Performance of a scaled-up Microbial Fuel Cell with iron reduction as the cathode reaction

Scale-up studies of Microbial Fuel Cells are required before practical application comes into sight. We studied an MFC with a surface area of 0.5 m² and a volume of 5 L. Ferric iron (Fe³⁺) was used as the electron acceptor to improve cathode performance. MFC performance increased in time as a combined result of microbial growth at the bio-anode, increase in iron concentration from 1 g L⁻¹ to 6 g L⁻¹, and increased activity of the iron oxidizers to regenerate ferric iron. Finally, a power density of 2.0 W m⁻² (200 W m⁻³) was obtained. Analysis of internal resistances showed that anode resistance decreased from 109 to 7 mΩ m², while cathode resistance decreased from 939 to 85 mΩ m². The cathode was the main limiting factor, contributing to 58% of the total internal resistance. Maximum energy efficiency of the MFC was 41%.     

Site-directed mutagenesis and X-ray crystallography of two phospholipase A2 mutants: Y52F and Y73F

Tyr52 and Tyr73 are conserved amino acid residues throughoutall vertebrate phospholipases A₂. They are part of an extendedhydrogen bonding system that links the N-terminal -NH⁺₃ -groupto the catalytic residues His48 and Asp99. These tyrosines werereplaced by phenylalanines in a porcine pancreatic phospholipaseA₂ mutant, in which residues 62–66 had been deleted (62–66PLA₂).The mutations did not affect the catalytic properties of theenzyme, nor the folding kinetics. The stability against denaturatlonby guanidine hydrochloride was decreased, however. To analysehow the enzyme compensates for the loss of the tyrosine hydroxylgroup, the X-ray structures of the Y52F and AY73F mutants weredetermined. After crystallographic refinement the final crystallographicR-factors were 18.1% for the %Y52F mutant (data between 7 and2.3 Å resolution) and 19.1% for the Y73F mutant (databetween 7 and 2.4 Å resolution). No conformational changesoccurred in the mutants compared with the 62–66PLA₂, butan empty cavity formed at the site of the hydroxyl group ofthe former tyrosine. In both mutants the Asp99 side chain losesone of its hydrogen bonds and this might explain the observeddestabilization.     

Incorporation of an unnatural amino acid in the active site of porcine pancreatic phospholipase A2. Substitution of histidine by l,2,4-triazole-3-alanine yields an enzyme with high activity at acidic pH

The effect of the substitution of the active site histidine48 by the unnatural 1,2,4-triazole-3-alanine (TAA) amino acidanalogue in porcine pancreas phospholipase A₂ (PLA₂) was studied.TAA was introduced biosynthetically using a his-auxotrophicEscherichia coli strain. To study solely the effect of the substitutionof the active site histidine, two nonessential histidines (i.e.His17 and His 115) were replaced by asparagines, resulting ina fully active mutant enzyme (His-PLA₂). In this His-PLA₂ thesingle histidine at position 48 was substituted by TAA withan incorporation efficiency of about 90%, giving a mixture ofHis-PLA₂ and TAA-PLA₂. Based on the charge difference at acidicpH, both forms could be separated by FPLC, allowing for thepurification of TAA-PLA₂ free from His-PLA₂. At pH 6, TAA-PLA₂has a fivefold reduced activity compared with His-PLA₂. Thisreduced activity paralells a reduced rate of covalent modificationwith p-nitrophenacyl bromide of TAA-PLA₂ compared with His-PLA₂.Competitive inhibition gave comparable IC₅₀ values for WT-PLA₂,His-PLA₂ and TAA-PLA₂. These results indicate that the reductionin activity is not caused by a different affinity for the substrate,but more likely results from a reduced k_cat value in TAA-PLA₂.The enzymatic activities for native and mutant PLA₂s were measuredat different pH values. For WT-PLA₂ and His-PLA₂ the activityis optimal at pH 6 and is strongly deminished at acidic pH,with no observable activity at pH 3. In contrast, TAA-PLA₂ isas active at pH 3 as at pH 6. Most likely, the decrease in activityobserved for WT-PLA₂ and His-PLA₂ is caused by the protonationof the active site His48, which is the general base involvedin the activation of the nucleophilic water molecule. In TAA-PLA₂,however, the active site residue TAA48 is unprotonated at bothpH 3 and 6 as a result of the low pK_a of TAA compared with histidine.     

Hypadapter: An adaptive hypertext system for exploratory learning and programming

We have developed an adaptive hypertext system designed to individually support exploratory learning and programming activities in the domain of Common Lisp. Endowed with domain-specific knowledge represented in a hyperspace of topics, the system builds up a detailed model of the user's expertise which it utilizes to provide personalized assistance. Unlike other work emerging in the field of adaptive hypertext systems, our approach exploits domain and user modelling techniques to support individuals in different ways. The system not only generates individualized presentations of topic nodes, but also provides adaptive navigational assistance for link-based browsing. By identifying and suggesting useful hyperlinks according to the user's knowledge state and preferences, the system encourages and guides exploration. While browsing through the hyperspace of topics, the system analyses the user's navigational behaviour to infer the user's learning progress and to dynamically adapt presentations of topics and links accordingly.     

Einfluß der Unterbrechergeometrie auf die Arbeitsübertragung und den Wirkungsgrad von Seitenkanalverdichtern

Zusammenfassung Zur Trennung von Druck- und Saugseite des Seitenkanals ist ein Unterbrecher erforderlich, der das direkte überstr?men des verdichteten Gases verhindert. Der Unterbrecher bewirkt aber nicht nur die Abdichtung, sondern er beeinflu?t auch wesentlich den Expansionsvorgang des überstr?menden Unterbrechermassestromes und damit wiederum den Verdichtungsvorgang im Seitenkanal. Um den Einflu? der Unterbrechergeometrie auf diese Vorg?nge zu ermitteln, wurden unterschiedliche Unterbrechermodule in einem Seitenkanalverdichter bei Druck- und Vakuumbetrieb untersucht. Der Umfangswinkel des Unterbrechers und die Oberfl?chengeometrie beeinflussen den Expansionsvorgang und den polytropen Kupplungswirkungsgrad unterschiedlich im Bereich von η=0,41 bis 0,49. Damit ist aufgezeigt, da? eine Optimierung des Unterbrechers notwendig ist, um m?glichst adiabate Zustands?nderungen im Seitenkanalverdichter zu erreichen. Der relative Unterbrecherwinkel soll für Schaufelzahlen von 40 bis 60 α_U/2π=0,115 bis 0,135 betragen.     

Polychlorierte Biphenyle (PCB) in Fischen aus Teichwirtschaften und Nachklärteichen

European Food Research and Technology - Das Muskelgewebe von 58 Fischen aus Teichwirtschaften und 17 Fischen aus Nachklärteichen wurde auf seinen Gehalt an den sechs in der...     

Alcohol production through volatile fatty acids reduction with hydrogen as electron donor by mixed cultures

In this research we demonstrated a new method to produce alcohols. It was experimentally feasible to produce ethanol, propanol and butanol from solely volatile fatty acids (VFAs) with hydrogen as electron donor. In batch tests, VFAs such as acetic, propionic and butyric acids were reduced by mixed microbial cultures with a headspace of 1.5 bar of hydrogen. Observed alcohol concentrations were 3.69 ± 0.25 mM of ethanol, 8.08 ± 0.85 mM of propanol and 3.66 ± 0.05 mM of n-butanol. The conversion efficiency based on the electron balance was 55.1 ± 5.6% with acetate as substrate, 50.3 ± 4.7% with propionate and 46.7 ± 2.2% with n-butyrate. Methane was the most predominant by-product in each batch experiment, 33.6 ± 9.6% of VFA and hydrogen was converted to methane with acetate as substrate; which was 27.1 ± 7.1% with propionate and 36.6 ± 2.2% with n-butyrate. This VFAs reducing renewable fuel production process does not require carbohydrates like fermentable sugars, but uses biomass with high water content or low sugar content that is unsuitable as feedstock for current fermentation processes. This so-called low-grade biomass is abundantly present in many agricultural areas and is economically very attractive feedstock for the production of biofuels.     

Hydrogen production with a microbial biocathode

This paper, for the first time, describes the development of a microbial biocathode for hydrogen production that is based on a naturally selected mixed culture of electrochemically active micro-organisms. This is achieved through a three-phase biocathode startup procedure that effectively turned an acetate- and hydrogen-oxidizing bioanode into a hydrogen-producing biocathode by reversing the polarity of the electrode. The microbial biocathode that was obtained in this way had a current density of about -1.2 A/Nm2 at a potential of -0.7 V. This was 3.6 times higher than that of a control electrode (-0.3 A/m2). Furthermore, the microbial biocathode produced about 0.63 m3 H2/m3 cathode liquid volume/day at a cathodic hydrogen efficiency of 49% during hydrogen yield tests, whereas the control electrode produced 0.08 m3 H2/m3 cathode liquid volume/day at a cathodic hydrogen efficiency of 25%. The effluent of the biocathode chamber could be used to inoculate another electrochemical cell that subsequently also developed an identical hydrogen-producing biocathode (-1.1 A/m2 at a potential of -0.7 V). Scanning electron micrographs of both microbial biocathodes showed a well-developed biofilm on the electrode surface.     

Function of the fully conserved residues Asp99, Tyr52 and Tyr73 in phospholipase A2

In the active centre of pancreatic phospholipase A₂ His48 isat hydrogen-bonding distance to Asp99. This Asp-His couple isassumed to act together with a water molecule as a catalytictriad. Asp99 is also linked via an extended hydrogen bondingsystem to the side chains of Tyr52 and Tyr73. To probe the functionof the fully conserved Asp99, Tyr52 and Tyr73 residues in phospholipaseA₂, the Asp99 residue was replaced by Asn, and each of the twotyrosines was separately replaced by either a Phe or a Gln.The catalytic and binding properties of the Phe52 and Phe73mutants did not change significantly relative to the wild-typeenzyme. This rules out the possibility that either one of thetwo Tyr residues in the wild-type enzyme can function as anacyl acceptor or proton donor in catalysis. The Gln73 mutantcould not be obtained in any significant amounts probably dueto incorrect folding. The Gln52 mutant was isolated in low yield.This mutant showed a large decrease in catalytic activity whileits substrate binding was nearly unchanged. The results suggesta structural role rather than a catalytic function of Tyr52and Tyr73. Substitution of asparagine for aspartate hardly affectsthe binding constants for both monomeric and micellar substrateanalogues. Kinetic characterization revealed that the Asn99mutant has retained no less than 65% of its enzymatic activityon the monomeric substrate rac 1,2-dihexanoyldithio-propyl-3-phosphocholine,probably due to the fact that during hydrolysis of monomericsubstrate by phospholipase A₂ proton transfer is not the rate-limitingstep. The Asp to Asn substitution decreases the catalytic rateon micellar 1,2-dioctanoyl-sn-glycero-3-phosphocholine 25-fold.To explain this remaining activity we suggest that in the mutantthe Asn99 orients His48 in the same way as Asp99 orients His48in native phospholipase A₂ and that the lowered activity iscaused by a reduced stabilization of the transition state.