Microbubble intensification of bioprocessing.

Microbubbles have been involved in industrial processing since the 1970s with the introduction of dissolved air flotation into common practice. The turn of the century saw microbubbles become regularly used in medical imaging. But in bioprocessing, only this decade has seen rapid advances in R&D, with some bioprocesses, particularly in wastewater treatment, adopted at full industrial scale, and others at pilot scale, such as anaerobic digestion and fermentation, which is full industrial scale for many biomanufacturing and pharmaceutical processes. This article reviews the methods of microbubble generation only briefly, as it turns out only one generation method, fluidic oscillation through microporous diffusers, has the requisite features for introduction into full scale fermentation processes. Subsequently, six fundamental physicochemical hydrodynamics mechanisms that have been exploited by microbubble innovations in bioprocessing are presented and analyzed, particularly for the roles they play in bioprocessing applications. Some examples are drawn with applications to microalgal and yeast processing, as well as usage in wastewater treatment processes. Because the smallest microbubbles can increase rates in some of these six fundamental processes by several orders of magnitude over conventional processing methods, with the optimal contacting patterns, the promise for wider exploitation in bioprocessing is substantial.

Factors influencing exfiltration processes in sewers.

Exfiltration from sewers is widespread and emerging legislation may require water service providers to identify, and rectify, its sources in sewerage systems. This paper describes exfiltration test apparatus and a series of experiments undertaken using sewage to gain a better understanding of the influence of sewage solids and sediments on leakage rates. An overview of the results obtained is given, which demonstrates that most previous estimates of exfiltration leakage rates were too high due to a lack of appreciation of the "self-repairing" action of sewage and sewage associated solids. Exfiltration rates of 0.1% of the sewer flow or 0.001 I/s have been recorded for defects up to 6 mm wide.

Gene-specific expression and calcium activation of Arabidopsis thaliana phospholipase C isoforms.

• PI-PLCs synthesise the calcium releasing second messenger IP3 . We investigated the expression patterns of the Arabidopsis PI-PLC gene family and measured in vitro activity of encoded enzymes. • Gene specific RT-PCR and promoter-GUS fusions were used to analyse AtPLC gene expression patterns. The five available AtPLC cDNAs were expressed as fusion proteins in Escherichia coli. • All members of the AtPLC gene family were expressed in multiple organs of the plant. AtPLC1, and AtPLC5 expression was localized to the vascular cells of roots and leaves with AtPLC5::GUS also detected in the guard cells. AtPLC4::GUS was detected in pollen and cells of the stigma surface. In seedlings, AtPLC2 and AtPLC3 were constitutively expressed, while AtPLCs 1, 4 and 5 were induced by abiotic stresses. AtPLC1-5 were all shown to have phospholipase C activity in the presence of calcium ions. • AtPLCs showed limited tissue specific expression and expression of at least three genes was increased by abiotic stress. The differing calcium sensitivities of recombinant AtPLC protein activities may provide a mechanism for generating calcium signatures.

Big bacteria pass through very small holes.

Can normal-sized bacteria pass through small holes, and if so how? We have found that common, potentially pathogenic, bacteria (which are nominally larger than 0.2 microm) can cross a 0.2 microm nylon membrane. All of the bacteria crossed from the upper membrane surface to the solid medium below the membrane; this ability was highly repeatable and did not depend on the make of membrane used. Bacteria growing below the membrane exhibited normal size and morphology. We suggest that the ability of common bacteria to pass through small holes has important implications in relation to their role as pathogens.

Crystallization and preliminary X-ray analysis of glucose dehydrogenase from Haloferax mediterranei.

Glucose dehydrogenase (E.C. 1.1.1.47; GlcDH) from Haloferax mediterranei has been overexpressed in Escherichia coli, solubilized by the addition of 8 M urea and refolded by rapid dilution. The protein has been purified by conventional techniques and crystallized by the hanging-drop vapour-diffusion method using sodium citrate as the precipitant. Two crystal forms representing the free enzyme and the binary complex with NADP(+) grow under these conditions. Crystals of form I diffract to beyond 3.5 A resolution and belong to the hexagonal space group P622, with unit-cell parameters a = b = 89.1, c = 214.6 A, alpha = beta = 90, gamma = 120 degrees. Crystals of form II diffract to greater than 2.0 A and belong to the orthorhombic space group I222 or I2(1)2(1)2(1), with unit-cell parameters a = 61.8, b = 110.9, c = 151.7 A, alpha = beta = gamma = 90 degrees. Calculated values for V(M) and consideration of the packing for both crystal forms suggests that the asymmetric units in both crystal forms contain a monomer.

Mechanisms for controlling balance between light input and utilisation in the salt tolerant alga Dunaliella C9AA.

The yield of photosynthetic O2 evolution was measured in cultures of Dunaliella C9AA over a range of light intensities, and a range of low temperatures at constant light intensity. Changes in the rate of charge separation at Photosystem I (PS I) and Photosystem II (PS II) were estimated by the parameters ΦPS I and ΦPS II . ΦPS I is calculated on the basis of the proportion of centres in the correct redox state for charge separation to occur, as measured spectrophotometrically. ΦPS II is calculated using chlorophyll fluorescence to estimate the proportion of centres in the correct redox state, and also to estimate limitations in excitation delivery to reaction centres. With both increasing light intensity and decreasing temperature it was found that O2 evolution decreased more than predicted by either ΦPS I or ΦPS II. The results are interpreted as evidence of non-assimilatory electron flow; either linear whole chain, or cyclic around each photosystem.

The effect of ionic stress on photosynthesis in Dunaliella tertiolecta : Chlorophyll fluorescence kinetics and spectral characteristics.

A comparison of the effects of ionic stress and an uncoupler on long-term fluorescence transients (the 'Kautsky effect') in the green alga Dunaliella tertiolecta indicated that the large quenching induced by ionic stress was caused by a pH gradient across the thylakoid membrane. This possiblity was given support by the increase in the slow phase of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in algae subjected to ionic stress. Low-temperature fluorescence emission spectra indicated that salt stress enhanced photosystem-I emission in the dark, and a comparison of simultaneous emissions at 695 and 720 nm at room temperature indicated a further increase in photosystem-I emission during the fluorescence transients. Taken together with the decrease in the fast phase of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in stressed algae, our results indicate that ionic stress stimulates cyclic electron flow, and that non-cyclic flow is inhibited. The effect of sucrose-induced osmotic stress was similar to, but less marked than, the effects of NaCl and KCl; the effect of decreasing the external salinity was small.