L. Diels

Enzyme immobilization on/in polymeric membranes: status, challenges and perspectives in biocatalytic membrane reactors (BMRs)

Immobilization of enzymes is beneficial in terms of improving the process economics by enabling enzyme re-use and enhancing overall productivity and robustness. Increasingly, membranes are thought to be good supports for enzyme immobilization. These resulting biocatalytic membranes are integrated in reactors known as biocatalytic membrane reactors (BMRs) which enable the integration of biocatalysis and separation. Often the available commercial membranes require modifications to make them suitable for enzyme immobilization. Different immobilization techniques can be used on such suitable membranes, but no general rules exist for making a choice between them. Despite the advantages of BMR application, there are some issues which need to be addressed in order to achieve up-scaling of such systems. In this review, the different aspects of enzyme immobilization on membranes are discussed to show the complexity of this interdisciplinary technology. In addition, the existing issues which require further investigation are highlighted.

Heavy metals removal by sand filters inoculated with metal sorbing and precipitating bacteria

Abstract Large volumes of wastewater containing metals such as Cd, Zn, Cu, Pb, Hg, Ni or Co are mainly treated by precipitation processes. However, waters treated in such ways do not always meet regulatory standards. And in many cases, ecotaxes must be paid on the heavy metals load in the discharged water. Therefore, a second polishing treatment is often necessary. In order to be economically acceptable, the technology must be cheap and adapted to the treatment of large volumes. The use of sand filters inoculated with heavy metal biosorbing and bioprecipitating bacteria fulfils these objectives. The system is based on a moving bed sand filter. A biofilm is formed on the sand grains after inoculation with heavy metal-resistant bacteria able to biosorb or to bioprecipitate heavy metals. Passage of the wastewater over these biofilms leads to the binding of the metals to the biofilm and consequently the removal of the metals from the wastewater. The metal-laden biofilm is removed from the sand grains in a sand washer created by an airlift for the continuous movement of the filter bed. The metal-loaded biomass is separated from the sand in a labyrinth on the top of the sand washer. Nutrients and a carbon source are provided continuously in the system in order to promote the regrowth of the biofilm on the sand grains. The reactor can be used for the removal of heavy metals, nitrates and some COD. The obtained biosludge contains heavy metals at concentrations of more than 10% of the dry weight. The treatment of the sludge is also taken into account.

The use of bacteria immobilized in tubular membrane reactors for heavy metal recovery and degradation of chlorinated aromatics

Abstract Microbial treatments of waste water can be done in membrane reactors. A membrane installed outside the reactor is used to separate bacteria from the treated effluent. A new membrane reactor concept is presented. The separation membrane is introduced in the reactor and not outside as in a normal one. The membrane plays the role of a separator of two streams and is used at the same time as the immobilizing support for the bacteria. The reactor keeps the bacteria active via a specific nutrient stream that is provided on one side of the membrane. The bacteria grow in and on the membrane where they form an active biofilm. The bacteria can treat the effluent on one side and can be kept active via the nutrient stream at the other side without contamination of the effluent by the nutrient. In this work, the performance of the BICMER (Bacteria Immobilized Composite MEmbrane Reactor) is demonstrated via treatments of effluents containing heavy metals or organic xenobiotics. For heavy metal removal Alcaligenes eutrophus CH34 bacteria were used. These bacteria induce a metal bioprecipitation process that results in the formation of crystalline metal carbonates, which are recovered on a separate column in the reactor. In this way metals can be recovered without disturbing the biofilm on the membrane. Metals such as Cd, Zn, Cu, Pb and Y can be reduced to less than 50 ppb. The metals Co, Ni, Pd and Ge are reduced to below 100 ppb. For organic xenobiotics Alcaligenes eutrophus AE1308 bacteria or other strains (depending on the xenobiotic to be degraded) were used. This strain degrades the xenobiotic 3-chlorobenzoate (Cba) and 2,4-dichlorophenoxyacetic acid to CO2, H2O and chloride). Concentrations of 3 mM Cba could be reduced to less than 0.1 mM. For other toxic organic compounds, different biodegrading strains need to be used.

Benchmark study on algae harvesting with backwashable submerged flat panel membranes.

The feasibility of algae harvesting with submerged flat panel membranes was investigated as pre-concentration step prior to centrifugation. Polishing of the supernatant coming from the centrifuge was evaluated as well. The effect of membrane polymer (polyvinyl chloride [PVC], polyethersulfone polyvinyl-pyrollidone [PES-PVP], poly vinylidene fluoride [PVDF]), pore size (microfiltration [MF], ultrafiltration [UF]), algae cell concentrations and species were investigated at lab-scale. In addition, backwashing as fouling control was compared to standard relaxation. PVDF was the superior polymer, and UF showed better fouling resistance. Backwashing outperformed relaxation in fouling control. The backwashable membranes allowed up to 300% higher fluxes compared to commercial flat panel benchmark (PVC) membranes. Estimations on energy consumption for membrane filtration followed by centrifugation revealed relatively low values of 0.169 kW h/kg of dry weight of algae compared to 0.5 kW h/kg for algae harvesting via classical centrifuge alone.

Treatment of rinsing water from electroless nickel plating with a biologically active moving-bed sand filter

Abstract The MERESAFIN (MEtal REmoval by SAnd Filter INoculation) process presented here was designed to combine the optimum conditions for more than one of the well-known processes of biological metal immobilisation like biosorption and bioprecipitation in a treatment system for industrial waste water. The approach makes use of a continuously operated moving-bed Astrasand® filter which has been inoculated with a mixed population of selected metal biosorbing, bioprecipitating and biodegrading bacteria. One of four pilot plants has been erected at a metal plating company in Vienna (A) to treat waste water from an electroless nickel plating line. In addition to several mg/L of nickel the rinsing water also contains organic acids and inorganic phosphates, which make conventional treatment difficult. Metal laden biomass is continuously removed from the sand grains in the filter and settled in a lamella separator. The thickened biosludge contained 2% of Ni (at only 2–5 mg/L in the feed water), which could be recycled in a shaft furnace. Regeneration of biofilms on the sand is achieved by dosing a cheap carbon source; all other nutrients are available from that specific waste water. For the removal of 0.8 mg/L of nickel the biofilms consumed 8 mg carbon/L and, in addition to 8 mg/L of dissolved oxygen, 3.4 mg NO 3 –N/L as additional electron acceptor. The process was shown to be economically favourable over comparable conventional techniques of metal removal. A further advantage of the biological system is its ability to cleave organo-metal complexes (e.g. nickel lactate in the presented case), to degrade organic molecules like organic acids, surface active substances, etc., often present in industrial waste waters, or to reduce ammonium, nitrite and nitrate. Proposed areas of application comprise the final polishing of industrial and mining water, but also the full treatment of contaminated ground water or drainage water.

Characterization and optimization of β-galactosidase immobilization process on a mixed-matrix membrane.

β-Galactosidase is an important enzyme catalyzing not only the hydrolysis of lactose to the monosaccharides glucose and galactose but also the transgalactosylation reaction to produce galacto-oligosaccharides (GOS). In this study, β-galactosidase was immobilized by adsorption on a mixed-matrix membrane containing zirconium dioxide. The maximum β-galactosidase adsorbed on these membranes was 1.6 g/m², however, maximal activity was achieved at an enzyme concentration of around 0.5 g/m². The tests conducted to investigate the optimal immobilization parameters suggested that higher immobilization can be achieved under extreme parameters (pH and temperature) but the activity was not retained at such extreme operational parameters. The investigations on immobilized enzymes indicated that no real shift occurred in its optimal temperature after immobilization though the activity in case of immobilized enzyme was better retained at lower temperature (5 °C). A shift of 0.5 unit was observed in optimal pH after immobilization (pH 6.5 to 7). Perhaps the most striking results are the kinetic parameters of the immobilized enzyme; while the Michaelis constant (K(m)) value increased almost eight times compared to the free enzyme, the maximum enzyme velocity (V(max)) remained almost constant.

Detailed analysis of petroleum hydrocarbon attenuation in biopiles by high-performance liquid chromatography followed by comprehensive two-dimensional gas chromatography.

Enhanced bioremediation of petroleum hydrocarbons in two biopiles was quantified by high-performance liquid chromatography (HPLC) followed by comprehensive two-dimensional gas chromatography (GCXGC). The attenuation of 34 defined hydrocarbon classes was calculated by HPLC-GCXGC analysis of representative biopile samples at start-up and after 18 weeks of biopile operation. In general, a-cyclic alkanes were most efficiently removed from the biopiles, followed by monoaromatic hydrocarbons. Cycloalkanes and polycyclic aromatic hydrocarbons (PAHs) were more resistant to degradation. A-cyclic biomarkers farnesane, trimethyl-C13, norpristane, pristane and phytane dropped to only about 10% of their initial concentrations. On the other hand, C29-C31 hopane concentrations remained almost unaltered after 18 weeks of biopile operation, confirming their resistance to biodegradation. They are thus reliable indicators to estimate attenuation potential of petroleum hydrocarbons in biopile processed soils.

Laboratory-scale membrane up-concentration and co-anaerobic digestion for energy recovery from sewage and kitchen waste.

This study assessed an alternative concept for co-treatment of sewage and organic kitchen waste in Vietnam. The goal was to apply direct membrane filtration for sewage treatment to generate a permeate that is suitable for discharge. The obtained chemical oxygen demand (COD) concentrations in the permeate of ultrafiltration tests were indeed under the limit value (50 mg/L) of the local municipal discharge standards. The COD of the concentrate was 5.4 times higher than that of the initial feed. These concentrated organics were then co-digested with organic kitchen wastes at an organic loading rate of 2.0 kg VS/m(3).d. The volumetric biogas production of the digester was 1.94 ± 0.34 m(3)/m(3).d. The recovered carbon, in terms of methane gas, accounted for 50% of the total carbon input of the integrated system. Consequently, an electrical production of 64 Wh/capita/d can be obtained when applying the proposed technology with the current wastes generated in Ho Chi Minh City. Thus, it is an approach with great potential in terms of energy recovery and waste treatment.

Removal of nickel from plating rinsing water with a moving-bed sand filter inoculated with metal sorbing and precipitating bacteria

The MERESAFIN (MEtal REmoval by SAnd Filter INoculation) process presented here was designed to combine the optimum conditions for more than one of the well-known processes of biological metal immobilisation like biosorption and bioprecipitation. The approach makes use of a continuously operated moving-bed AstraSand filter which has been inoculated with a mixed population of metal biosorbing and bioprecipitating bacteria. A pilot plant operating at 1 m 3 /h has been erected at a metal plating company in Vienna to treat waste water from an electroless nickel plating line. In addition to several mg/L of nickel the rinsing water also contains some organic acids and inorganic phosphates, which make conventional treatment difficult. The main laboratory experiments as well as preliminary results from the pilot installation are presented.

Microbial fixation of CO2 in water bodies and in drylands to combat climate change, soil loss and desertification

The growing concern for the increase of the global warming effects due to anthropogenic activities raises the challenge of finding novel technological approaches to stabilize CO2 emissions in the atmosphere and counteract impinging interconnected issues such as desertification and loss of biodiversity. Biological-CO2 mitigation, triggered through biological fixation, is considered a promising and eco-sustainable method, mostly owing to its downstream benefits that can be exploited. Microorganisms such as cyanobacteria, green algae and some autotrophic bacteria could potentially fix CO2 more efficiently than higher plants, due to their faster growth. Some examples of the potential of biological-CO2 mitigation are reported and discussed in this paper. In arid and semiarid environments, soil carbon sequestration (CO2 fixation) by cyanobacteria and biological soil crusts is considered an eco-friendly and natural process to increase soil C content and a viable pathway to soil restoration after one disturbance event. Another way for biological-CO2 mitigation intensively studied in the last few years is related to the possibility to perform carbon dioxide sequestration using microalgae, obtaining at the same time bioproducts of industrial interest. Another possibility under study is the exploitation of specific chemotrophic bacteria, such as Ralstonia eutropha (or picketii) and related organisms, for CO2 fixation coupled with the production chemicals such as polyhydroxyalkanoates (PHAs). In spite of the potential of these processes, multiple factors still have to be optimized for maximum rate of CO2 fixation by these microorganisms. The optimization of culture conditions, including the optimal concentration of CO2 in the provided gas, the use of metabolic engineering and of dual purpose systems for the treatment of wastewater and production of biofuels and high value products within a biorefinery concept, the design of photobioreactors in the case of phototrophs are some of the issues that, among others, have to be addressed and tested for cost-effective CO2 sequestration.