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The high-energy input for harvesting biomass makes current commercial microalgal biodiesel production economically unfeasible. A novel harvesting method is presented as a cost and energy efficient alternative: the bio-flocculation by using one flocculating microalga to concentrate the non-flocculating microalga of interest. Three flocculating microalgae, tested for harvesting of microalgae from different habitats, improved the sedimentation rate of the accompanying microalga and increased the recovery of biomass. The advantages of this method are that no addition of chemical flocculants is required and that similar cultivation conditions can be used for the flocculating microalgae as for the microalgae of interest that accumulate lipids. This method is as easy and effective as chemical flocculation which is applied at industrial scale, however in contrast it is sustainable and cost-effective as no costs are involved for pre-treatment of the biomass for oil extraction and for pre-treatment of the medium before it can be re-used.

The toxicity of homologous series of organic solvents has been investigated for the gram-positive bacteria, Arthrobacter sp. and Nocardia sp., and the gram-negative bacteria, Acinetobacter sp. and Pseudomonas sp. The hydrophobicity of the solvent, expressed by its logP(octanol), proves to be a good measure for the toxicity of solvents in a two-phase system. The transition from toxic to nontoxic solvents occurs between logP(octanol) 3 and 5 and depends on the homologous series. No correlation has been found between the hydrophobicity of the substituent on the alkyl backbone of the solvent and the location of the transition point in toxicity. The logP(octanol), above which all solvents are nontoxic, is used to express the solvent tolerance of the bacteria. In general, the solvent tolerance of gram-negative bacteria is found to be slightly higher than that of gram-positive bacteria, but this does not hold for all homologous series of organic solvents investigated.Because the toxicity effects of organic solvents in a two-phase system can be ascribed to molecular as well as phase toxicity effects, molecular toxicity effects were investigated separately in a one-phase system with subsaturating amounts of organic solvent. The solvent concentration in the aqueous phase, at which 50% of the metabolic activity of the bacteria is lost, is used to express solvent toxicity. This concentration is found to be similar for the gram-positive Arthrobacter and the gram-negative Acinetobacter. Assuming the critical membrane concentration theory (G. J. Osborne et al. Enzyme Microb. Technol. 1990, 12: 281-291) to be valid, it can be concluded that differences in solvent tolerance between these two bacteria, cannot be ascribed to differences in response to molecular toxicity. Prediction of the toxicity of any solvent, using the critical membrane theory, appears to be possible in the case of alkanols or alkyl acetates. However, prediction of the toxicity of ethers appears to be impossible.

Simultaneous production and selective extraction of beta-carotene from living cells of Dunaliella salina in a two-phase system of aqueous and organic phases has been investigated. Solvents with values of log P(octanol), which denotes hydrophobicity of a compound, ranging from 3 to 9 were used as organic phase. Viability and activity of Dunaliella salina in the presence of organic solvents were checked by microscopic observation and photosynthetic oxygen-production-rate measurements, respectively. Extraction ability of different solvents for both beta-carotene and chlorophyll was determined spectrophotometrically. In addition, beta-carotene contents of the cells growing in the aqueous phase and extracted beta-carotene by the different organic phases were quantified by the same method. Results showed that solvents having log P(octanol) > 6 can be considered biocompatible for this alga. Moreover, pigment extraction ability of a solvent is inversely dependent on its log P(octanol) value. By increasing the degenerative hydrophobicity the extraction ability for both chlorophyll and beta-carotene, decreases. However, this decrease is more profound for chlorophyll. Therefore, selective extraction of beta-carotene becomes feasible. Comparison of the total beta-carotene produced in the presence and in the absence of solvents shows that the presence of a second phase of biocompatible solvents in the culture media may induce the beta-carotene production pathway. The beta-carotene productivity per cell in a two-phase system with dodecane was the highest observed. Extraction ability of the biocompatible solvents dodecane, tetradecan, and hexadecane was similar.

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In the literature measurements of kinetic data of enzymes in reversed micelles have been interpreted in two ways. In the first, all enzyme parameters are expressed with respect to the total volume of the reversed micellar solution. In the second, the enzymatic conversion is related only to the fraction of the volume consisting of aqueous solution (pseudophase model). In this paper equations are derived describing the rate of an enzymatic reaction for three different kinds of enzymes : enzymes obeying Michaelis-Menten kinetics, enzymes following a ping-pong bi-bi mechanism and enzymes which convert substrates according to an ordered mechanism. In deriving these equations, a distinction is made between intermicellar exchange reactions of substrate(s) and product(s) and the enzymatic reaction which takes place in the waterpool of a reversed micelle. In the description, all intrinsic rate constants of the enzyme are assumed to be independent of its environment. The rate equations show that the presence and efficiency of the intermicellar exchange reaction, which supplies the enzyme with substrate and removes product, can affect the rate of an enzymatic reaction under common experimental conditions.Whereas kinetic parameters derived from double-reciprocal plots often seem to be affected by enclosure in reversed micelles, these apparent deviations from kinetics in aqueous media can be explained by the model presented here as arising from exchange phenomena. Neither the experimentally determined maximum enzyme velocity, v , , , , nor the Michaelis constants are affected by the incorporation of the enzyme in reversed micelles.The deviations of kinetic parameters from the aqueous values are shown to depend strongly on the concentration of reversed micelles, the intermicellar exchange rate and the volume fraction of water, a dependence in agreement with findings reported in the literature.Reversed micelles are 1 -10-nm-diameter water droplets, dispersed by means of a surfactant, alone or in combination with a cosurfactant, in an organic solvent. The layer of surfaceactive compounds protects the enzymes in the water droplets from the adverse effects of the organic solvent. It has been well established that enzymes can be incorporated into reversed micelles in an active form (for reviews see [l, 21). Enzymes in reversed micelles can convert polar and apolar substrates. For polar substrates, a process of collision followed by opening up of the surfactant layer and mixing of the waterpools of substrate-and enzyme-filled reversed micelles enables substrate supply and product removal [3,4]. Effects of the enclos- ' s -I ) ; kin, rate of transport from a solute from the organic phasc into the reversed micelle; k,,,, rate of transport from a solutc from waterpool to the organic phasc; [MI, concentration of reversed micelles; m, occupation number of a substrate or a product in an enzyme and substrate-or product-filled reversed micelle; %', weighted average substrate or product occupation number in reversed micelles containing bot...

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