Rita Scherbaka

Levan-ethanol biosynthesis using Zymomonas mobilis cells immobilized by attachment and entrapment

Fermentation of sucrose by Zymomonas mobilis cells attached to stainless steel wire spheres (WS) and to Al2O3 granules was compared with sucrose fermentation by cells entrapped in Ca-alginate. Similar amounts of cell biomass were applied at the beginning of the immobilized fermentation systems. The immobilization of Z. mobilis cells to the carrier surface was checked by scanning electron microscopy (SEM). Most cells were present in holes and pores of Al2O3 surfaces following colonization. Observation of the carrier after repeated fermentation cycles showed that a surface of Al2O3 granules was partly covered by levan. Alginate beads were extended in volume and partly disrupted. Changing the medium every 48 h established that free Z. mobilis cells in the culture liquid exceeded many times of cells attached to the carrier or incorporated in it. Ethanol and levan production did not differ significantly in these fermentation systems — maximal ethanol concentration at the end of second and third fermentation cycles reached 2.3–2.7% and levan 1.3–1.5%. Fermentation resulted in destruction of Ca-alginate beads. The conclusion was reached, that WS and Al2O3 with immobilized bacteria can be used as an inoculum generator for sucrose fermentation but that the general producer of levan and ethanol is the free suspended cell biomass.

Carbohydrates in Jerusalem artichoke powder suspension

Purpose – The aim of this study was to evaluate the influence of temperature and action time on the extraction rate of carbohydrates of Jerusalem artichoke concentrate powder and inactivation of inulin during boiling and sterilization.Design/methodology/approach – Water suspension of Jerusalem artichoke concentrate (5g/100ml) at 25, 50 and 100○C was tested after 5, 15, 30 and 60min to determine the content of inulin, glucose, fructose and sucrose and evaluate the extraction rate. The stability of inulin was studied after boiling and sterilization at 120○C during 1, 2 and 3h. The extraction rate was evaluated by Fourier‐Transform Infrared (FT‐IR) spectroscopy as well.Findings – It was shown that extraction of soluble carbohydrates – fructose, glucose, sucrose and inulin, from water suspension of Jerusalem artichoke concentrate was practically completed in 5min at 25○C. The extraction rate was not significantly influenced by temperatures lower than 100○C. Inulin was stable during boiling for 1h but steriliz...

Effects of yeast immobilization on bioethanol production

The current study evaluated a newer method, which includes a dehydration step, of immobilizing Saccharomyces cerevisiae L‐77 and S. cerevisiae L‐73 onto hydroxylapatite and chamotte ceramic supports. The efficiency of cell immobilization on chamotte was significantly higher than hydroxylapatite. Immobilized yeast preparations were investigated for their ethanol‐producing capabilities. The glucose concentration in a fermentation medium was 100 mg/mL. Immobilized preparations produced the same amount of ethanol (48 ± 0.5 mg/mL) as free cells after 36 H of fermentation. During the early stages of fermentation, immobilized yeast cells produced ethanol at a higher rate than free cells. Yeast preparations immobilized on both supports (hydroxylapatite and chamotte) were successfully used in six sequential batch fermentations without any loss of activity. The chamotte support was more stable in the fermentation medium during these six cycles of ethanol production. In addition to the high level of ethanol produced by cells immobilized on chamotte, the stability of this support and its low cost make it a promising material for biotechnologies associated with ethanol production.

Effect of plant extract on the degradation of nitroaromatic compounds by soil microorganisms

Remediation of soils contaminated by nitroaromatic compounds and nitramines, i.e. explosives, is known as very important, complicated, and rapidly developing area of biotechnology. A search for optimal growth conditions for soil bacteria is of a great importance in order to isolate various xenobiotic degraders. Bacteria consortium A43 was isolated from soils contaminated with explosives. In the presence of carbohydrate and plant extract, an addition of TNT to the solidified minimal medium stimulated the growth of the tested bacteria, as compared to other bacteria consortium isolated from the same soils. Reducing sugars as carbohydrates, and cabbage leaf extract as a plant extract were used in these experiments. Cultivation of the A43 in liquid medium of the same content showed that addition of cabbage leaf extract alone to medium is much more efficient for TNT degradation by growing biomass as compared to addition of carbohydrate alone.

Sugar beet diffusion juice and syrup as media for ethanol and levan production by Zymomonas mobilis

Abstract Sugar beet is an important crop cultivated widely in Europe. Roots contain up to 6–9 t/ha of fermentable sugar. Our laboratory is targeting to investigate the production possibilities of ethanol and levan by Zymomonas mobilis bacteria, using sugar beet juice or their syrup as raw material. The Z.mobilis strain 113 “S”;, selected in our institute, produces simultaneously with ethanol up to 40–70 g/1 fructose polymer levan in sucrose medium. It was established that Z.mobilis can convert sucrose into ethanol and levan in batch fermentation using natural sugar beet diffusion juice, without additives of mineral salts and growth factors. Syrup obtained in sugar industrial production by conventional technology from diffusion juice after evaporation with 65% sucrose can be used also as raw material for medium preparation. Additives of N, P, K, Mg, as well as yeast extract, were shown to stimulate the culture growth and product synthesis on such media. Reuse of liquid fermentation products after separatio...

Fermentation of Jerusalem artichoke by Zymomonas and Saccharomyces

Purpose – The aim of this study is to find out the appropriate fermentation conditions of Jerusalem artichoke powder (JAP) based media to obtain light beverage containing inulin.Design/methodology/approach – JAP water suspension or filtrate were used for preparation of growth media with or without enzymatic hydrolysis of inulin for fermentation by Zymomonas mobilis 113 “S” or Saccharomyces cerevisiae.Findings – If enzymatic hydrolysis of inulin was not used significantly higher amount of inulin (7.42 per cent) was unconverted by Z. mobilis than by S. cerevisiae (2.22 per cent) while the ethanol concentration was much higher (2.86 per cent) after S. cerevisiae fermentation than after Z. mobilis fermentation (1.21 per cent). Considerably more ethanol was produced by Z. mobilis during co‐fermentation with Fructozyme L of JAP suspension filtrate (5.98 per cent) and suspension (4.96 per cent). Analyses of volatile components of fermentation broths showed that for production of inulin containing light beverages...

Model-based biotechnological potential analysis of Kluyveromyces marxianus central metabolism

The non-conventional yeast Kluyveromyces marxianus is an emerging industrial producer for many biotechnological processes. Here, we show the application of a biomass-linked stoichiometric model of central metabolism that is experimentally validated, and mass and charge balanced for assessing the carbon conversion efficiency of wild type and modified K. marxianus. Pairs of substrates (lactose, glucose, inulin, xylose) and products (ethanol, acetate, lactate, glycerol, ethyl acetate, succinate, glutamate, phenylethanol and phenylalanine) are examined by various modelling and optimisation methods. Our model reveals the organism’s potential for industrial application and metabolic engineering. Modelling results imply that the aeration regime can be used as a tool to optimise product yield and flux distribution in K. marxianus. Also rebalancing NADH and NADPH utilisation can be used to improve the efficiency of substrate conversion. Xylose is identified as a biotechnologically promising substrate for K. marxianus.