Trypsin-/Chymotrypsin-Inhibitoren in Samen der Puffbohne (Vicia faba)

Zusammenfassung Durch isoelektrisches Fokussieren wurden mehrere Trypsin/Chymotrypsin-Inhibitoren in Samen vonVicia faba nachgewiesen, deren isoelektrische Punkte bei pH 8,5 (Hauptpeak mit Schulter) und bei pH 9,1 (Nebenpeak mit Schulter) sowie im Bereich von pH 9,1-6,4 (Nebenpeaks mit deutlich geringeren Aktivitäten) liegen. Aus einem durch Affinitätschromatographie an trägergebundenem Trypsin erhaltenen Inhibitorgemisch wurden drei Inhibitoren durch preparative Elektrophorese in Polyacrylamidgel isoliert, die sich bei pH 9,2 und bei pH 4,0 als elektrophoretisch einheitlich erwiesen. Die Molekulargewichte liegen bei 6000 Dalton. Der Anteil an basischen Aminosäuren ist hoch, während Methionin und Isoleucin fehlen. Die Hemmwirkung ist spezifisch auf Serinproteinasen gerichtet: Neben Trypsin und Chymotrypsin werden einige mikrobielle Enzyme dieser Gruppe gehemmt. Die thermische Stabilität ist relativ groß. Die Inhibitoren ausVicia unterscheiden sich damit in einer Reihe von Eigenschaften deutlich von den Inhibitoren ausPhaseolus.

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Inhibitors for trypsin and chymotrypsin in seeds of the broad bean (Vicia faba)

Summary Several inhibitors for trypsin and chymotrypsin were detected in seeds ofVicia faba by isoelectric focussing. Their isoelectric points are at pH 8.5 (main peak with shoulder), at pH 9.1 (minor peak with shoulder) and in the range of pH 9.1-6.4 (several minor peaks with significant lower activities). From the mixture of inhibitors obtained by affinity chromatography on carrier bound trypsin, three inhibitors were isolated by preparative electrophoresis in polyacrylamide gel. On electrophoresis these inhibitors behaved uniformly at pH 9.2 and at pH 4.0. Their molecular weights are about 6000 daltons. The amount of basic amino acids is high, while methionine and isoleucine are absent. Besides of trypsin and chymotrypsin some serine proteinases of microbial origin are inhibited. The thermal stability is quite high. TheVicia inhibitors therefore differ significantly from thePhaseolus inhibitors in several properties.

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Der AIF und dern Forschungskreis der Ernährungsindustrie danken wir für die finanzielle Unterstützung     

Investigation of nanoparticles by atomic and lateral force microscopy

Metallic nanoparticles have been produced on vitreous carbon substrates by means of thermal evaporation. From pictures of the particles, made by a high-resolution scanning electron microscope (HRSEM), a semispherical shape is suggested due to the total mass of deposited material. Atomic force microscopy (AFM) has been applied to this sample in order to get direct topographic information. The AFM has been operated with normal and super tips, the latter having a smaller cone angle and radius, thus following more precisely the contours of an object. Simultaneously lateral-force microscopic (LFM) images have been recorded. Major differences between the contents of HRSEM- and AFM-images are considered, emphasizing the important influence of the tips' geometry. Both the AFM and LFM line scans have been compared with and have qualitatively agreed with those calculated under simplifying assumptions.     

Multi-objective optimization of generalized reliability design problems using feature models—A concept for early design stages

Reliability optimization problems such as the redundancy allocation problem (RAP) have been of considerable interest in the past. However, due to the restrictions of the design space formulation, they may not be applicable in all practical design problems. A method with high modelling freedom for rapid design screening is desirable, especially in early design stages. This work presents a novel approach to reliability optimization. Feature modelling, a specification method originating from software engineering, is applied for the fast specification and enumeration of complex design spaces. It is shown how feature models can not only describe arbitrary RAPs but also much more complex design problems. The design screening is accomplished by a multi-objective evolutionary algorithm for probabilistic objectives. Comparing averages or medians may hide the true characteristics of this distributions. Therefore the algorithm uses solely the probability of a system dominating another to achieve the Pareto optimal set. We illustrate the approach by specifying a RAP and a more complex design space and screening them with the evolutionary algorithm.     

Functional studies of synthetic gramicidin hybrid ion channels in CHO cells

The function of a gramicidin hybrid ion channel in living Chinese hamster ovary (CHO) cells was investigated by the patch clamp method. The synthetic ion channel 1 consists of two cyclohexyl ether amino acids that link two mini-gramicidin strands. With 1 at a concentration of 1.0 microM, an increase in the whole-cell membrane conductance was observed after 1.37 min. The conductance showed larger currents when Cs(+) was used as charge carrier than when Na(+) and K(+) were used. In single-channel recordings with Cs(+) as charge carrier, the substance showed comparable single-channel amplitudes in the membrane of living cells and artificial black lipid bilayers. In addition to functioning as a cation channel, compound 1 appeared to be a water channel. Exposure of the CHO cells to an extracellular hypoosmotic solution did not substantially change the cell volume. Extracellular hypoosmotic conditions in the presence of 1 increased the cell size to 146.5 % that of the control. Thus, the synthetic hybrid channel 1 can function as a cation channel with some Cs(+) specificity, and as a water channel in CHO cells.     

Application of Immobilized α-Chymotrypsin to Peptide Synthesis in Frozen Aqueous Systems

In order to combine the high potential of frozen state peptide synthesis and the advantages of the application of immobilized proteases, the capability of carrier-bound α-chymotrypsin (CT, EC 3.4.21.1) to form peptide bonds in frozen aqueous reaction mixtures was investigated. The properties of the support materials strongly influenced the peptide yields. CT bound to rather hydrophobic supports catalysed peptide synthesis as effectively as the soluble enzyme. The immobilized enzyme preparations could be successfully re-used at least 15 times. Peptide synthesis catalysed by immobilized chymotrypsin was also studied in low water organic reaction mixtures at sub-zero temperatures. The advantages and limitations of the different approaches are discussed. © 1997 SCI.     

PPy doped PEG conducting polymer films synthesized on LiFePO4 particles

In this work, polyethyleneglycole (PEG) is introduced into polypyrrole (PPy) film coated on LiFePO₄ powder particles to promote the properties of cathode material for lithium-ion batteries. The enhancement of the electrochemical activity by the substitution of a carbon with electrochemically active polymer is investigated. Films of the PPy doped with the PEG were prepared by the chemical oxidative polymerization of pyrrole (Py) monomer. PEG has been added as an additive during polymerization process to improve mechanical and structural properties of the PPy in final PPy/PEG-LiFePO₄ cathode material. Cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and galvanostatic charge/discharge measurements were employed to characterize the electrochemical properties of PPy/PEG-LiFePO₄ material. The electrochemical performance of PPy-LiFePO₄ electrodes was greatly improved by introduction of PEG into the PPy films. Charge/discharge measurements confirmed the increase in capacity when applying PEG in PPy. The morphology and particle sizes of the prepared cathode powder material were investigated by scanning electron microscopy (SEM) and particle size analysis (PSA). Distribution of PPy and PPy/PEG films onto the LiFePO₄ particles surface was studied by time of flight secondary ion mass spectrometry (TOF-SIMS). In addition to polymeric coating layer on the surface of PPy-LiFePO₄ composite particles, some PPy unequally distributed between the particles was found. The median diameter value is 4.92 μm for PPy-LiFePO₄ sample. TOF-SIMS measurements and SEM images confirmed that thickness of polypyrrole coating on LiFePO₄ particles is about 100 nm.     

Weizen w?hrend der Reifung

Zusammenfassung In Fortführung einer früheren Arbeit über chemische Veränderungen der Weizensorte Schirokko während der Reifung [1] wurden die Gliadine und Glutenine mit Hilfe der RP-HPLC näher untersucht. Für die Gliadinfraktion konnte gezeigt werden, daß im Reifungsverlauf die-Gliadine zuerst und die-Gliadine zuletzt auftreten. Zur Entwicklung des reifen, stark differenzierten-Gliadinmusters wird ein relativ langer Zeitraum von ca. vier Wochen nach der Blüte benötigt. Die Chromatogramme der Gluteninfraktion zeigen bei den HMW-Untereinheiten nur qualitative Unterschiede, während bei den LMW-Untereinheiten eine stärkere Differenzierung im Laufe der Reifung zu beobachten ist. Der Anteil der HMW-Untereinheiten am Gesamtglutenin nimmt von ca. 15% auf ca. 30% zu und der der LMW-Untereinheiten von ca. 75% auf ca. 60% ab. Die MMW-Fraktion bleibt mit ca. 10% relativ konstant. Innerhalb der HMW-Fraktion kommt es infolge unterschiedlicher Zunahme der einzelnen Komponenten zu stärkeren Verschiebungen. Während in frühen Reifungsstadien die HMW-Untereinheiten 3 und 10 deutlich stärker vertreten sind als die Untereinheiten 5, 1 und 9 (Bezeichnung entsprechend [2]), dominiert in reifem Schirokko die Untereinheit 5, gefolgt von den Komponenten 3 und 10, während die Untereinheiten 1 und 9 in deutlich geringerer Menge vertreten sind.

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Wheat during maturation: analysis of gliadins and glutenins by RP-HPLC

Summary In continuation of a study of changes in the chemical composition of the wheat variety Schirokko during ripening [Kieffer R et al. (1988) Z Lebensm Unters Forsch 187:339], the gliadins and glutenins were investigated in more detail by RP-HPLC. The chromatograms of the gliadin fraction show very clearly that the-gliadins appear first and the-gliadins last during the ripening process. A relatively long period of about four weeks after flowering is necessary for the development of the ripe, strongly differentiated,-gliadin pattern. The chromatograms of the glutenins exhibit only qualitative changes in the high-molecular-weight subunits during maturation, while the low-molecular-weight subunits show an increasing differentiation, similar to the gliadins. The proportion of high-molecular-weight subunits increases from about 15% to 30% of whole glutenin, while the low-molecular-weight subunits decrease from about 75% to 60%. The medium-molecular-weight subunits remain relatively constant in the range of 10%. Remarkable quantitative shifts between high-molecular-weight glutenins were observed during ripening. While subunits 3 and 10 [nomenclature according to Krause I et al. (1988) Z Lebensm Unters Forsch 186:398] dominate the pattern in early stages, large amounts of subunit 5, followed by decreasing amounts of subunits 3 and 10 and, after longer tires, subunits 1 and 9 are characteristic for ripe Schirokko.

     

Comparison of the quality of sous-vide and conventionally processed carrots

 The quality of "sous-vide" and conventionally processed carrots was evaluated by sensory analysis. Furthermore, changes in colour, odour, taste and texture of the "sous-vide" products during storage over 21 days at 2  °C were determined with instrumentation. According to the panel's evaluation, the sensory quality of the "sous-vide" processed carrots remained almost constant over the storage period. However, the shear force values indicated a strengthening of the texture. The colour changed from typical orange to red-orange. A slight decrease of the flavour and aroma intensity was recorded by the panel, which was confirmed by the data gained with the Electronic nose and the AromaScan. The sous-vide processed products were always preferred and therefore obtained a higher valuation even at the end of storage. In contrast to the conventionally processed carrots, only a slight reduction in the content of sucrose, fructose and glucose was determined for the sous-vide product. Received: 3 February 1998 / Revised version: 17 April 1998