Reaction kinetics of cephalexin synthesizing enzyme from Xanthomonas citri

In enzymatic synthesis of cephalexin from D-alpha-phenylglycine methyl ester (PGM) and 7-amino-3-deacetoxy-cephalosporanic acid (7-ADCA) using alpha-acylamino-beta-lactam acylhydrolase from Xanthomonas citri, it was found that this enzyme catalyzes all three reactions including PGM hydrolysis, cephalexin synthesis, and cephalexin hydrolysis. Based on our experimental results, a mechanistic kinetic model for cephalexin synthesizing enzyme system having acyl-enzyme intermediate was proposed. From this kinetic model, the reaction rate equations for three reactions were derived, and the kinetic parameters were evaluated. A good agreement between the simulation results and the experimental results was found.     

The distribution and interconversion of ammonium and nitrate in the Skallingen salt marsh (Denmark) and their exchange with the adjacent coastal water

The potential nitrogen sources for the primary production in the intertidal area are nitrogen compounds obtained from mineralization in the sediment and the water column, nitrogen fixation, outflow from rivers and groundwater seeping from the mainland. The available inorganic nitrogen in the adjacent coastal waters decreases from 50–80 μmol NO₃ ^-/l and 6–15 μmol NH₄ ⁺/l in early spring to ca one tenth during the growing season. In the sediment of the tidal flats available ammonia and nitrate vary between 50 and 100 μmol/1 pw. In the salt marsh available ammonia increases from 200–300 nmol NH₄ ⁺/g fwt to approximately double the amount, and the available nitrate varies from 100–300 nmol NO₃ ^-/g fwt (250–750 μmol NO₃ ^-/l pw) to ca one third during the growing season. The exchange of NH₄ ⁺, NO₂ ^- and NO₃ ^- across the sediment water interface has been estimated during tidal cycles under light and dark conditions on the tidal flats. The flux of nitrogen was dependent on the flora and fauna as well as the time of the year. The tidal activity, frequency and length of inundation are considered the driving force in a two-way process between salt marshes and adjacent coastal waters. The role of marsh sediment, tidal water and sediments of the tidal flats as sites of accumulation, consumption and remineralization of organic matter is emphasized. The possible exchange of ammonia and nitrate between the salt marsh and the different compartments of the tidal water is discussed.     

Extractive bioconversion in aqueous two-phase systems

Summary The transformation of hydrocortisone to prednisolone was studied in aqueous two-phase systems, as a model for the extractive bioconversion of fine chemicals. The bacterium, Arthrobacter simplex, was able to grow in the two-phase system and the cells could be revitalized after a period of use. Use of aqueous two-phase systems made it possible to operate the reaction at higher substrate concentrations than in pure buffer solution. An adsorptive method to remove the product from the top phase was tested and shown to be both efficient and compatible with the overall process. In order to reduce the costs of operation in aqueous two-phase systems, a cheaper starch-based polymer, Reppal-PES, was successfully used as a substitute for dextran.Dedicated to Professor Dr. Georg Manecke on the occasion of his 70th birthday     

Modification of 144Ce tissue deposition by mixed ligand complex treatment in mice

The individual effect of desferrioxamine-B (DFOA), Na3Ca-diethylene-triaminepentaacetic acid (DTPA), DL-penicillamine (PA) and Na-salicylate (SA) has been examined as well as the effect of mixed-ligand treatment on the retention and elimination of 144Ce in mice. It was found that 144Ce could easily be mobilized by a single dose of DTPA. Mixed-ligand (MLCs) treatment did not change the deposition characteristics and translocation kinetics of 144Ce.     

The carbohydrate of human thrombin: Structural analysis of glycoprotein oligosaccharides by mass spectrometry

The carbohydrate structure of human thrombin has been determined by direct probe mass spectrometry of the oligosaccharides released by trifluoroacetolysis from the asialo glycoprotein. The free oligosaccharides were studied as permethylated and N-trifluoroacetylated oligosaccharide alditols. The structure was confirmed by sequential exoglycosidase digestion of intact thrombin and sugar and methylation analysis of the oligosaccharides by gas-liquid chromatography-mass spectrometry. The results indicate the following structure:

with Fuc present on only about 50% of the oligosaccharides.     

Colorless Sulfur Bacteria, Beggiatoa spp. and Thiovulum spp., in O2 and H2S Microgradients

The interactions between colorless sulfur bacteria and the chemical microgradients at the oxygen-sulfide interface were studied in Beggiatoa mats from marine sediments and in Thiovulum veils developing above the sediments. The gradients of O₂, H₂S, and pH were measured by microelectrodes at depth increments of 50 μm. An unstirred boundary layer in the water surrounding the mats and veils prevented microturbulent or convective mixing of O₂ and H₂S. The two substrates reached the bacteria only by molecular diffusion through the boundary layer. The bacteria lived as microaerophiles or anaerobes even under stirred, oxic water. Oxygen and sulfide zones overlapped by 50 μm in the bacterial layers. Both compounds had concentrations in the range of 0 to 10 μmol liter⁻¹ and residence times of 0.1 to 0.6 s in the overlapping zone. The sulfide oxidation was purely biological. Diffusion calculations showed that formation of mats on solid substrates or of veils in the water represented optimal strategies for the bacteria to achieve a stable microenvironment, a high substrate supply, and an efficient competition with chemical sulfide oxidation. The continuous gliding movement of Beggiatoa cells in mats or the flickering motion of Thiovulum cells in veils were important for the availability of both O₂ and H₂S for the individual bacteria.     

The pancreatic -cell recognition of insulin secretagogues. II. Site of action of tolbutamide

The distribution of ³⁵S-labelled tolbutamide was studied in microdissected pancreatic islets of obese-hyperglycemic mice. These islets contain more than 90 % β-cells. A comparison with the uptake of ³H-labelled sucrose, mannitol, or 3-O-methyl-D-glucose revealed that tolbutamide did not enter the β-cells but was restricted to the extracellular space. It is suggested that the β-cell plasma membrane contains a tolbutamide receptor, which is responsible for the recognition of sulfonylureas as insulin secretagogues.     

Lipid pattern and Na+−K+-dependent adenosine triphosphatase activity in the salt gland of duck before and after adaptation to hypertonic saline

Summary Ducks (Anas platyrhynchos) were fed hypertonic saline for eight days, resulting in an activation and hypertrophy of the salt gland. The Na⁺–K⁺-dependent adenosine triphosphatase, an enzyme generally assumed to be part of the active Na transport system, increased its specific activity by about 200% during this activation. Sulfatides, the major glycolipids of the salt gland, increased their concentration to the same extent. Cholesterol, cerebrosides, and six phospholipid classes showed an increase of 20–80%.A preliminary report on this work was given at the Second International Meeting of the International Society for Neurochemistry, Milan, September 1–5, 1969, and at the XIIIth International Conference on the Biochemistry of Lipids, Athens, September 7–12, 1969.