Heleen De Wever

Biodiversity and population dynamics of microorganisms in a full-scale membrane bioreactor for municipal wastewater treatment

The total, ammonia-oxidizing, and denitrifying Bacteria in a full-scale membrane bioreactor (MBR) were evaluated monthly for over one year. Microbial communities were analyzed by denaturing gradient gel electrophoresis (DGGE) and clone library analysis of the 16S rRNA and ammonia monooxygenase (amoA) and nitrous oxide reductase (nosZ) genes. The community fingerprints obtained were compared to those from a conventional activated sludge (CAS) process running in parallel treating the same domestic wastewater. Distinct DGGE profiles for all three molecular markers were observed between the two treatment systems, indicating the selection of specific bacterial populations by the contrasting environmental and operational conditions. Comparative 16S rRNA sequencing indicated a diverse bacterial community in the MBR, with phylotypes from the α- and β-Proteobacteria and Bacteroidetes dominating the gene library. The vast majority of sequences retrieved were not closely related to classified organisms or displayed relatively low levels of similarity with any known 16S rRNA gene sequences and thus represent organisms that constitute new taxa. Similarly, the majority of the recovered nosZ sequences were novel and only moderately related to known denitrifiers from the α- and β-Proteobacteria. In contrast, analysis of the amoA gene showed a remarkably simple ammonia-oxidizing community with the detected members almost exclusively affiliated with the Nitrosomonas oligotropha lineage. Major shifts in total bacteria and denitrifying community were detected and these were associated with change in the external carbon added for denitrification enhancement. In spite of this, the MBR was able to maintain a stable process performance during that period. These results significantly expand our knowledge of the biodiversity and population dynamics of microorganisms in MBRs for wastewater treatment.

Integrated bioprocess for long-term continuous cultivation of Clostridium acetobutylicum coupled to pervaporation with PDMS composite membranes.

A continuous cultivation of Clostridium acetobutylicum ATCC 824 is described using a two-stage design to mimic the two phases of batch culture growth of the organism. A hydrophobic pervaporation unit was coupled to the second fermentor containing the highest solvent titers. This in situ product recovery technology efficiently decreased butanol toxicity in the fermentor while the permeate was enriched to 57-195 g L(-1) total solvents depending on the solvent concentrations in the fermentor. By the alleviation of product inhibition, the glucose concentration could be increased from 60 to 126 g L(-1) while the productivity increased concomitantly from 0.13 to 0.30 g L(-1)h(-1). The continuous fermentation was conducted for 1172 h during which the pervaporation was coupled to the second fermentor for 475 h with an average flux of 367 g m(-2)h(-1). The energy consumption was calculated for a 2 wt.% n-butanol fermentation broth and compared with the conventional process.

Valorization of Cereal Based Biorefinery Byproducts:Reality and Expectations

The growth of the biobased economy will lead to an increase in new biorefinery activities. All biorefineries face the regular challenges of efficiently and economically treating their effluent to be compatible with local discharge requirements and to minimize net water consumption. The amount of wastes resulting from biorefineries industry is exponentially growing. The valorization of such wastes has drawn considerable attention with respect to resources with an observable economic and environmental concern. This has been a promising field which shows great prospective toward byproduct usage and increasing value obtained from the biorefinery. However, full-scale realization of biorefinery wastes valorization is not straightforward because several microbiological, technological and economic challenges need to be resolved. In this review we considered valorization options for cereals based biorefineries wastes while identifying their challenges and exploring the opportunities for future process.

Electro-Fermentation – Merging Electrochemistry with Fermentation in Industrial Applications

Electro-fermentation (EF) merges traditional industrial fermentation with electrochemistry. An imposed electrical field influences the fermentation environment and microbial metabolism in either a reductive or oxidative manner. The benefit of this approach is to produce target biochemicals with improved selectivity, increase carbon efficiency, limit the use of additives for redox balance or pH control, enhance microbial growth, or in some cases enhance product recovery. We discuss the principles of electrically driven fermentations and how EF can be used to steer both pure culture and microbiota-based fermentations. An overview is given on which advantages EF may bring to both existing and innovative industrial fermentation processes, and which doors might be opened in waste biomass utilization towards added-value biorefineries.

Integrated conversion of food waste diluted with sewage into volatile fatty acids through fermentation and electricity through a fuel cell

In this study, domestic wastewater was given a second life as dilution medium for concentrated organic waste streams, in particular artificial food waste. A two-step continuous process with first volatile fatty acid (VFA)/hydrogen production and second electricity production in microbial fuel cells (MFCs) was employed. For primary treatment, bioreactors were optimized to produce hydrogen and VFAs. Hydrolysis of the solids and formation of fermentation products and hydrogen was monitored. In the second step, MFCs were operated batch-wise using the effluent rich in VFAs specifically acetic acid from the continuous reactor of the first step. The combined system was able to reduce the chemical oxygen demand load by 90%. The concentration of VFAs was also monitored regularly in the MFCs and showed a decreasing trend over time. Further, the anode potential changed from -500 to 0 mV vs. Ag/AgCl when the VFAs (especially acetate) were depleted in the system. On feeding the system again with the effluent, the anode potential recovered back to -500 mV vs. Ag/AgCl. Thus, the overall aim of converting chemical energy into electrical energy was achieved with a columbic efficiency of 46% generating 65.33 mA/m2 at a specific cell potential of 148 mV.

Advances in in-situ product recovery (ISPR) in whole cell biotechnology during the last decade.

The review presents the state-of-the-art in the applications of in-situ product recovery (ISPR) in whole-cell biotechnology over the last 10years. It summarizes various ISPR-integrated fermentation processes for the production of a wide spectrum of bio-based products. A critical assessment of the performance of various ISPR concepts with respect to the degree of product enrichment, improved productivity, reduced process flows and increased yields is provided. Requirements to allow a successful industrial implementation of ISPR are also discussed. Finally, supporting technologies such as online monitoring, mathematical modeling and use of recombinant microorganisms with ISPR are presented.

Pervaporative recovery of ABE during continuous cultivation: enhancement of performance.

Acetone, butanol and ethanol were produced in a continuous two-stage fermentation integrated with pervaporation using freely suspended cells of C. acetobutylicum ATCC 824. PDMS composite pervaporation membranes were directly coupled to the second fermentor which lead to decreased solvent titers. Overall productivity was increased from 0.45 g L(-1) h(-1) to 0.88 g L(-1) h(-1) when increasing the carbohydrate concentration in the feed from 60 to 120 g L(-1). The highest overall productivity of 1.13 g L(-1) h(-1) was achieved when increasing the carbohydrate concentration further to 150 g L(-1) even though productivity decreased significantly in the first fermentor due to substrate inhibition. In this phase that lasted 200 h, the average flux reached 0.621 kg m(-2) h(-1) and the total solvent concentration in the permeate was 202 g L(-1). High solvent titers in the second fermentor were beneficial for the performance of the pervaporation unit leading to higher fluxes and total solvent concentrations in the permeate.

Metabolism of 2-Mercaptobenzothiazole by Rhodococcus rhodochrous

ABSTRACT 2-Mercaptobenzothiazole, which is mainly used in the rubber industry as a vulcanization accelerator, is very toxic and is considered to be recalcitrant. We show here for the first time that it can be biotransformed and partially mineralized by a pure-culture bacterial strain of Rhodococcus rhodochrous. Three metabolites, among four detected, were identified.

Validation of a supervisory control system for energy savings in membrane bioreactors

The application of fixed operational protocols and settings for membrane bioreactors (MBR) often leads to suboptimal filtration conditions due to the dynamic nature of mixed liquor characteristics. With regard to process optimization and energy savings, the potential benefits of a dynamic control system, enabling to adapt fouling control actions (ACS outputs) in an automated way to the actual mixed liquor fouling propensity, are thus obvious. In this paper, the pilot-scale validation of such an advanced control system (ACS) is elaborated. A specific on-line fouling measurement method, the MBR-VFM (VITO Fouling Measurement), was used for the evaluation of the mixed liquors reversible fouling propensity, which was used as a primary ACS input parameter. A first series of tests with a gradual increase in complexity of the selected input and output parameters indicated the functionality of the ACS and demonstrated a substantial reduction of aeration, however sometimes at the expense of a higher fouling rate. The ACS was further fine-tuned and subsequently tested for a longer period under more dynamic operating conditions. A significant correlation was found between the reversible fouling potential measured by the MBR-VFM and the on-line permeability, indicating that the MBR-VFM is a suitable ACS input. Furthermore, an average 22% reduction in aeration flow to the membranes could be achieved without any obvious negative effect on filtration performance. This indicates that this approach is promising to optimize energy consumption in MBRs.

Modeling pure culture heterotrophic production of polyhydroxybutyrate (PHB)

In this contribution a mechanistic model describing the production of polyhydroxybutyrate (PHB) through pure-culture fermentation was developed, calibrated and validated for two different substrates, namely glucose and waste glycerol. In both cases, non-growth-associated PHB production was triggered by applying nitrogen limitation. The occurrence of some growth-associated PHB production besides non-growth-associated PHB production was demonstrated, although it is inhibited in the presence of nitrogen. Other phenomena observed experimentally and described by the model included biomass growth on PHB and non-linear product inhibition of PHB production. The accumulated impurities from the waste substrate negatively affected the obtained maximum PHB content. Overall, the developed mathematical model provided an accurate prediction of the dynamic behavior of heterotrophic biomass growth and PHB production in a two-phase pure culture system.