Continuous rhamnolipid production using denitrifying Pseudomonas aeruginosa cells in hollow‐fiber bioreactor

Rhamnolipids are high‐value effective biosurfactants produced by Pseudomonas aeruginosa. Large‐scale production of rhamnolipids is still challenging especially under free‐cell aerobic conditions in which the highly foaming nature of the culture broth reduces the productivity of the process. Immobilized systems relying on oxygen as electron acceptor have been previously investigated but oxygen transfer limitation presents difficulties for continuous rhamnolipid production. A coupled system using immobilized cells and nitrate instead of oxygen as electron acceptor taking advantage of the ability of P. aeruginosa to perform nitrate respiration was evaluated. This denitrification‐based immobilized approach based on a hollow‐fiber setup eliminated the transfer limitation problems and was found suitable for continuous rhamnolipid production in a period longer than 1,500 h. It completely eliminated the foaming difficulties related to aerobic systems with a comparable specific productivity of 0.017 g/(g dry cells)‐h and allowed easy recovery of rhamnolipids from the cell‐free medium. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29: 346–351, 2013     

Vegetation,herbivores and fires in savanna ecosystems: A network perspective

The dynamics of savanna ecosystems depends on the interplay between multiple factors such as grazing, browsing, fires, rainfall regime and interactions between grass and woody vegetation. In most modelling applications this interplay may not be fully understood because some of these drivers enter the models as dynamically independent factors. In this paper we consider such factors as dynamic variables. To analyze their interplay we focus on the structure of the interactive network of variables and exploit the properties of signed digraphs using the algorithm of Loop Analysis. Qualitative signed digraphs for the savanna ecosystem are developed and their predictions used to interpret patterns of abundance observed in case studies selected from the literature. The outcomes of this exercise unveil that: 1) the structure of the interactions is appropriate locus for the explanation of patterns observed in savannas; 2) signed digraph can help disentangling causative mechanisms by linking correlation patterns, source of change and network structure. This study highlights that central to the understanding of savanna dynamics is our ability to diagram the important relationships and understand how they interrelate with sources of variations to cause ecosystem change.     

Monitoring gradient formation in a jet aerated bioreactor

Jet aerated loop reactors (JLRs) provide high mass transfer coefficients (k_La) and can be used for the intensification of mass transfer limited reactions. The jet loop reactor achieves higher k_La values than a stirred tank reactor (STR). The improvement relies on significantly higher local power inputs (～10⁴) than those obtainable with the STR. Operation at high local turnover rates requires efficient macromixing, otherwise reactor inhomogeneities might occur. If sufficient homogenization is not achieved, the selectivity of the reaction and the respective yields are decreased. Therefore, the balance between mixing and mass transfer in jet loop reactors is a critical design aspect. Monitoring the dissolved oxygen levels during the turnover of a steady sodium sulfite feed implied the abundance of gradients in the JLR. Prolonged mixing times at identical power input and aeration rates (～100%) were identified for the JLR in comparison to the STR. The insertion of a draft tube to the JLR led to a more homogenous dissolved oxygen distribution, but unfortunately a reduction of mixing time was not achieved. In case of increased medium viscosities as they may arise in high cell density cultivations, no gradient formation was detected. However, differences in medium viscosity significantly altered the mass transfer and mixing performance of the JLR.     

Same but different–Scale up and numbering up in electrobiotechnology and photobiotechnology

Facing energy problems, there is a strong demand for new technologies dealing with the replacement of fossil fuels. The emerging fields of biotechnology, photobiotechnology and electrobiotechnology, offer solutions for the production of fuels, energy, or chemicals using renewable energy sources (light or electrical current e.g. produced by wind or solar power) or organic (waste) substrates. From an engineering point of view both technologies have analogies and some similar challenges, since both light and electron transfer are primarily surface‐dependent. In contrast to that, bioproduction processes are typically volume dependent. To allow large scale and industrially relevant applications of photobiotechnology and electrobiotechnology, this opinion first gives an overview over the current scales reached in these areas. We then try to point out the challenges and possible methods for the scale up or numbering up of the reactors used. It is shown that the field of photobiotechnology is by now much more advanced than electrobiotechnology and has achieved industrial applications in some cases. We argue that transferring knowledge from photobiotechnology to electrobiotechnology can speed up the development of the emerging field of electrobiotechnology. We believe that a combination of scale up and numbering up, as it has been shown for several photobiotechnological reactors, may well lead to industrially relevant scales in electrobiotechnological processes allowing an industrial application of the technology in near future.     

Use of image analysis to understand enzyme stability in an aerated stirred reactor

Efficient regeneration of NAD(P)⁺ cofactors is essential for large-scale application of alcohol dehydrogenases due to the high cost and chemical instability of these cofactors. NAD(P)⁺ can be regenerated effectively using NAD(P)H oxidases (NOXs) that require molecular oxygen as a cosubstrate. In large-scale biocatalytic processes, agitation and aeration are needed for sufficient oxygen transfer into the liquid phase, both of which have been shown to significantly increase the rate of enzyme deactivation. As such, the aim of this study was to identify the existence of a correlation between enzyme stability and gas–liquid interfacial area inside the bioreactor. This was done by measuring gas–liquid interfacial areas inside an aerated stirred reactor, using an in situ optical probe, and simultaneously measuring the kinetic stability of NOXs. Following enzyme incubation at various power inputs and gas-phase compositions, the residual activity was assessed and video samples were analyzed through an image processing algorithm. Enzyme deactivation was found to be proportional to an increase in interfacial area up to a certain limit, where power input appears to have a higher impact. Furthermore, the presence of oxygen increased enzyme deactivation rates at low interfacial areas. The areas were validated with defined glass beads and found to be in the range of those in large-scale bioreactors. Finally, a correlation between the enzyme half-life and specific interfacial area was obtained. Therefore, we conclude that the method developed in this contribution can help to predict the behavior of biocatalyst stability under industrially relevant conditions, concerning specific gas–liquid interfacial areas.     

Effect of non-feeding period length on the intermittent operation of UASB reactors treating dairy effluents

Recent environmental concerns have prompted a re-evaluation of conventional management strategies and refueled the search of innovative waste management practices. In this sense, the anaerobic digestion of both fat and the remaining complex organic matter present in dairy wastewaters is attractive, although the continuous operation of high rate anaerobic processes treating this type of wastewaters causes the failure of the process. This work accesses the influence of non-feeding period length on the intermittent operation of mesophilic UASB reactors treating dairy wastewater, in order to allow the biological degradation to catch up with adsorption phenomenon. During the experiments, two UASB reactors were subject to three organic loading rates, ranging from 6 to 12 g(COD) x L(-1) x d(-1), with the same daily load applied to both reactors, each one with a different non-feeding period. Both reactors showed good COD removal efficiencies (87-92%). A material balance for COD in the reactors during the feeding and non-feeding periods showed the importance of the feedless period, which allowed the biomass to degrade substrate that was accumulated during the feeding period. The reactor with the longest non-feeding period had a better performance, which resulted in a higher methane production and adsorption capacity for the same organic load applied with a consequent less accumulation of substrate into the biomass. In addition, both reactors had a stable operation for the organic load of 12 g(COD) x L(-1) x d(-1), which is higher than the maximum applicable load reported in literature for continuous systems (3-6 g(COD) x L(-1) x d(-1)).     

Microbial biofilms: new catalysts for maximizing productivity of long-term biotransformations

The performance of biocatalytic reactions is often hampered by product and/or substrate toxicity and short-term reaction times due to instable biocatalysts. Microbes in biofilms show a remarkable resistance against biocides and form stable communities. In nature, especially in environments characterized by harsh conditions such as heavily contaminated sites, cells grow pre-dominantly in biofilms, which enable them to cope with physiological stress. This robustness was utilized to design a bioprocess concept based on catalytic biofilms for stable long-term transformations of toxic reactants. Sixty-nine bacterial strains have been screened to find organisms suitable for biofilm-based biotransformations. This included host strains important for recombinant enzyme expression and strains isolated from biofilters or contaminated soils. Nearly all organisms with bioremediation potential showed good biofilm forming capacities. Pseudomonas sp. strain VLB120DeltaC was chosen as a model organism due to its excellent biofilm forming capacity and its well-studied capability of catalyzing asymmetric epoxidations. A tubular reactor was used for the biotransformation of styrene to (S)-styrene oxide as a model reaction. The process was stable for at least 55 days at a maximal volumetric productivity of 16 g/(L(aq) day) and a yield of 9 mol%. In situ product extraction prevented product inhibition of the catalyst. Biofilm physiology and dynamics are characterized during the biotransformation and limitations and advantages of this reaction concept are discussed.     

Respirometric evaluation and modeling of glucose utilization by Escherichia coli under aerobic and mesophilic cultivation conditions

The study presents a mechanistic model for the evaluation of glucose utilization by Escherichia coli under aerobic and mesophilic growth conditions. In the first step, the experimental data was derived from batch respirometric experiments conducted at 37 degrees C, using two different initial substrate to microorganism (S(0)/X(0)) ratios of 15.0 and 1.3 mgCOD/mgSS. Acetate generation, glycogen formation and oxygen uptake rate profile were monitored together with glucose uptake and biomass increase throughout the experiments. The oxygen uptake rate (OUR) exhibited a typical profile accounting for growth on glucose, acetate and glycogen. No acetate formation (overflow) was detected at low initial S(0)/X(0) ratio. In the second step, the effect of culture history developed under long-term growth limiting conditions on the kinetics of glucose utilization by the same culture was evaluated in a sequencing batch reactor (SBR). The system was operated at cyclic steady state with a constant mean cell residence time of 5 days. The kinetic response of E.coli culture was followed by similar measurements within a complete cycle. Model calibration for the SBR system showed that E. coli culture regulated its growth metabolism by decreasing the maximum growth rate (lower microH) together with an increase of substrate affinity (lower K(S)) as compared to uncontrolled growth conditions. The continuous low rate operation of SBR system induced a significant biochemical substrate storage capability as glycogen in parallel to growth, which persisted throughout the operation. The acetate overflow was observed again as an important mechanism to be accounted for in the evaluation of process kinetics.     

Anaerobic methanethiol degradation in upflow anaerobic sludge bed reactors at high salinity (> or =0.5 M Na(+))

The feasibility of anaerobic methanethiol (MT) degradation at elevated sodium concentrations was investigated in a mesophilic (30 degrees C) lab-scale upflow anaerobic sludge bed (UASB) reactor, inoculated with estuarine sediment originating from the Wadden Sea (The Netherlands). MT was almost completely degraded (>95%) to sulfide, methane and carbon dioxide at volumetric loading rates up to 37 mmol MT x L(-1) x day(-1), 0.5 M sodium (NaCl or NaHCO(3)) and between pH 7.3 and 8.4. Batch experiments revealed that inhibition of MT degradation started at sodium (both NaCl and NaHCO(3)) concentrations exceeding 0.8 M. Sulfide inhibited MT degradation already around 3 mM (pH 8.3).