Characterization of a heterodimeric GH2 β-galactosidase from Lactobacillus sakei Lb790 and formation of prebiotic galacto-oligosaccharides

The lacLM genes from Lactobacillus sakei Lb790, encoding a heterodimeric β-galactosidase that belongs to glycoside hydrolase family GH2, were cloned and heterologously expressed in Escherichia coli . Subsequently, the recombinant β-galactosidase LacLM was purified to apparent homogeneity and characterized. The enzyme is a β-galactosidase with narrow substrate specificity because o-nitrophenyl-β-D-galactopyranoside (oNPG) was efficiently hydrolyzed, whereas various structurally related oNP analogues were not. The K(m) and k(cat) values for oNPG and lactose were 0.6 mM and 180 s(-1) and 20 mM and 43 s(-1), respectively. The enzyme is inhibited competitively by its two end-products D-galactose and D-glucose (K(i) values of 180 and 475 mM, respectively). As judged by the ratio of the inhibition constant to the Michaelis constant, K(i)/K(m), this inhibition is only very moderate and much less pronounced than for other microbial β-galactosidases. β-Galactosidase from L. sakei possesses high transgalactosylation activity and was used for the synthesis of galacto-oligosaccharides (GalOS), employing lactose at a concentration of 215 g/L. The maximum GalOS yield was 41% (w/w) of total sugars at 77% lactose conversion and contained mainly non-lactose disaccharides, trisaccharides, and tetrasaccharides with approximately 38, 57, and 5% of total GalOS formed, respectively. The enzyme showed a strong preference for the formation of β-(1→6)-linked transgalactosylation products, whereas β-(1→3)-linked compounds were formed to a lesser extent and β-(1→4)-linked reaction products could not be detected.     

A Quantum Statistical Approach to Remediation Effect of Humic Substances

Among various remediation factors, dissolved organic matter including humic substances (HS) has substantial effect on environmental contamination significantly changing the contaminant??s degradation, bioavailability, reactivity, and immobilization. However, the effects strongly depend on HS concentrations and their aromaticity index (AI). To understand underlying phenomena of remediation action of HS, which is revealed to occur within a definite interval of HS concentrations in water solution, a quantum statistical approach is supposed. Developing this approach, a model of protons as Fermi particles in humic substances was advanced for the first time and applied to describe transformations of HS molecules, i.e., multipoles into micelle structures, which in turn provide for mediating effects in water. Sufficiently high concentration of micelle granules in water solution exists if the concentration of HS lies within a definite interval. It was demonstrated applying a grand canonical Gibbs distribution method to a statistical ensemble of HS particles. Our approach allows for understanding and quantifying some biological and physiological processes connected with mediating action of HS, as for example the reversible red cell aggregation influenced by HS, adsorption of HS particles by cancer cells, and effect of HS on human resistibility to inflammatory processes of different kinds. Application of our results to water systems may be helpful to optimize waste processing and disposal.     

Salinity stress in tomatoes can be alleviated by grafting and potassium depending on the rootstock and K-concentration employed

Plant production under salinity requires increased capacity for K⁺ homeostasis. For this purpose, supplementary K₂SO₄ in the nutrient solution and grafting on a tolerant rootstock were employed in two experiments to test whether grafting, potassium and their interactions can alleviate salinity stress in tomato (Solanum lycopersicum L.). In Exp-ion, plants were cultivated for 122 days to compare different ionic compositions: EC 9 dS m⁻¹ in EC_all (by macro-nutrients) and in EC_NaCl (by 64.2 mM NaCl), EC 12 dS m⁻¹ in EC_K (EC_NaCl + 25.8 mM K⁺). Exp-K⁺ was established to compare K⁺ concentrations of 6, 16 and 36 mM at 150 mM NaCl. In both Experiments, ‘ZS-5’, selected as a salt sensitive cultivar, was either self-grafted or grafted onto the cultivar ‘Edkawi’, reported as salt tolerant. Yield and growth, minerals, gas exchange, soluble sugars, and proline were analyzed. Different ionic treatments affected almost all characteristics considered while differences between rootstocks were rarely observed. No pronounced differences were found in shoot growth, yield and gas exchange between EC_all and EC_NaCl. EC_K did not show any salinity alleviative effects but inhibited even growth compared with the other treatments. In Exp-K⁺, 16 mM K⁺ increased plant growth, leaf soluble sugars and proline concentrations. 36 mM K⁺ did not further reduce upper leaf Na⁺ although leaf K⁺ concentration increased significantly. The results indicated that the response of tomato plant to NaCl stress was principally attributed to the osmotic component in Exp-ion, excessive K⁺ showed no mitigating effect on fruit yield and shoot growth. However, 16 mM K⁺ in the root environment enhanced the salt adaptive capacity of plants stressed at 150 mM NaCl. The use of the tolerant rootstock resulted in no ameliorative effects, owing to its susceptibility to blossom-end rot, failure in enhancing photosynthesis, and ineffectiveness of restraining the long-distance transport of Na⁺.     

Beziehung zwischen Bodenorganismen und Gefügeentwicklung auf Löß-Neuland unter Acker- und Waldnutzung

Relationship between soil organisms and soil structure in new reclaimed soils derived from loess under agricultural and forestral use In the Rhenish Browncoal District est of Cologne he question occurs wether forestry or agriculture is the better form of primary use to force structure development in new reclaimed soils derived from loess. The investigations on 10–25 years old reclaimed soils under forestral and agricultural use indicate that humus content, microbiological activities as well as the number of soil meso- and macrofauna individuals like earthworms are increasing with growing age of the forest soils. These processes and the strucutre development are faster under forestral use. Here after about 20–30 years conditions are similiar to those of undisturbed forest soils in comparable substrates. So from the ecological point of view temporal priority should be given to forestral use.     

Antioxidant Activity and Characterization of Phenolic Compounds from Bacaba ( Oenocarpus bacaba Mart.) Fruit by HPLC-DAD-MS(n)

The phytochemicals in fruits have been shown to be major bioactive compounds with regard to health benefits. Bacaba ( Oenocarpus bacaba Mart.) is a native palm fruit from the Brazilian savannah and Amazon rainforest that plays an important role in the diet of rural communities and is also a source of income for poor people. This paper reports the characterization and analyses of phenolics from bacaba fruit extract. The total phenolic content of bacaba fruit amounted to 1759.27 ± 1.01 mg GAE/100 g, the flavonoid content was 1134.32 ± 0.03 mg CTE/100 g, and the anthocyanin content was 34.69 ± 0.00 mg cyn-3-glc/100 g. The antioxidant activity was evaluated through different assays [ORAC, FRAP, DPPH, TEAC, and cellular antioxidant assay (CAA) assays] and revealed a significant antioxidant capacity for bacaba in comparison to the data available in the literature. The assignment of the phenolic compounds using HPLC-DAD-MS(n) was based on the evaluation of their UV-vis absorption maxima (λ(max)) and mass spectral analyses, and 14 compounds were tentatively identified. The results suggest that bacaba fruits are a promising source of phenolics.     

NEPTUN,PAPA, Vergleichs- und Demonstrationsbetriebe oder Wer hat da noch den Durchblick?

The Institute for Strategies and Technology Assessment of the Julius Kühn-Institute operates four different networks: demonstration farms on IPM, reference farms for plant protection, panel pesticide applications and NEPTUN farms. All networks deal with the use of plant protection products in agricultural practice. The latter three also have very similar tasks and are closely interlinked. As a result, outsiders have significant problems distinguishing between those four farm networks and assigning the respective activities to the “right” corresponding network.Therefore, the publication aims at presenting comparatively the tasks and objectives of the various networks and to explain to the reader what are they doing, what are the differences, as well as why and how the establishment of these four different farm networks has come about.     

Erhebungen zur Anwendung von chemischen Pflanzenschutzmitteln in Zuckerrüben

Freely available information on the actual use of chemical plant protection products (PPPs) in agriculture is highly necessary for a number of scientific questions and political discussions. Therefore, since 2000, regular surveys on the use of PPPs have been carried out for the most important agricultural and horticultural crops in Germany (NEPTUN projects). In 2011, they were adjusted to legal framework changes. Since then they are known as PAPA surveys with “PAPA” being an abbreviation for Panel Pesticide Applications. For each crop a network of farms was built up. In each network, the PPP application data are collected annually, anonymized and forwarded to the Julius Kühn-Institute (JKI).All surveys and analyses based on the panel refer to the Federal Republic of Germany. The participating farms are distributed throughout Germany proportionally to the production area per crop.In sugar beet cultivation the results of PAPA surveys show that the plant protection intensity has increased slightly in recent years comparing the PAPA results with the years 2005 to 2010. There are diverse reasons for this development. Difficult-to-control weeds occurred on an increasing acreage in recent years. At the same time, an early appearance of foliar diseases (approximately beginning of July) combined with warm and humid weather during the following weeks leads to the development of the main pathogen causing leaf spot diseases in sugar beet (Cercospora beticola) in many regions. A continuing development of leaf spots increases the need for repeated fungicide applications. A high infestation with aphids was the dominating reason for increased insecticide applications in some years. Additional influences on the treatment index are due to changes in the authorization of PPPs.     

Heat stability of strawberry anthocyanins in model solutions containing natural copigments extracted from rose (Rosa damascena Mill.) petals

Thermal degradation and color changes of purified strawberry anthocyanins in model solutions were studied upon heating at 85 degrees C by HPLC-DAD analyses and CIELCh measurements, respectively. The anthocyanin half-life values increased significantly due to the addition of rose (Rosa damascena Mill.) petal extracts enriched in natural copigments. Correspondingly, the color stability increased as the total color difference values were smaller for anthocyanins upon copigment addition, especially after extended heating. Furthermore, the stabilizing effect of rose petal polyphenols was compared with that of well-known copigments such as isolated kaempferol, quercetin, and sinapic acid. The purified rose petal extract was found to be a most effective anthocyanin-stabilizing agent at a molar pigment/copigment ratio of 1:2. The results obtained demonstrate that the addition of rose petal polyphenols slows the thermal degradation of strawberry anthocyanins, thus resulting in improved color retention without affecting the gustatory quality of the product.     

Fate of sulfadiazine administered to pigs and its quantitative effect on the dynamics of bacterial resistance genes in manure and manured soil

The veterinary antibiotic sulfadiazine (SDZ), labelled by ¹⁴C, was administered to pigs to follow the fate of the drug and its metabolites in manure and manure-amended soil, and to investigate the dynamics of drug effects on resistance genes and bacterial communities. In the manure sampled over 10 days, more than 96% of the drug was found as parent compound or metabolites N-acetyl-SDZ and 4-hydroxy-SDZ. While the manure was stored the concentration of SDZ increased by 42% due to deacetylation of the metabolite N-acetyl-SDZ, whereas the minor metabolite 4-hydroxy-SDZ kept constant. In the soil the extractable amounts of the compounds decreased exponentially to less than 1 mg kg^?1 within 11 days after manure amendment. The abundances of SDZ resistance genes sul1 and sul2 were determined by qPCR relative to 16S rRNA genes in total DNA from manure and manure-amended soil. In manure both genes increased exponentially in copy number during the first 60 days of storage, suggesting preferential growth of resistant populations. However, the abundance of sul1 and sul2 decreased below 10^?5 copies per 16S rRNA gene after 175 days. With manure high amounts of sul1 and sul2 were introduced into the soil which were reduced by more than 10 times within 24 days. Thereafter, sul1 was stably maintained in soil, while sul2 further decreased between day 60 and day 165. A mathematical model was developed that could well explain the time course of sul gene abundance by considering the cost of sul genes, horizontal gene transfer, and selection of the resistant populations in the presence of SDZ. Modelling revealed a selective effect of SDZ on sul2 even at low concentrations down to 0.15 mg kg^?1 soil. Bacterial community profiles of manure and manure-amended soil were distinct, indicating that bacteria introduced with manure do not become prominent in soil. The composition of the bacterial community in manure constantly changed during storage, but mainly during the first 10 days. Profiles of soil bacterial communities revealed only a transient perturbation by manure containing SDZ.