Reverse prenyltransferase in the biosynthesis of fumigaclavine C in Aspergillus fumigatus: gene expression, purification, and characterization of fumigaclavine C synthase FGAPT1

A putative prenyltransferase gene-fgaPT1-has been identified in the biosynthetic gene cluster of fumigaclavines in Aspergillus fumigatus AF293. The gene was cloned and overexpressed in Escherichia coli, and the His6-fusion FgaPT1 was purified to near homogeneity and characterized biochemically. The enzyme was found to convert fumigaclavine A into fumigaclavine C by attaching a dimethylallyl moiety to C-2 of the indole nucleus in a "reverse" manner, that is, by connection of C-3 of the dimethylallyl moiety to an aromatic nucleus. FgaPT1 is a soluble, dimeric protein with a subunit size of 50 kDa. K m(app) values for fumigaclavine A and dimethylallyl diphosphate were determined to be 6 and 13 microM, respectively, while the turnover number was 0.8 s(-1). Metal ions such as Mg2+ and Ca2+ are not essential for the enzymatic activity. FgaPT1 showed relatively strict substrate specificity towards fumigaclavine A, with only dimethylallyl diphosphate being accepted as a donor under our conditions. FgaPT1 is the first reverse prenyltransferase from fungi to have been purified and characterized in homogenous form after heterologous overproduction. Surprisingly, it shows very low sequence similarity to the recently identified prenyltransferase LtxC from cyanobacteria, which also catalyzes the reverse prenylation of an indole nucleus.     

Hydrophilic and hydrophobic materials and their applications

Wettability of a material’s surface plays a significant role in how fluids interact with such surfaces. Wetting behavior is universal but can vary depending on the chemical nature of the solid and liquid phases. Plants and animals adapt to their environment by having evolved special properties. These properties are such as hydrophilic and hydrophobic. Hydrophilic surface has a strong affinity to water and spreading of water on such surface is preferred. The degree of hydrophilicity of the substance can be measured by measuring the contact angle between the liquid and solid phases. Hydrophobic materials are known as non-polar materials with a low affinity to water, which makes them water repelling. A contact angle of less than 90° indicates hydrophilic interaction where as an angle greater than 90° indicates a hydrophobic interaction. More recently, superwetting such as superhydrophilicity has been receiving an increased focus in the literature due to its potential significance. Superhydrophilic surface has a contact angle of less than 5°.

The fabrication of hydrophilic materials can be carried out in two main ways: depositing molecules on surfaces or modification of surface chemistry. Both methods have been successful historically in achieving their intended purposes. Hydrophobic and superhydrophobic materials can be produced with many fabrication methods such as layer-by-layer assembly, laser process, the solution-immersion method, sol-gen techniques, chemical etching, and Hummer’s method.

The applications of such an important property are significant. For example, hydrophilic surfaces can be used in anti-fogging applications, biomedical, filtration, heat pipes, and many others. Hydrophobic and superhydrophobic materials have been successfully applied in many sectors, such as: (I) the removal of petroleum from aqueous solutions, (II) applied to plastic, ceramics, and mesh to contribute to the oil removal from aqueous solutions, (III) hydrophobic layers have a strong self-cleaning effect on plastics, heat pipes, metals, textiles, glass, paints, and electronics, (IV) hydrophobic layers improve the anti-freezing behavior of heat pipes which prevents unwanted build-up and (V) they function as a water and dust protecting coat on electronics.

The presence of this property is historic but there is still a huge potential for development for its applications in many sectors such as water treatment, heat transfer applications, biomedical devices, and many more.     

Bioaccessibility of provitamin A carotenoids in bananas (Musa spp.) and derived dishes in African countries

Bananas and plantains (Musa spp.) constitute an important component of the diet in Africa. Substantial levels of provitamin A carotenoids (pVACs) in Musa fruit have been reported, but the bioaccessibility of these pVACs remains unknown. In this study, we used an in vitro digestion model to assess the bioaccessibility (i.e. the transfer into micelles) of pVACs from boiled bananas and derived dishes using the Eastern Democratic Republic of Congo as a study context. In particular, the effect of different food ingredients added to boiled bananas on pVAC’s bioaccessibility was studied. The bioaccessibility of all-trans β-carotene ranged from 10% to 32%, depending on the food recipes, and was modified, particularly when pVACs-rich ingredients (palm oil/amaranth) were added. Efficiency of micellarization of all-trans β-carotene was similar to that of all-trans α-carotene and depended on the cultivar (Musilongo, plantain type, 16%; Vulambya, East African cooking type, 28%), while that of the 13-cis isomer was higher (21–33.5%). Taking into account bioaccessibility, the estimated vitamin A activity was significantly different across the different Musa-based dishes tested. Results are discussed in terms of recommendations to help reduce vitamin A deficiency in Musa-dependent African communities.     

Glucons?ureverg?rung durch Milchs?urebakterien des Weines

Zusammenfassung Insgesamt 116 verschiedene, überwiegend aus Wein isolierte Stämme von Milchsäure-bakterien der GattungenLactobacillus, Leuconostoc, Pediococcus undStreptococcus wurden auf die Fähigkeit zur Verwertung von Gluconsäure untersucht. Bis auf einzelne Ausnahmen wurde Gluconsäure von denLactobacillus-Arten, die den UntergattungenStreptobacterium undBetabacterium angehörten, und der Mehrzahl derLeuconostoc-Stämme vergoren. VonPediococcus wurde Gluconsäure nicht abgebaut. Bei wachsenden Kulturen und ruhenden Zellen vonLactobacillus brevis, Lactobacillus casei undLeuconostoc oenos waren die Endprodukte des Gluconsäureabbaus Milchsäure, Essigsäure, Äthanol und CO₂. Milchsäure entsteht aus Gluconsäure in annähernd äquimolaren Mengen. In einem aus Wein und Hefeextrakt bestehenden Medium (pH 3,5) wurden 0,5 g Gluconsäure und 4,5 g Äpfelsäure je 1 vonLactobacillus brevis undLeuconostoc oenos vollständig abgebaut.

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Fermentation of gluconic acid by lactic acid bacteria isolated from wine

Summary The fermentation of gluconic acid was investigated in 116 different strains of lactic acid bacteria belonging to the generaLactobacillus, Leuconostoc, Pediococcus andStreptococcus. Most of the strains were isolated from wine. Gluconic acid was fermented by nearly all species of the genusLactobacillus (belonging to the subgeneraStreptobacterium andBetabacterium and by most strains ofLeuconostoc. The pediococci did not ferment gluconic acid. Growing cultures and resting cells ofLactobacillus brevis, Lactobacillus casei andLeuconostoc oenos formed lactic acid, acetic acid, ethanol and CO₂ from gluconic acid. Lactic acid was formed in about equimolar amounts from gluconic acid.Lactobacillus brevis andLeuconostoc oenos metabolised completely 0.5 g of gluconic acid and 4.5 g of malic acid per liter when cultured in a medium consisting of wine and a small amount of yeast extract (at pH 3.5).

     

Metal-ion-induced formation and stabilization of protein cages based on the cowpea chlorotic mottle virus

The cowpea chlorotic mottle virus (CCMV) is a versatile building block for the construction of nanoreactors and functional materials. Upon RNA removal, the capsid can be reversibly assembled and disassembed by adjusting the pH. At pH 5.0 the capsid is in the native assembled conformation, while at pH 7.5 it disassembles into 90 capsid protein dimers. This special property enables the encapsulation of various molecules, such as protein and enzymes, but only at low pH. It is possible to stabilize the capsid at pH 7.5 by addition of negatively charged polyelectrolytes or negatively charged particles, but these methods all fill the interior of the capsid, leaving little or no space for other cargo molecules. This pH restriction therefore severely limits the range of enzymes that can be encapsulated, and hampers the investigation of the CCMV capsid as a nanoreactor for the study of enzymes in confined spaces. Herein, the interaction of N-terminal histidine-tag-modified capsid proteins with several metal ions is reported. Depending on the conditions used, nanometer-sized protein particles or capsidlike architectures are formed that are stable at pH 7.5. This metal-mediated stabilization methodology is employed to form stable capsids containing multiple proteins at pH 7.5, thereby greatly expanding the scope of the CCMV capsid as a nanoreactor.     

L?slichkeit und gel-elektrophoretische Muster hitzedenaturierter Proteine, auch von st?rkehaltigen Proben, nach Einwirkung von Na-dodecylsulfat (SDS)

Zusammenfassung Das Lösungsvermögen des Anionendetergents Na-dodecylsulfat (SDS) in Gegenwart disulfidbrückenspaltender Reagentien auf native und auf hitzedenaturierte Proteine wird an Fleisch-und Kartoffelproteinen mit Hilfe elektrophoretischer Methoden untersucht. Die zur Resolubilisierung notwendige Menge an SDS und die erforderliche Incubationszeit werden für die verschiedenen Proben ermittelt. Die Proteine des Skeletmuskels von Rind, Schwein, Huhn und Pute einerseits und die Proteine der europäischen Kartoffelsorten und der südamerikanischen Primitivkultivare andererseits zeigen in der SDS-PAGE ein sehr ähnliches Bandenmuster, das speziesdifferenzierend kaum ausgewertet werden kann, aber ein gutes Charakteristikum für Muskel-bzw. Kartoffelproteine ist.

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Solubility and electrophoretic patterns of heat-denaturated proteins (including samples containing starch) after treatment with SDS (sodiumdodecylsulfat)

Summary The ability of the anionic detergent SDS (sodium-dodecylsulfate) to solubilize native and heat-denaturated proteins of meat and potatoes in presence of disulphide reducing agents was checked by electrophoretic methods. Skeletal muscle proteins of beef, pork, chicken and turkey have shown very similar patterns in SDS-gelelectrophoresis, which could hardly be used for species identification but they are characteristic for fresh or boiled muscles. Proteins of European potato cultivars and primitive cultivars from South America have almost identical SDS-patterns even if taken from boiled potatoes. Their patterns are different from the proteins of other European crops plants and may serve for detection of the presence of potatoes in food mixtures.

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Diese Arbeit ist Teil der Dissertation von I. de Wreede. Die Analyse der tierischen Proteine bearbeitete I. de Wreede.Sonderdruckanfragen an: I. de Wreede (Adresse siehe oben)     

Enhanced mechanical properties of epoxy composites using cellulose micro- and nano-crystals

Epoxy polymers are commonly utilized in structural applications due to their high bearing capacity and excellent chemical resistance. However, their inherent brittleness poses a significant challenge for their use in high shock and fracture strength products. To address this shortcoming, fillers can be incorporated into the polymer during preparation. In this study, we aimed to investigate the effect of incorporating cellulose-based fillers, namely cellulose nanocrystals (CNCs) and microcrystalline cellulose (MCC), on the mechanical properties of epoxy polymer composites. The study evaluated the impact of various factors, including filler concentration, particle size, and moisture content, on the mechanical properties of the composites. The results demonstrated that the incorporation of CNC or MCC powders at concentrations below 5% could enhance the mechanical properties of the resulting epoxy composites without adversely affecting their surface and thermal properties. The maximum tensile strength and fracture toughness of the filler-based epoxy composites were achieved at 2 and 4 wt% for CNCs and MCC, respectively. CNCs with a smaller particle size distribution were found to be much more effective than MCC in improving the mechanical properties of the epoxy composites. Furthermore, utilizing dried fillers resulted in a higher improvement in tensile strength, which was achieved at lower filler concentrations.     

Desalination of a brick by application of an electric DC field

Salts in masonry can cause various problems as decay of the masonry itself, lost adhesion of plaster and hygroscopic moisture. Chlorides are among the most common building salts and the present paper is focused on removal of chlorides from a brick in an applied electric field as a step towards developing an electrochemical desalination method for brick masonry. Experiments were conducted in laboratory scale with one type of bricks that were contaminated with either NaCl or KCl through submersion in salt solutions prior to application of current. It was seen that NaCl was slower supplied to the brick during submersion and slower removed in the applied electric field than KCl. This indicates that the removal rate of chloride depends on the associated cation and this must be taken into account when desiding the duration of full scale actions. The electrochemical desalination was very efficient and 99% removal of chloride was obtained. The final concentration in the brick after treatment was less than 10 mg Cl/kg and this concentration is unproblematic. When low salt concentrations were reached during the electrochemical treatment, electroosmotic dewatering of the brick started, showing that electroosmotic dewatering occurs at low ionic concentrations.