Library

Notes: In situ hybridization with fluorescently monolabelled rRNA-targeted oligonucleotide probes (17 to 18 nucleotides) was used to discriminate between Alcaligenes eutrophus JMP 134 and Acinetobacter calcoaceticus 69-V by flow cytometry. The strains were grown in batch experiments in a mixed population. The forward light scatter and fluorescence of each bacterial cell were measured with a single laser cytometer. The intensity of fluorescence after rRNA staining depended on the content of ribosomes, which correlated with the growth rate of bacteria. Therefore exponentially growing cells could be clearly detected. For other growth phases, signal amplification was necessary using multiple probes. The two bacterial strains were identified with differently labelled probes under an epifluorescent microscope. Using a single laser cytometer, rRNA based identification was possible nut not ideal. Better discrimination between the two strains of the mixed population was achieved by DNA staining, combined with the different forward light scatter signals. Due to the significantly different cellular DNA and GC content of both strains, the fluorescent dye DAPI (4′, 6-diamidino-2-phenylindole), preferring AT-rich regions of DNA, was found to be a supplementary tool for population analysis. The abundance ratios of the two strains in mixed culture determined by DNA or rRNA staining were similar.

Notes: The process of leachate denitrification by populations of nitrifying and denitrifying bacteria was investigated. Leachate, derived from a local municipal landfill site, was nitrified in a continuously operating packed-bed biofilm reactor and thereafter denitrified in an activated sludge bioreactor. To follow the progress of nitrogen elimination, ammonium, nitrite and nitrate concentrations were determined at all stages of the process. While the nitrite and nitrate concentrations were measured by conventional colorimetric methods, computer controlled coulometric titration with in situ generated hypobromite was used for ammonium determination, which had previously been selectively separated from the sample matrix by gas dialysis. The detection range of the method was from 1 × 10-6 to 1 × 10-3 M ammonium (relative standard deviation (RSD) = 2%, n = 6). No interference of the complex sample matrix was found in ammonium determination. The average ammonium concentration in the leachate was 409 mg/l (standard deviation (SD) = 142 mg/l, n = 55). The ammonium concentrations decreased to 1-5 mg/l during nitrification under continuous operating conditions. Increased ammonium concentrations after nitrification correlated with a decrease in the efficiency of nitrogen elimination by up to 45% due to the build-up of high concentrations of nitrite. The concentration of sulphides, another source of pollution of the leachate, was also determined by triangle programmed coulometric titration. The average concentration of sulphides in the leachate was 221 mg/l (SD = 374 mg/l; n = 55). The sulphide concentrations decreased to concentrations below the detection limit of the coulometric titration (2 × 10-6M) during nitrification.

Notes: An enzymatic membrane for application in the processes of decomposition and removal of urea from aqueous solutions was prepared: jack bean urease was immobilized on an aminated polysulphone membrane by adsorption. The inhibition of the system by boric acid was studied using procedures based on the MICHAELIS-MENTEN integrated equation (non-linear regression, and the linear transformations of WALKER and SCHMIDT, JENNINGS and NIEMANN, and BOOMAN and NIEMANN). The reaction was carried out in a 100 mM phosphate buffer of pH 7.0, containing 2 mM EDTA, obtained by neutralization of orthophosphoric acid with NaOH, at an initial urea concentration of 10 mM, and a temperature of 25 °C. The reaction was initiated by the addition of the enzyme to the urea solution, and was monitored by removing samples of the reaction mixture for NH3 determinations by the phenol-hypochlorite method until the urea was exhausted. The results were compared with those obtained earlier under the same reaction conditions for free urease and urease covalently immobilized on chitosan. The inhibition was found to be competitive, similar to that of the free enzyme and urease immobilized on chitosan, with inhibition constants Ki equal to 0.36, 0.19 and 0.60 mM. The results show that adsorption of the enzyme on a polysulphone membrane changed the enzyme to a lesser degree than covalent immobilization of the enzyme on a chitosan membrane.

Notes: Investigations were conducted with the aim of producing extracellular lipase from Candida rugosa by solid-state fermentation (SSF), using coconut oil cake (COC) as a solid substrate. To optimize production, various modifications were made to enrich the substrate by supplementing it with mineral solution, different carbon sources and several inorganic as well as organic nitrogen sources. Among them, urea (1%), peptone (3%) and maltose (5%) were found to be most suitable. Addition of olive oil (10%) encouraged lipase synthesis. The maximum lipase activity in the enriched substrate was 87.76 units per gram of dry fermented substrate [U/gds] compared to 25.81 U/gds in the raw cake at 96 h of fermentation, and growth was as high as 14.44 mg/gds of glucosamine. This was reached at 72 h in the enriched substrate. C. rugosa growth was calculated indirectly by estimating the glucosamine content in the cell wall after its hydrolysis. The enzyme yield was far better than any values reported as yet.

Notes: In this work, an immobilization method for polymer-levan production by a non-flocculating Z mobilis culture was developed. The extent of cell attachment to the stainless steel wire surface, culture growth and product synthesis were described. It was established that during short-term passive immobilization of non-flocculation Z mobilis cells on a stainless steel wire surface, sufficient amounts of biomass for proper levan and ethano fermentation could not be obtained. Adherence of cells was improved by pressing the paste-like biomass within stainless steel spheres knitted from wire with subsequent dehydration. Biomass fixed in metal spheres was used for repeated batch fermentation of levan. The activation period of cells within wire spheres (WS) was 48 h in duration. During this time, cell growth stabilized at production levels of ethanol and levan of Qeth = 1.238 g/l × h and qeth = 0.47 g/l × h; Qeth = 0.526 g/l × h and qeth = 0.20 g/l × h. Five stable fermentation cycles were realized using one wire sphere inoculum, and maintaining a stable ratio of 2.4 of biomass suspended in the medium to biomass fixed in the sphere. Using fixed Z mobilis biomass in the WS, the total amount of inoculum could be reduced for batch fermentation. Large plaited wire spheres with biomass may have potential in fermentation in viscous systems, including levan production.

Notes: This review is concerned with the application of hairy roots, i.e. plant roots formed from plant cells after transformation by Agrobacterium rhizogenes for the production of bioactive compounds. Transformed root cultures have been established from numerous species of dicotyledonous plants. The plants, as well as the main products accumulated in hairy root cultures derived from these plants, are listed in this paper. Data are presented on novel compounds, hitherto detected only in transformed roots but not occurring in the corresponding intact plants.The possible use of hairy root cultures for the over-production of secondary metabolites and biotransformation of chemicals is discussed. In order to enhance the productivity of hairy root cultures, various methods have been derived, and optimized procedures are proposed. They include selection of high-producing clones, elicitation, composition of growth media, culture conditions and genetic approach. Hairy roots usually store secondary metabolites in vacuoles inside the cells. Therefore, several methods have been used to increase the amount of products released into the medium. Unfortunately, no general procedure is known that works in all cases, and the excretion behaviour of hairy root cultures varies from one species to another and even within one species from one clone to another.Special attention is given to the cultivation methods and bioreactor systems for hairy root cultures. Hairy roots are cultivated usually in shake flasks; however, shake flask culture is not suitable for the complex optimization and continuous control of the culture conditions. In this paper, we are going to present bioreactors proposed for the cultivation of hairy roots under more or less controlled conditions. Modifications of typical bacterial bioreactors, i.e. stirred tanks, airlift loop reactors and other constructions, are presented. A very special type of bioreactor providing good conditions for loose root mass multiplication without oxygen or substrate limitations, is the mist bioreactor. Nowadays, it is practically impossible to select the one best bioreactor type for hairy root culture.

Notes: Hydrolyses of olive oil were performed in a reactor with lipase immobilized on a laboratory ultrafiltration poliamide-6 membrane. The reactor consisted of two circulating phases of olive oil and buffer solution. For the characterization of the reactor performance, a model of the hydrolysis process was developed. It was created by means of thermodynamic network representation of both the chemical processes and the transport of the reactants. According to an estimated bond graph network, the model is represented quantatively by a set of thirty-three differential equations representing the time derivatives of the particular species concentration. The parameters of the model were estimated based on experimental data and/or literature notations. Close agreement of numerical estimations of the product concentrations with experimental data was gained. The model enabled an extended analysis of the influence of different reaction parameters, enzyme inhibition and concentration of the reactants on reactor performance.

Notes: The effects of anoxic conditions on product inhibition and the stability of L-ATC hydrolase were investigated in the conversion of D,L-2-amino-Δ2-thiazoline-4-carboxylic acid (D,L-ATC) to L-cystine using the cell free extract enzyme of Pseudomonas sp. in the presence of hydroxylamine. At L-cysteine equivalent levels, where one mole of L-cystine was counted as two moles of L-cysteine, L-cystine inhibited the L-ATC hydrolase reaction to a greater extent than L-cysteine. In air, the product occurred predominantly as L-cystine (94.9%), whereas in a nitrogen atmosphere the product occured as a mixture of L-cysteine (39.3%) and L-cystine (40.7%). As a result, less product inhibition took place in nitrogen. The activity of L-ATC hydrolase was almost fully lost after 20 h of incubation by shaking at 30 °C in air, but considerable activity remained under the anoxic conditions of nitrogen. A kinetic analysis of the reactions confirmed that reduced product inhibition and enhanced enzyme stability in nitrogen result in a more efficient enzyme reaction. The inactivation rate constant (k1) was estimated to be 0.11 h-1 in nitrogen and 0.22-1 in air, indicating that the stability of L-ATC hydrolase in nitrogen was greater than in air. The values of the Kp1 and Kp2 constants related to product inhibition were 43.36 mM and 30.48 mM for L-cysteine and L-cystine, respectively, where higher values were an indication of less product inhibition. The value of the rate constant (k2) for the oxidation of L-cysteine to L-cystine was 0.09 h-1 in nitrogen and 1.01 h-1 in air, suggesting that the oxidation of L-cysteine to L-cystine proceeds faster in air than in nitrogen.

Notes: Enzymatic stereoselective hydrolysis of (R,S)-1-phenylethyl propionate was performed in a stirred tank and in a biphasic enzyme membrane reactor. Lipase from Pseudomonas sp. was proved to be a good enantioselective catalyst for this reaction. The enzyme was covalently immobilized in a porous polyamide membrane (flat sheet as well as hollow-fibres) via glutaraldehyde. An influence of membrane hydrophobicity on reactor performance was observed. Initial lipase activity and productivity in the processes were equal to 1.05 × 10-4, 1.3 × 10-5 and 1.0 × 10-5 mole/(h × mg of enzyme) in the case of native lipase, in the aromatic polyamide hydrophobic membrane reactor and in the hydrophilic polyamide-6 membrane reactor, respectively. The influence of some factors such as temperature, pH, buffer concentration, initial substrate concentration and addition of β-cyclodextrin derivatives on reaction rate and enantioselectivity was investigated and discussed. In the enzyme membrane reactor both organic and aqueous phases circulated countercurrently on both sides of the membrane. At a conversion degree of under 55-60%, pure enantiomer of the remaining ester (i.e. 〉 98%) was obtained.

Notes: A real-time artificial intelligence method for controlling the concentration of dissolved oxygen is proposed.Two projection versions of algorithms are considered in this paper. The versions vary in structure of the intelligent agents. One of the agents represents an automaton of expedient behaviour, the structure of the other consists of two automata which behave expediently in a complicated random medium.The first algorithm holds a check on a trend of change in values. The second version of the algorithm takes into account both the trend of change in the values and the speed of change in the values.Simulation studies show that expedient behaviour of the automata in the random medium for the control of dissolved oxygen concentration can bring about a good performance.

Notes: The production of L-glutamic acid with Corynebacterium glutamicum under biotin limitation was studied. Assuming a formal type of cell maturation, an adequate formal kinetic model was developed. This model includes growth, dependent on biotin, and uses the same retention term for describing the lag phase and cell maturation. Special attention was paid to the graphical interpretation of the performance between the variables, which is relevant for kinetics. Comparison between experiments and the model resulted in different degrees of agreement. However, the main trend of the experimental patterns of the complex bioprocess can clearly be mirrored in this model.

Notes: Bacterial strains have been isolated from contaminated concrete debris which exhibit the metabolic capability to degrade 2,4-chlorinated and 4-chloro-2-methyl-substituted phenols and phenoxyalkanoic acids including phenoxyacetate and phenoxypropionate derivatives. These strains were taxonomically identified. Two of them were found to belong to the β-subgroup of the proteobacteria and showed strong similarity to Rhodoferax fermentans. Preliminary investigations by PCR amplification using respective primers revealed that the strains harbour tfdA-like gene sequences.

Notes: The cell suspension culture Daucus carota L. was permeabilized by Tween 80 and immobilized by glutaraldehyde. β-Galactosidase showed an optimum pH of 4.7 and an optimum temperature of 55 °C. The enzyme hydrolysis was linear for 3 h, reaching a 65% conversion. A very good level of storage stability was achieved when using dry catalyst, or a solution of 0.15 M NaCl with the addition of chloramphenicol, (l-methyldodecyl)-dimethylamin-4-oxide (ATDNO), chlortetracycline hydrochloride (CLCTC) or by freezing the immobilized cells in 0.15 M NaCl. The cells characterized by high enzyme activity and stability in long-term storage showed convenient physicomechanical properties.

Notes: A new finite-state method is proposed which has been designed for use in biotechnological processes, in particular for the control of the pH in acidic waste water.The automation of expedient behaviour takes into account the non-linear character of the process and a good control stability in spite of variations in the influent acidic concentration, dissociation constant of the acid and change of the pH set point.To design the controller with the proposed method, no model of the process is required. Simulation studies show that expedient behaviour of an automaton in a random medium for the control of the pH neutralization process can give a good performance.

Notes: Cells of an Actinomycete-like bacterium, strain GJ70, with the ability to degrade several haloalkanes were used as a biological component in a discontinuous microbial bioassay for the detection of 1,3-dichloropropene and 1,2-dibromoethane in water. The cells were entrapped in different matrices such as calcium alginate, carrageenan, chitosan, polyacrylamide-hydrazide and chitosan-carboxy-methyl cellulose; the specific dehalogenating activity of the immobilized cells to a stirred sample solution and by the use of an ion selective electrode (ISE) for the quantification of enzymatically released halogen ions, the concentration of halogenated hydrocarbons could be estimated by determining the change of electrode potential within a period of 5 min. The detection limits for 1,3-dichloropropene and 1,2-dibromoethane were below 100 μg/l and 25 μg/l, respectively; the relative standard deviation was 〈 10%. In addition, several chlorinated and brominated hgydrocarbons were converted by the bacterial cells at a reduced rate e.g. 1, 2-dibromopropane, 1-bromoethane, 1,5-dichloropentane, etc. Moreover, temperatures of between 20 and 40%C did not affect the enzymatic activity of the cells, and a pH of between at 5 and 9 had little influence. Several organic substances and non-metabolizable compounds did not affect the conversion, whereas some heavy metal ions acted as inhibitors.

Notes: Forty-two white-rot fungi isolated in South America were incubated with long fibre sugar cane bagasse (LFB). The residual composition of LFB was determined after white-rot decay at 30 and 60 days. The ratio of residual lignin to residual lignin to residual cellulose (RL/RC) of untreated material (LFB) was 0.48. After white-rot-decay, the residual material with lower RL/RC ratios indicated that mainly lignin was degraded. In only 30 days, Phlebia sp. MVHC 5535, Athelia sp. MVHC 5509 and Spongipellis pachyodon MVHC 5019 caused a decrease in the RL/RC ratio to 0.36, 0.37 and 0.38, respectively, while it took 60 days for Ganoderma applanatum MVHC 5347, Hyphodontia sp. MVHC 5544, Panus tigrinus MVHC 5400, Stereum sp. MVHC 5113, Phellinus punctatus MVHC 5346 and MVHC 6388 to reach a ratio lower than 0.40. No correlation was found between the amount of some ligninolytic enzymes secreted and the residual composition of bagasse after white-rot fungi fermentation. Most of the fungal strains caused an increase in the relative amount of residual cellulose, indicating that hemicellulose was the preferred energy source.

Notes: A poppy cell suspension culture was permeabilized by Tween 80 and immobilized by glutaraldehyde. The α-Galactosidase in these cells showed an optimum pH level at 5.2 and an optimum temperature at 70 °C. Enzyme hydrolysis was linear for 3 h, reaching 86% conversion. A very good level of storage stability was achieved when using dry catalyst and immobilized cells in 0.15 M NaCl solution (with the addition of chloramphenicol, [1-methyldodecy1)-dimethylamin-4-oxide (ATDNO), chlortetracycline hydrochloride (CLCTC)] or by freezing them in 0.15 M NaCl solution.

Notes: R. opacus UFZ B 408 is able to use pyridine, a potentially growth-inhibiting substrate, as the sole source of carbon, energy and nitrogen. In a previous publication [1] we reported that with the simultaneous utilization of a second carbon and energy source in carbon-substrate-limited chemostat culture, stable steady states could be achieved at higher dilution rates than with growth on pyridine as the sole substrate. Owing to the higher growth yield during growth on such a substrate mixture, both the specific pyridine consumption rates and the residual pyridine concentrations were lower at similar dilution rates than with growth on pyridine alone. Therefore, the critical growth-inhibitory pyridine concentration was only achieved at a higher dilution rate.With the investigations presented here in carbon-substrate-limited continuous culture, the simultaneous utilization of pyridine and formate by R. opacus UFZ B 408 was studied. The yield coefficient during growth on pyridine as the sole substrate amounted to about 0.55 g dry mass/g pyridine. Theoretically, however, the carbon-metabolism-determined yield coefficient should have been about 0.915 g dry mass/g pyridine. Because of the difference between these two values the conclusion was drawn that pyridine is energetically deficient. That means that during growth on pyridine a part of the substrate was dissimilated to supply the energy required for the incorporation of the pyridine carbon into biomass. Formate cannot be used as a carbon source for growth by R. opacus UFZ B 408. However, with growth on pyridine, formate was oxidized simultaneously. During growth on pyridine/formate mixtures, the yield coefficient could be enhanced up to 0.7 g dry mass/g pyridine. That means that biologically usable energy, generated in the course of the formate oxidation, was used for the assimilation of pyridine carbon. The increase in the yield coefficient was related to the utilization ratio of formate to pyridine in a linear manner. However, the carbon-metabolism-determined yield coefficient of 0.915 g dry mass/g pyridine could not be achieved. That can be put down to the fact that R. opacus UFZ B 408 possesses only a limited capacity to oxidize externally supplied formate. Because of the limited formate oxidation capacity the probability is low that, with simultaneous utilization of formate, stable steady states could be achieved at substantially higher dilution rates than with growth on pyridine alone.Enzymatic studies revealed the induction of both NAD(P)+-linked glutaric dialdehyde dehydrogenase and isocitrate lyase during growth on pyridine. Therefore, the conclusion was drawn that pyridine is metabolized by R. opacus UFZ B 408 via the same pathway described for the utilization of pyridine by Nocardia Z1 [2]. This conclusion implies that the ability to oxidize formate represents a metabolic performance which seems not to be directly related to the pyridine metabolism of R. opacus UFZ B 408.

Notes: Hydrolysis and transformation of Fibrenier cellulose (USA) with enzymes from Aspergillus niger IBT-90 was studied. The process was performed at 50°C and pH 4.8 for 24 h using an enzyme complex either as a properly diluted culture filtrate or as a mixture of isolated and purified enzymes from A.niger IBT-90. In the latter experiments, enzyme-substrate ratios expressed as units of activity per 1 g of cellulose were as follows: endoglucanase E1 and E2, 40; β-glucosidase, 40 and cellobio-hydrolase, 2. Cellulose concentration was 5%. It was proved that the crude celluloytic complex from A. niger IBT-90 exhibits higher efficiency in the decomposition of cellulose in comparison to the mixture of enzymes isolated from this complex, as was revealed in assays of reducing sugars and determinations of light transmission throughout cellulose fibres using a computer analysis of the microscopic image. Comparison of both the endoglucanases E1 and E2 showed that the first enzyme is more active against cellulose. It liberated more reducing sugars and caused more significant decomposition of fibres. The predominant effect of the endoglucanase E2 was a smoothing of the fibre surface. The cellobiohydrolase split a cellulose fibre into many short fibres.

Notes: The enzyme Glucoamylase [1,4-α-D-glucan-glucohydrolase EC 3.2.1.3] is very important for the food industry. It is used for producing glucose, ethanol and beer, as well as in technological processes that require the decomposition of starch. Eight mutants of the species Aspergillus niger are evaluated and tested with respect to their production of Glucoamylase and proved to be suitable. The task is to find the mutant showing the highest enzyme activity with a given precision. Conventionally, this kind of multiple decision problem is handled by the analysis of variance (Model I), which tests the homogeneity of the population means, but in this case the results do not supply the desired information. Provided that the enzyme activities of the mutants are different, selection procedures can be used to choose the mutant with the “best” or at least a “good” level of activity.In this paper, a short methodical summary about the two classes of selection procedures is given, i.e. the indifference zone (and d-correct) procedures and the subset procedures. By the example of the selection of a mutant with high enzyme activity the planning of experiments is shown. Depending on suppositions about the variances, different selection rules are applied. Starting with the subset procedure of GUPTA, the number of mutants is reduced to seven. The following application of the d-correct procedures of BECHHOFER, DUNNETT and SOBEL allow us to calculate the necessary sample size of n = 49. Then the mutant whose sample has the largest mean will be selected as a “good” one with a given precision of d = 4 [u/l] and a probability of correct selection of (1-β) = 0.9This application is result of a cooperation between the Dept. of Food of the Technical University, Berlin, and the Dept. of Biotechnology of the Higher Institute of Food and Flavour Industry, Plovdiv, sponsored by the DAAD andthe TU Berlin.

Notes: Three different kinds of biomass, namely Populus deltoides, Eupatorium adenophorum and sericulture waste were used individually for the cultivation of Pleurotus sajor-caju, alone and mixed with paddy straw. P. sajor-caju, when used alone, exhibited a very good colonizing ability on these substrates, except in sericulture waste. The biological efficiency of P. deltoides and E. adenophorum when used as pure substrate was 75 and 77%, respectively, but it increased to 102% when P. sajor-caju was cultivated in a mixture with paddy straw in a ratio of 1:2. Experiments examining the growth on sericulture waste in both pure and mixed substrate are encouraging. From the analysis of substrate before and after the cultivation of P. sajor-caju it was noted that subsstrates were enriched in their protein content as a result of growth of this mushroom. The percentage of degradation of cellulose, hemicellulose and lignin showed that P. sajor-caju is capable of utilizing all three major components. The fruit bodies of P. sajor-caju were analyzed for crude protein content, crude fat and carbohydrate content. The energy values in the fruit bodies of P. sajor-caju and different organic wastes were found to vary from 282 to 309 kcal/100 g and from 319 to 467 kcal/100 g, respectively. It was found, however, that the energy recovery from organic wastes by fruit bodies was very low, i.e. 4.19-8.73 kcal/100g of dry substrate.

Notes: The potential for the production of 1,4-piperazinium-(L, L)-dilactate from L(+)-lactic acid preparations obtained by fermentation was studied. Piperazinium dilactate was found to be a very suitable source material for poly(lactic acid) production. In a novel polymerization process, the intermediate dilactide was directly formed in the salt melt at a moderate temperature. High-performance cultivation of Lactobacillus paracasei on a glucose-MRS medium was carried out using high-viability inocula. After the cell mass had been removed from the fermentation broth by centrifugation and/or ultrafiltration, the lactic acid solution was concentrated to 45% [w/w] by a two-stage electrodialysis process. Two methods of preparing 1,4-piperazinium dilactate were developed: the first from the medium-concentrated lactic acid (45%) and the second from a highly-concentrated lactic acid (85%) obtained by evaporation from the first one. Because there were no physical data on 1,4-piperazinium-(L, L)-dilactate in specialized literature, the pure product was characterized according to its solubility characteristics, melting point and spectroscopic analysis.

Notes: A total of 65 yeast strains were screened for their ability to grow and ferment lactose in a standard DURHAM tube test at 30 °C. Based on the kinetic parameters for lactose and whey lactose fermentations in shake flask cultures, the strain Candida psedotropicalis 65 was chosen for further studies.Some of the cultural parameters affecting ethanolic fermentations on lactose were standardized. At an initial lactose concentration of 100-120 g/l in the medium containing concentrated whey or lactose, at 40 °C and within 48 h, the selected strain reached an ethanol concentration of 41-59 g/l, an ethanol productivity of 1.3-3.0 g/l/h, a lactose consumption of 99%, an ethanol yield 0.4-0.49 g/g and a biomass yield of 0.027 g/g.

Notes: High temperature extrusion cooking of rye was used as a pretreatment for ethanol fermentation, and yeasts and bacteria were compared for their fermentation rates. Extrusion cooking caused, on average, a 7.5% increase in ethanol yield in comparison to autoclaved samples. The best results were achieved for grain with a moisture of 21-23% which was extruded at temperatures of 160-180 °C.Extrusion decreased the relative viscosity of rye grain water extracts, so it was possible to mash it without α-amylase. The efficiency of fermentation of extruded rye without Termamyl was equal to that of autoclaved and traditionally mashed rye (using α-amylase).The rate of fermentation of extruded rye grain by Zymomonas was higher during the first stage, but the final ethanol yield was similar for the bacterium and the yeast.Though both microorganisms gave good quality distillates, the concentration of compounds other than ethanol achieved from extruded rye mashes, which were fermented by Z. mobilis, was five times lower than for yeasts.

Notes: An altered ploidy level was observed in plants regenerated by adventitious shoot formation from seedlings of Hypericum prformatum L. (2n = 4x = 32). Among the somaclones of the Ro generation, the presence of diploids (2n = 2x = 16), triraploids (2n = 3x = 24), tetraploids (2n = 4x = 32) and mixoploids was detected. Cytogenetic analyses of the R1 and R2 progenies showed that the chromosomal instability of the Ro somaclones was transferred onto the next generation.While almost all the seed progeny of diploids (100% in R1 and 94% in R2) progenies showed that the chromosomal instability of the Ro somaclones was transferred onto the next generations.While almost all the seed progeny of diploids (100% in R1 and 94% in R2) and more than 60% of tetraploids (61% in R1 and 73% in R2) retained their chromosome number, cytogenetic diversity was observed in the progeny of triploids, mixoploids and some tetraploids.Somaclones and their offspring were analyzed for hypericin content. Statistical evaluation showed a correlation between hypericin content and ploidy during a two-year cultivation of R0 somaclones and in their R1 and R2 progenies.

Notes: Urban waste waters were treated with pure ozone or combinations of ozone, hydrogen peroxide and/or UV radiation to study the course of resulting BOD (biological oxygen demand)-time profiles and to propose a kinetic model. BOD-time profiles of chemically treated waste waters show an initial lag period that first order kinetic models cannot describe. A second order kinetic model is then proposed that satisfactorily fits experimental BOD-time profiles, except when hydrogen peroxide has been used. In these cases, BOD-time profiles present the highest lag periods observed. By applying this model, three parameters are determined: the biokinetic constant (k) which is an index of the biological removal rate; the potential amount of biodegradable matter (BODT), and the measure of the size of inocula and microbial activities of microoganisms (λ). The model was checked with experimental results of BOD-time profiles corresponding to both untreated and chemically ozonated urban waste waters. Ozonated waste waters showed the highest values of k and BODT, which implies an improvement of waste water biodegradability after ozonation. However, values of λ corresponding to ozonated waste waters presented lower values than those of untreated waste waters. This was due to the lag period observed in the BOD-time profile, which was a consequence of a lack of microorganism acclimation to ozonated waste waters. The effect of the ozone does, pH and carbonates during ozonation on COD (chemical oxygen demand) and the above indicated parameters was also studied. There was an optimum ozone dose which was 138 mg/l for this specific system. This led to the highest biodegradable fraction (ϕ) and the highest biokinetic constant (39% increase in ϕ and 4.7- fold increase in the value of k, respectively, compared to untreated waste waters.). Another significant fact was that a higher COD reduction was observed in the absence of carbonate during ozonation at basic pH values. In addition, the percentage of variation in the biodegradable fraction (Δϕ) of ozonated waste water increased compared to the untreated waste water at acid pH. The results suggest that ozonolysis, the direct molecular ozone way of reaction, due to its selective character, increases the biodegradability of waste water more than other chemically advanced oxidation processes based on hydroxyl radical reactions.

Notes: Waste waters from olive oil processing may cause severe pollution in the Mediterranean area, since they have a high level of chemical oxygen demand (COD) (100-200 g/l) and contain other organic and inorganic compounds. In all olive oil producing countries, the reduction of pollution in olive oil mill waste waters at reasonable costs and using techniques suitable for most industrial applications is an unsolved problem.For this paper, the yeast Yarrowia lipolytica ATCC 20255 was grown on waste waters from an olive oil mill in a 3.5 1 fermenter under batch culture conditions.The results showed that the yeast was capable of reducing the COD value by 80% in 24 h. In this way, a useful biomass of 22.45 g/l as single cell protein (SCP) and enzyme lipase were produced.During this process, most of the organic and inorganic substances were consumed, only aromatic pollutants were still present in the fermentation effluents. Therefore, we used a phenol degrader, namely Pseudomonas putida, to reduce phenolic compounds in the fermentation effuents after removing Yarrowia lipolytica cells. P. putida was effective in reducing phenols in only 12 h.

Notes: Methylobacterium rhodesianum MB 126 was cultivated using extended cultures without outflow. The feeding regime was based on the pH-regulated synchronous dosages of ammonia, methanol, phosphatc and trace elements according to supposed stoichiometric relations. The acidity of the culture medium was kept constant at pH 6.8, whereas the dissolved oxygen concentration was adjusted at 80% of saturation by autoregulation of the stirrer speed. However, besides testing technical conditions, two types of fermentations were discovered which are described in this paper. Firstly, although at the beginning of the bioprocesses the impeller speed increased up to 2,000 rpm, a decrease of dissolved oxygen down to zero was unavoidable. Secondly, methanol was accumulated temporarily up to 44 g/l and 26 g/l at 23 h of fermentation time and without inhibition of growth at least up to 30 g/l or PHB production. During this accumulation of the carbon substrate, exponential growth phases were detected showing growth rates of μ = 0.20/h and 0.21/h. But then, phases of retarded growth followed, whereas the methanol disappeared either continuously or after a steady level. In the course of a 54-h fermentation period, the synthesized PHB amounted to a content of above 50% of cell dry mass. From this data, a volumetric productivity of 0.4 g PHB/lxh was estimated. Moreover, the growth related yield coefficients were calculated to YX/MeOH = 0.21 and YX/MeOH = 0.14, whereas the product related yield coefficients amounted to YPHB/MeOH = 0.12 and YPHB/MeOH = 0,09. Since the shift down of growth rates as well as the production of PHB agreed in time with partial oxygen limitation (40% oxygen saturation), the competition observed between the tricarboxylic acid cycle and PHB synthesis was discussed. Summarizing the results, it was concluded that the frequently described inhibitory effect of methanol of above 2 g/l seems to be rather an effect of experimentally chosen conditions than of a general physiological phenomenon. Therefore, it could be demonstrated that the toxicity of methanol could be overcome if it was not dosed at different times but simultaneously with other medium components.

Notes: The influence of the precultivation with different carbon sources on the ability of three different bacterial strains (Sphingomonas sp. strain BA2, Gordona sp. strain BP9, Mycobacterium sp. strain VF1) to grow on phenanthrene. anthracene, pyrene or fluoranthene as the sole source of carbon and energy were studied. The strains were found to maintain their ability to grow on two of the four PAH after 30 serial transfers in liquid nutrient broth medium without selective pressure. The ability to grow on these PAH as the sole carbon and energy source was also maintained after curing experiments with acridine orange. The high stability of the PAH-degradation phenotype suggests that the tested strains carry at least parts of the PAH-degradation pathway genes on the chromosome. The PAH-degradation versatility of the strains was also influenced by the carbon source being used for precultivation. Possible reasons for the particularly good impact of the precultivation on hexadecane on the PAH degradation are discussed in this paper.

Notes: Studies of the properties of subtilisin, Novo type, immobilized on porous glass with the aid of hexamethylene diisocyanate were carried out. The immobilized proteinase preparation shows optimum activity at a pH value of 10.7 and at a temperature between 60-65°C. It was stable in a wider range of pH and temperature values than the native subtilisin. The KM values for hemoglobin and BAEE were 9.2 × 10-5 [M] and 139 × 10-5 [M], respectively. Under relatively non-aqueous conditions, immobilised subtilisin was able to synthesize phenylacetic acid ethyl ester.

Notes: A number of agricultural residues such as Saccharum munja Roxb. (Sarkanda), Oryza sativa L. (Paddy straw, as the control), Vinna unguiculata (L.) Walp (Cowpeas), Abelmoschus esculentum (L.) Moench (Lady's finger), Zea mays L. (Maize) and Cyamopsis tetragonoloba (L.) Taub. (Guar) were used for the cultivation of Pleurotus sajor-caju (Fr.) Singer. The biological efficiency of the fruit bodies of Pleurotus sajor-caju from the above mentioned substrates were found to be 13.47, 11.20, 8.37, 8.31, 6.87, and 6.08 percent, respectively. S. munja and paddy straw were found to be the best substrates for the growth of P. sajor-caju followed by Z. mays, V. unguiculata, A. esculentum, and C. tetragonoloba.

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Notes: The non-pollutant plant support material of the dwarf duckweed Wolffia arrhiza (Fam. Lemnaceae) was used for the entrapment of living yeast cells (Kluyveromyces fragilis) which hydrolyse lactose with the subsequent fermentation of glucose and galactose at high cell densities (up to 7.0 × 108/ml support). The stabile yeast-plant cell immobilizates are able to produce ethanol from lactose-containing media (e.g. whey) by batch fermentation (on a rotary shaker) or continuous fermentation (in a turbulence reactor) for several days (at a pH below 4.2 and a temperature of 30°C). The removal of whey proteins by a preceding heat denaturation of whey, high dilution rates, CSo values of 50 to 60 g lactose per litre whey and the preferential use of the K. fragilis strain DSM 7238 were determined as the prerequisites for an optimum continuous fermentation. Economically interesting productivities (Pmax ≥ 15 g ethanol/1 · h, D = 0.72 h-1) with an actual lactose turnover of 90% were obtained by using these parameters.

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Notes: Cell structured support material (CSM) prepared from Wolffia arrhiza fronds were loaded with Saccharomyces cereevisiae by the propagation of yeast cells introduced in the aerenchyma. The loading process could be strongly accelerated by preventing the growth of freely suspended yeasts in the fermenter. The pathway of the inoculation through the stomata could be visualized by transparent light microscopy of amylase and protease treated CSM. Slices of the loaded CSM clearly showed that the framework of the inner chlorenchyma cells remains intact during the propagation of the included yeast cells. Most of the yeast cells were found to be localized as a dense suspension within unbroken cells. During long-term treatment of the loaded CSM at high convection, about one third of the yeasts was released into water with a constant rate greater than 0.34 h-1. The residual amount of yeast cells remained in a stable compartment from which further loss did not occur for the period tested (20-48 h). It is assumed that the stable compartment is identical with the interior of chlorenchyma cells. The results are discussed in relationship to the possible use of the immobilized biomass in fermentation.

Notes: Growing Penicillium raistrickii i 477 cells immobilized by microencapsulation, entrapment in calcium alginate beads and photopolymerization were used for the 15α-hydroxylation of 13-ethyl-gon-4-en-3,17-dione (I) to 15α-hydroxy-13-ethyl-gon-4-en-3,17-dione (II). The immobilized cells had lower maximum specific growth rates and yield coefficients when cultivated on the carbon source glucose than the non-immobilized cells, which leads to lower volumetric productivities than the use of nonimmobilized cells. However, the cells immobilized by microencapsulation and calcium alginate entrapment showed a specific productivity equal to that of the respective non-immobilized cells based on product formation per dry biomass and time. Photommobilized cells were not able to grow in the presence of the steroid because the substrate concentrations within the polymer reached inhibiting amounts for growth and product formation. In the absence of the steroid, the growing photoimmobilized cells showed a prolonged lag-phase in comparison with the free cells.

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Notes: Bioprocess engineering at present concentrates on the enormous problems in the environment. What is therefore needed is a sound methodology, which should be based on the interactions between the physiology of biological reaction networks and physical processes in the environment and which should be an analogy to bioreactor performance. The conventional methodology is empirically oriented, using pilot plant data for the experimental estimates of process economics, where all further details are elucidated following the mechanistic approach on the microscopic level based on assumed mechanisms (causalities). According to the new view, the new systems-based methodology uses mathematical models as approximations and includes all the interactions. Pilot plant data are needed for model falsification, using analogies on the formal macroscopic level. Bioreactor scale-up as one application is a more rapid procedure of reasonable accuracy, where both the biokinetics as well as the fluid dynamics are quantified using formal macroscopic analogies.Model consistency and plausibility are the basic criteria when using model computer simulations as a decisive aid, while experiments lose their central role and are on longer the basis for evaluating the scientific work; they are simply the basis of the researcher's intuition. Another typical feature of complex systems is that model parameters are interdependent. The final decisive fact will be the mental experiment (“thinking” with the left and right side of the brain), which can be supported using computer simulations. This evolutionary interplay between the three realities of thinking, experimenting and simulating leads to a holistic progress towards the better understanding of highly complex systems. It offers the solution to the problems, which is needed in future.

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Notes: Penicillium notatum No. 1 as a producer of β-galactosidase was cultivated in a 5-1 fermenter. Various methods of protein isolation and concentration from the culture fluid were optimized. Then the conditions of β-galactosidase purification using an affinity chromatographic technique were established. The purified enzyme was immobilized on a controlled porous glass (CPG). The optimum temperature and pH values of the native and immobilized forms of β-galactosidase were determined as 50°C and 30-50°C as well as pH 3 and pH 3-5, respectively.

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Notes: A review of the intensification of moment, mass and heat transfer using an “unconventional” method, namely the application of static mixers, also known as “motionless mixers”, is presented in this paper.After the presentation of some structural types of static mixers, studies with particular emphasis on flow behaviour, pressure drop, mixing, mass and, heat transfer in the presence of static mixing devices, used in biotechnology and chemical engineering, were discussed.The suitability of static mixers to enhance the transfer process parameters was also justified by the low energy requirement, while the fluids might have different viscosities and because of this fact they could remove any mistakes made by the equipment.The article is not directed towards a comprehensive review, but it should serve as a landmark in future undertakings.

Notes: The development of model plant-microbial associations between Gram negative soil microbes capable of degrading phenoxyacetate herbicides, such as 2,4-D and 2,4-D methyl ester, and the crops canola and wheat was described. Both an Acinetobacter baumannii pJP4 transconjugant and Alcaligenes eutrophus JMP 134 colonised non-parasitically on the roots of sterilised seedlings in a hydroponic system. Laser scanning confocal microscopy has shown that colonisation occurred both on the root surface and deeper inside the mucilage layer or inside some surface root cells. When 2,4-D was added to the hydroponic medium supporting the growth of those seedlings colonised by 2,4-D degrading bacteria, the gas chromatographic analysis showed a rapid decrease in the concentration of this herbicide. These bacteria colonising the root system were shown to be responsible for the degradation of 2,4-D. Plants inoculated with the 2,4-D degrading microbes were subsequently found to be less susceptible to damage by the herbicide in such hydroponic systems.

Notes: Under the conditions of various aeration and medium mixing intensities (even at a constant medium oxygen partial pressure value), the variations in the RNA synthesis as well as the growth rate of lysine producing Brevibacterium flavum strains are inversely correlated with the ppGpp concentration in the cells.An increase in the ppGpp synthesis and a decrease in the RNA content in the cells was observed in the cases of a low cell energy charge value (lower than 0.6). This took place in the cases of bacterial cultivation at a low or very high medium aeration and mixing intensity. Hence, the energy production in the cells, the ppGpp synthesis and the growth control mechanism in Corynebacteria may be regarded as connected processes.

Notes: Minute nuclei named “smaller nuclei” were generated when the cells of Saccharomyces cerevisiae were treated with colchicine. The formation of “smaller nuclei” seemed to be related to nuclear division because those nuclei were only produced under conditions suitable for nuclear division. The fact that the average DNA content of “smaller nuclei” was almost one tenth of that of the isolated normal diploid nuclei showed that the “smaller nuclei” are not condensed nuclei but aneuploid nuclei like micronuclei in animal cells. It appeared therefore likely that a micronuclei-like structure could be produced by colchicine treatment in S. cerevisiae.

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Notes: Lignin was mineralized in the experiments in which 14C-lignin was incubated with lignin peroxidase or manganese peroxidase in a tartrate buffer in the presence of cycloheximide-treated protoplasts obtained from the ligninolytic mycelia of Phanerochaete chrysosporium. The rate of lignin mineralization was dependent on the lignin peroxidase or manganese peroxidase concentration in the medium. In the experiments in which lignin was incubated with lignin peroxidase or manganese peroxidase, lignin was repolymerized irrespective of the presence of protoplasts mineralizing lignin, suggesting that an active degradation of lignin and repolymerization took place. Taking into account that lignin peroxidase and manganese peroxidase were the only extracellular enzymes in the experiments in which lignin was mineralized by the protoplasts, it is postulated that lignin peroxidase and/or manganese peroxidase can degrade lignin into small fragments which can then be further absorbed by the fungal cells and subsequently degraded to CO2.

Notes: The behaviour of different Lactobacillus casei and Lactobacillus plantarum species in the fermentation of Manchego whey was experimentally studied and the results were statistically analyzed using a hypothesis contrast method. The steadiness of the velocity of the production of lactic acid during the fermentation process allowed the use of this variable to compare the different microorganisms. From this comparison it was inferred that the individuals of the same population behave alike and that the L. casei population produces lactic acid at a higher rate than the L. plantarum population. A competitive effect among the members of the L. casei population was also observed.

Notes: The present paper reviews the different monoterpenols and monoterpene polyols identified in the glycoside form in the plant kingdom. The glycosidic moieties involved are also considered. The natural pathways for the synthesis and hydrolysis of these monoterpene glycosides and the different hypothesis concerning their metabolism are discussed. The present state of different projects for the biotechnological transformation of these aromatic precursor compounds is reported.

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Notes: The production of extracellular glucose oxidase in a submerged culture by a number of auxotrophic, 2-deoxy-D-glucose resistant and protease-less mutants of Aspergillus niger was evaluated. Among the auxotrophic strains, no evident dependence was found between the kind of the nutritional requirements and the level of the glucose oxidase activity. However, the majority of auxotrophs, requiring serine or niacin, showed a higher enzyme activity (from 16 to 680%) than the parent strain. The dynamics of the glucose oxidase synthesis by the free and immobilized mycelium of the most active niacin- mutant of A. niger was also investigated.

Notes: The 5C outer membrane protein, one of the N. meningitidis class 5 proteins, was preferably expressed in bacteria isolated from the nasopharynx and its role in adhering to the mucosal cells and invading them as well as the development of anti-5C antibodies in healthy carriers was demonstrated. Anti-5C monoclonal antibodies are bactericidal in the presence of the human complement. The immunodominant region of the 5C protein is highly conserved among the different strains of N. meningitidis, and the opc gene, which encodes the protein, does not seem to show antigenic variations.Here the isolation of the opc gene from the Cuban strain B:4:P1.15 by PCR (Polymerase Chain Reaction) is presented. Under the regulation of the tryptophan promoter, the gene was cloned and sequenced in E. coli with a high level of expression and fused to the amino-terminal end of the interleukin-2 gene. In the dot-blot experiments, the presence of the gene in those strains which did not express the protein in the whole cell ELISA was also detectable.

Notes: A simple medium enhancing the production of thiaminase I (EC 2.5.1.2) by Bacillus thiaminolyticus was developed. Ca2+ stimulated the enzyme production. The activity of extracellular thiaminase I ranged between 1.29 and 1.33 U/ml medium.

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Notes: The production of cellulase was investigated in semi-solid state culture using the immobilized mycelium of Trichoderma reesei mutants on polyurethane foam impregnated with lactose medium. An extremely high value of about 2.6 FPU/ml was reached after the cultivation of T. reesei D-78085 on a 0.5% lactose medium in continuous culture at a pH medium of 4.0 when a bioreactor with vertical polyurethane foam plates was used. The enzyme yield on lactose was 520 FPU/g of lactose metabolized in comparison with 160 FPU/g using a stirred tank bioreactor.

Notes: The nitrilase of Rhodococcus rhodochrous PA-34 catalyzes the production of optically active amino acids from aminonitriles. The amino acid sequence of the NH2 terminus of the purified nitrilase was determined for the preparation of a synthetic oligonucleotide as a southern hybridization probe. A 9.5-kbp Pst I-fragement, which hybridized with the oligonucleotide probe, was isolated from R. rhodochrous PA-34 genomic libraries constructed in pUC 19. Nucleotide sequence analysis revealed that the nitrilase gene codes for a putative polypeptide of 380 amino acids which correspond to a relative molecular weight of 41, 723.

Notes: Cephalosporin C biosynthesis was investigated in a pilot-plant external-loop airlift bioreactor for evaluating the capacity of this bioreactor to surpass the problems which arise from the morphology of the mould and the rheology of the broth.Some of the results were compared with those obtained in a stirred tank bioreactor.The dilution and the use of static mixers was necessary to overcome the effects of the high viscosities. The oxygen transfer rate represented 84% of that in the stirred bioreactor, but the efficiency of the power utilization was higher.The specific productivity of Cephalosporin C is comparable to that obtained in the stirred tank bioreactor, but the average specific power consumption was found to be 2/3 of that in the stirred vessel.

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Notes: The influence of two mixing systems on the principal parameters of mycelial fermentations of Aspergillus niger, Fusicoccum amygdali Del. and Fusarium moniliforme Sheld. as well as their metabolite citric acid, fusicoccin and gibberellic acid production was analyzed from the viewpoint of flow energy distribution in a bioreactor.The growth and metabolite synthesis during fermentation was compared under different mixing conditions in the fermenter FU-8 with a turbine mixing system (TMS) and a counterflow mixing system (CMS). It was found that the growth, productivity and respiration characteristics as well as the morphology of these cultures varied dependent on the mixing system and agitation regime used. The counterflow mixing system was more favourable for large agglomerates (F. amygdali) or soft pellets (A. niger) forming fungi, while the turbine mixing system was more effective for F. moniliforme growing in the form of small clumps and freely dispersed hyphae. Flow characteristics under different mixing conditions were analyzed in a model fermenter.The kinetic energy of flow fluctuations was measured in gassed and ungassed water and different fermentation broth systems by using a Stirring Intensity Measuring Device (SIMD-F1). The difference of the energy values at different points was better expressed in the fermenter with a turbine mixing system in comparison with that having a counterflow mixing system. High viscous F. amygdali and A. niger broth provided higher energy values compared to water and low viscous F. moniliforme broth. It was observed that the intensity of growth and the intensity of the synthesis decreased at very high energy values, which was obviously connected to the influence of the irreversible shear stress on the mycelial morphology.

Notes: Permeabilized cells of Kluyveromyces marxianus CCY eSY2 were tested as the source of lactase in the ethanol fermentation of concentrated deproteinized whey (65-70 g/l lactose) by Saccharomyces cerevisiae CCY 10-13-14. Rapid lactose hydrolysis by small amounts of permeabilized cells following the fermentation of released glucose and galactose by S. cerevisiae resulted in a twofold enhancement of the overall volumetric productivity (1.03 g/l × h), compared to the fermentation in which the lactose was directly fermented by K. marxianus.

Notes: The stability behaviour of the pBR322 plasmid derivative pBB210 with β-lactamase gene and human interferon-α1 gene in Escherichia coli TG1 was studied in chemostat cultures under non-selective (medium without antibiotics), selective (medium with β-lactam antibiotic ampicillin) and modified selective (medium with ampicillin and the β-lactamase inhibitor sulbactam) conditions. Under non-selective conditions, a behaviour typical of unstable systems was found. Under selective conditions, the behaviour predicted by the models was obtained - the fraction of plasmid-bearing cells in the population approached a constant value which was dependent on the ampicillin concentration in the feeding and on the cell concentration in the chemostat. Under modified selective conditions, the higher the concentration of sulbactam in the medium was, the higher the fraction of plasmid-bearing cells was in steady state conditions.

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Notes: The effect of the rapid reduction of the water activity (aw) on the extracellular protein and amylolytic activity of Aspergillus niger was studied. An aw value gradient from 0.90 to 0.99 in KCl solutions was applied for the mycelium treatment. It was found that the aw reduction considerably influenced the protein secretion. This phenomenon was dependent on the age of the treated mycelium and the range of the aw gradient. The highest protein and enzyme secretion yields were obtained at aw = 0.98 using a 72-h old mycelium. In comparison with the non-treated mycelium, the increase in the secretion amounted to about 60% for the amylolytic activity and 37% for the soluble protein, respectively. It was shown that the mycelium incubated in KCl solutions of an aw value from 0.90 to 0.99 had the ability for regeneration in fresh CZAPEK-DOX medium. The effect of the osmotic shock on the protein secretion was limited only for the treated cell population and declined in the mycelium which was regenerated after the transfer into the culture medium.

Notes: Surface-active compound of biological origin (biosurfactants) have only been described in the past few decades. With the advantage of biodegradability and production on renewable resources, biosurfactants have been gaining prominence and their applications are becoming wider. So far, literature contains mixed reports on the successes of the applications of biosurfactants and their economical viability. They remain compounds which are not very well understood, yet, with several important applications. The target industries for biosurfactant use are the petroleum remediation industries and environmental conservation agencies. These industries, however, seem reluctant to use them for fear of dealing with microbes or microbial products. This includes cleaning up oil spills from the environment, remediation of metal-contaminated soils or waste streams, mobilizing heavy oil sludge and enhanced oil recovery. The importance of biosurfactants, their production, characteristics and limited successes and applications in oil pollution remediation and oil storage tank cleaning are discussed.

Notes: Leachate from a municipal waste landfill site was treated using an activated sludge bioreactor, a fluidized bed biofilm reactor and a packed-bed column reactor (trickling filter).The leachate contained high organic matter (2.0-2.6 g/l of COD), high ammonium (300-700 mg/l) and sulphide (200-800 mg/l) concentrations, as well as low metal concentrations.The continuously operating reactors were employed to study the effects of TOC loading on the removal of TOC as well as on the nitrification and denitrification processes.Among the three biological treatment technologies investigated, the fluidized bed biofilm reactor was best with respect to removing ammonia and TOC. More than 90% of TOC and 99% of ammonia were removed when TOC loading was less than 0.5 kg/m3 × d. At a TOC loading of 4 kg/m3 × d, the removal of TOC and ammonia was 80% and 99%, respectively.In contrast, the treatment of leachate with the packed-bed reactor was successful in TOC removing only at TOC loading less than 0.3 kg/m3 × d (TOC elimination decreased from 86% at 0.06 kg/m3 × d to 60% at 0.3 kg/m3 × d). However, the reactor was active in nitrification even at a higher TOC loading (more than a 98% ammonia elimination at a TOC loading of 0.5 kg/m3 × d).Leachate was processed in the activated sludge reactor when TOC loading was less than 0.5 kg/m3 × d (with a removal of TOC and ammonia up to 83% and 99%, respectively). The activated sludge reactor was also effective in TOC removal at a higher TOC loading (e.g. a 74% TOC removal at a TOC loading of 1 kg/m3 × d), but for ammonia elimination, the activity continuously decreased (less than 60% ammonia removal at a TOC loading of 1 kg/m3 × d).Overloading in the activated sludge system was indicated by a high concentration of ammonia and nitrite in the effluent. In the packed bed reactor, overloading was characterized by a progressively incomplete TOC removal. No significant overloading was found in the fluidized bed reactor up to a TOC loading of 4 kg/m3 × d.

Notes: No Abstracts.