susceptible to yeast contamination. Accumulation of CO2 in the headspace of yogurt packages can lead to the eventual blowing off of the package. Therefore, determination of CO2 in the yogurt packages can indicate eventual unsafety of the product. The aim of this paper was to determine CO2 concentration in the headspace of different yogurt containers contaminated with yeast at the levels of 1 and 5 CFU/ml. Yeast Candida kefyr, previously isolated from spoiled yogurt, was used for contamination. Contaminated and control samples of yogurt were incubated at 30°C. A device based on tunable diode laser absorption spectroscopy was used for the measurement of CO2 concentration. The CO2 content in all analysed samples changed in a similar manner with slow increase to the value of 6% during the first 30 h and, after that, rapid accumulation to 17–20%. The initial level of yeast contamination did not have significant influence to the CO2 content trend. The increase in the number of yeast was observed after 10 h of incubation, and the final value of 6-7 log·CFU/cm3 was reached after 40 h of incubation. The significant increase in the yeast number can be correlated with the CO2 content in a way that CO2 concentration of 6% can be considered as critical for microbial spoilage. Since the TDLAS technique is simple and nondestructive, it can be a promising possibility for detection of the microbial spoilage in food.

The isolation and screening of microalgae for the production of oil

The biodiesel production has gained increasing attention of the researches in recent years. Current commercial biodiesel production involves transesterification of oil derived from oil crops. Since this production is no more sustainable, the use of microalgae represents a good alternative. Microalgae have high growth rate, high oil content and can be cultured in the environment which are not suitable for agriculture. Additionally, microalgae cultivation improves the reduction of carbon dioxide in the atmosphere. Selection of microalgae for oil production must take into consider certain criteria which include growth rate, oil content, fatty acids profile and ease of separation. In order to analyze the possibility of the use of microalgae for the production of oil, isolation of freshwater microalgae was performed. The isolation was done by the use of traditional techniques from freshwaters near Leskovac. A total number of 6 microalgae strains were isolated and identified as the representatives of the genera Chlorococcum (1 isolate), Chlorella (1 isolate), Scenedesmus (1 isolate) and Desmodesmus (3 isolates). Isolates of microalgae were screened for the growth rate, biomass and oil productivity and oil content. The highest content of biomass was 1.5 g/l and it was observed in the strains Chlorococcum sp. and Desmodesmus sp. 1. The highest value of specific growth rate was calculated during the growth of microalgae Chlorella sp., Scenedesmus sp. and Desmodesmus sp.1, while the lowest value was observed for Chlorococcum sp. Oil productivity was the highest for Chlorella sp. and Desmodesmus sp.1 (0,4 g/l) and the lowest for Desmodesmus sp.2, Desmodesmus sp.3 and Chlorococcum sp. (0,2 g/l). The oil content was in the range from 15.8% (Chlorococcum sp.) to 33% (Chlorella sp). Since the microalgae isolates Chlorella sp. and Scenedesmus sp. had the highest oil productivity, high growth rate and high oil content, these strains are the most suitable for further investigation in order to improve the oil yield and analyse the possibility of the use in the production of biodiesel.

The Influence of Modified Media with Carboxymethyl Cellulose on Morphology and Antibiotic Production by Streptomyces Hygroscopicus

ABSTRACT Actinomycetes, especially members of the genus Streptomyces, are responsible for producing the majority of known antibiotics. The production of antibiotics by filamentous organisms is often dependent on the morphology and size distribution of the pellet population within the culture. The effect of carboxymethylcellulose (CMC) on growth and antibiotic (hexaene and azalomycine) production by Strepotmyces hygroscopicus were investigated. By adding the polymer to the fermentation medium, the growth was changed from a single large glob to small reproducible pellets, and wall growth was diminished to a minimum. Maximum concentration of hexaene was reached at 3% (wt/vol) CMC and maximum concentration of azalomycine was reached at 1 % (wt/vol) CMC.