BioCatNet: A Database System for the Integration of Enzyme Sequences and Biocatalytic Experiments

The development of novel enzymes for biocatalytic processes requires knowledge on substrate profile and selectivity; this can be derived from databases and from publications. Often, these sources lack time‐course data for the substrate or product, and an unambiguous link between experiment and enzyme sequence. The lack of integrated, original data hampers the comprehensive analysis of enzyme kinetics and the evaluation of sequence–function relationships. In order to accelerate enzyme engineering, BioCatNet integrates protein sequence, protein structure, and experimental data for a given enzyme family. BioCatNet explicitly assigns the enzyme sequence to the experimental data, which consists of information on reaction conditions and time‐course data. BioCatNet facilitates the consistent documentation of reaction conditions, the archiving of time‐course data, and the efficient exchange of experimental data among collaborators. Data integration is demonstrated for three case studies by using the TEED (Thiamine diphosphate‐dependent Enzymes Engineering Database).     

Recovery of mineral fertiliser N and slurry N in continuous silage maize using the <Superscript>15</Superscript>N and difference methods

The stable isotope technique and the difference method are common approaches for estimating fertiliser N uptake efficiency. Both methods, however, have limitations and their suitability may depend on N management and environmental conditions. A field experiment was conducted on a humus sandy soil in northern Germany to estimate fertiliser N uptake efficiency of silage maize in the year of application (Zea mays L.) by the stable isotope and the difference method as influenced by the type of N fertiliser (mineral vs. cattle slurry), the application mode (separate or combined application), and N rate. Seven N treatments were included (0, 50, 100 and 150 kg mineral N ha⁻¹; 20, 40 m3 cattle slurry ha⁻¹; 50 kg mineral N ha⁻¹ plus 40 m3 slurry ha⁻¹), where either mineral N or slurry N was labelled, and mineral N was split into two dressings. In addition, 4.1 kg ha⁻¹ labelled mineral N was incorporated into otherwise unlabelled treatments (0, 20, 40 m3 ha⁻¹, and 50 kg mineral N ha⁻¹ plus 40 m3 ha⁻¹) to estimate N uptake from the upper soil layer. Uptake of ¹⁵N was followed in leaves, stalk, ear, and the whole crop. Fertiliser N uptake efficiency (FNUE_15N) of mineral fertiliser N obtained by the isotope technique ranged between 51 and 61%. Recovered fertiliser N was mainly found in the ear, while less labelled N remained in leaves and the stalk. The nitrogen rate tended to increase the amount of recovered N, but the effect was not consistent among plant parts and the whole crop. Plant N uptake from non-fertiliser N was found to increase N input up to 100 kg N ha⁻¹. Nitrogen recoveries of the two mineral N dressings were similar for the different plant parts as well as for the whole crop. Fertiliser N uptake efficiency (FNUE_diff) of mineral N estimated by the difference method resulted in substantially higher values compared to FNUE_15N, varying between 56 and 98%. More N was taken up from the upper soil layer with increasing N supply, which is regarded as a major error source of the difference method. Slurry N was taken up less efficient in the year of application than mineral fertiliser N as indicated by recovery rates of 21–22% (FNUE_15N) and 39–62% (FNUE_diff), respectively. When mineral N and slurry were applied together, the difference method estimated significantly lower N uptake efficiencies for both mineral and slurry N compared to a single application, while values obtained by the isotope method were not affected.     

Hydrogeochemische Hintergrundwerte der Grundw?sser Deutschlands als Web Map Service

One of the main objectives of the EC water framework directive is to ensure good chemical status for all groundwater bodies. For this reason the geological surveys of Germany have produced a nationwide map of the background values of groundwater. Only naturally occurring inorganic parameters were taken into account, including the relevant major and trace elements. Based on the hydrogeological map of Germany at the scale of 1:200,000 (HüK 200), and its delimited hydrogeological regions, a total of 186 hydrogeochemical units were defined and mapped geochemically. This involved collection of more than 52,000 groundwater samples in a database and allocation to their appropriate hydrogeochemical units. In order to separate anomalies within the data-sets from the underlying normal population, probability nets were used as a statistical tool. The procedure allowed to determine the normal populations of the investigated parameters within the hydrogeochemical units and to quantify them in the form of percentiles. The resulting hydrogeochemical background values are accessible through an internet web map service that includes an online map application. The main intention of this technical note is to inform the reader about the existing internet service, which necessitates some web-site content duplication.     

Optimization of adsorptive immobilization of alcohol dehydrogenases

In this work, a systematic examination of various parameters of adsorptive immobilization of alcohol dehydrogenases (ADHs) on solid support is performed and the impact of these parameters on immobilization efficiency is studied. Depending on the source of the enzymes, these parameters differently influence the immobilization efficiency, expressed in terms of residual activity and protein loading. Residual activity of 79% was achieved with ADH from bakers' yeast (YADH) after optimizing the immobilization parameters. A step-wise drying process has been found to be more effective than one-step drying. A hypothesis of deactivation through bubble nucleation during drying of the enzyme/glass bead suspension at low drying pressure (<45 kPa) is experimentally verified. In the case of ADH from Lactobacillus brevis (LBADH), >300% residual activity was found after drying. Hyperactivation of the enzyme is probably caused by structural changes in the enzyme molecule during the drying process. ADH from Thermoanaerobacter species (ADH T) is found to be stable under drying conditions (>15 kPa) in contrast to LBADH and YADH.     

Steganographie und Wasserzeichen – Aktueller Stand und neue Herausforderungen

Zusammenfassung Digitale Mediendaten (z. B. Bild-, Ton-, Videodaten oder aber auch symbolische Repr?sentationen) haben in den letzten Jahren stark an Bedeutung gewonnen. Sie ?ffnen neue M?rkte und Anwendungsfelder, erfordern aber gleichzeitig auch neue Sicherheitsmechanismen zum Schutz der Mediendaten, zur Identifikation oder Authentisierung in verschiedenen Repr?sentationsformen bis hin zum Schutz urheberrechtlicher Ansprüche.     

纤维素纤维及其混纺机织物的连续前处理

描述了德国Zschimmer＆Schwarz公司开发的机织物前处理系统Optasize和Optasteam。，结果表明：用Optasize／Optasteam工艺可应用在日常生产中，并为定制加工提供解决方案以满足客户要求，其流程简洁经济而且可靠性高。通过采用这一工艺最初不同起因的织物质量差异可由此拉平。     

"Soft" metallic contact to isolated C60 molecules

C60 adsorbed on a monolayer of hexaazatriphenylene-hexanitrile (HATCN) on Ag(111) is investigated by ultraviolet photoelectron spectroscopy (UPS) and scanning tunneling microscopy. UPS and quantum-mechanical modeling show that HATCN chemisorbed on Ag(111) displays metallic character. This metallic molecular layer decouples C60 electronically from the Ag substrate and simultaneously acts both as template for the stable adsorption of isolated C60 molecules at room temperature and as "soft" metallic contact for subsequently deposited molecules.     

DNA-oligonucleotide encapsulating liposomes as a secondary signal amplification means

A novel liposome-based signal amplification system was developed by encapsulating DNA oligonucleotides within antibody-tagged liposomes and subsequently detecting the oligonucleotide with dye-encapsulating liposomes for double signal enhancement. In this sandwich immunoassay, the model analyte, protective antigen protein from B. anthracis, was captured by one set of antibodies immobilized in microtiter plate wells and detected using a second antibody conjugated to oligonucleotide-encapsulating liposomes. Bound liposomes were lysed releasing the encapsulated fluorescein-tagged DNA 25-mer probe, which was then permitted to hybridize with its complementary sequence immobilized in a second plate. Finally, the amount of oligonucleotide was detected through the addition of anti-fluorescein antibody tagged dye-encapsulating liposomes. These secondary liposomes allowed for a approximately 400x lower LOD than detection of the fluorescein-labeled probe alone. Several aspects were investigated, including the encapsulation of various oligonucleotide concentrations within liposomes; oligonucleotide hybridization times and buffers; degree of anti-fluorescein antibody coverage on the liposomes; and immobilized anti-protective antigen antibody concentration. We found that the encapsulation efficiency increased with the starting oligonucleotide concentration. As many as 4000 DNA 25-mers were successfully entrapped in the liposome, and minimal leakage was observed over the course of 8 months. When used in the sandwich immunoassay, a limit of detection of 4.1 ng/mL protective antigen was observed with an upper limit of 5000 ng/mL. Due to the endless combination of DNA oligonucleotide sequences, this assay lends itself perfectly for multiplexing on the order of tens to hundreds of analytes.     

Application of ganglioside-sensitized liposomes in a flow injection immunoanalytical system for the determination of cholera toxin

Cholera, an acute infectious disease associated with water and seafood contamination, is caused by the bacterium Vibrio cholerae, which lives and colonizes in the small intestine and secretes cholera toxin (CT), a causative agent for diarrhea in humans. Based on earlier lateral flow assays, a flow injection liposome immunoanalysis (FILIA) system with excellent sensitivity was developed in this study for the determination of CT at zeptomole levels. Ganglioside (GM1), found to have specific affinity toward CT, was inserted into the phospholipid bilayer during the liposome synthesis. These GM1-sensitized, sulforhodamine B (SRB) dye-entrapping liposomes were used as probes in the FILIA system. Anti-CT antibodies were immobilized in its microcapillary. CT was detected by the formation of a sandwich complex between the immobilized antibody and GM1 liposomes. During the assay, the sample was introduced first into the column, and then liposomes were injected to bind to all CT captured by the antibody in the microcapillary. Subsequently, the SRB dye molecules were released from the bound liposomes via the addition of the detergent octyl glucopyranoside. The released dye molecules were transported to a flow-through fluorescence detector for quantification. The FILIA system was optimized with respect to flow rate, antibody concentration, liposome concentration, and injected sample volume. The calibration curve for CT had a linear range of 10-16 to 10-14 g mL-1. The detection limit of this immunosensor was 6.6 x 10(-17) g mL-1 in 200-microL samples (equivalent to 13 ag or 1.1 zmol).