Direct assay for phosphotransacetylase and acetyl-coenzyme A carboxylase by high-performance liquid chromatography

A simple and specific assay to measure the activity of two coenzyme A derivative-processing enzymes, i.e., phosphotransacetylase (EC 2.3.1.8) and acetyl-coenzyme A carboxylase (EC 6.4.1.2), is described. The assay is based on the HPLC analysis of the short-chain coenzyme A derivatives formed by the enzymatic reaction, viz., acetyl-CoA and malonyl-CoA. For this purpose, ion-pair reversed-phase HPLC conditions are optimized. Furthermore, the influence of several variables on the enzyme reaction is studied in order to get maximum activity. Due to its short analysis time, good selectivity, and chromatogram information, HPLC proves to be an excellent method for the assay of these enzymes.     

Large-scale production of a therapeutic protein in transgenic tobacco plants: effect of subcellular targeting on quality of a recombinant dog gastric lipase

A recombinant dog gastric lipase with therapeutic potential for the treatment of exocrine pancreatic insufficiency was expressed in transgenic tobacco plants. We targeted the protein using two different signal sequences for either vacuolar retention or secretion. In both cases, an active glycosylated recombinant protein was obtained. The recombinant enzymes and the native enzyme displayed similar properties including acid resistance and acidic optimum pH. The proteolytic maturation and the specific activity of the recombinant proteins, however, were found to be dependent on subcellular compartmentalization. Expression levels of recombinant dog gastric lipase were about 5% and 7% of acid extractable plant proteins for vacuolar retention and secretion respectively. This expression system already has allowed the production of tens of grams of purified lipase through open-field culture of transgenic tobacco plants.     

A simple method for determining the sensitivity of micro-organisms to serial dilutions of antibiotics

Summary At this laboratory the routine sensitivity tests of micro-organisms against sulfathiazol and antibiotics have been performed in plastic dishes for half a year. The dishes are cleaned and sterilized in the same way as bacteriological glassware. The use of these dishes offers some advantages over other conventional dilution methods employing tubes or Petri dishes with solid media: the sensitivity to serial dilutions of one antibiotic may be measured in one and the same plate, and in this way a considerable amount of glassware and of space in the incubator may be saved; the readings are easily done and distinct. It has been demonstrated that sensitivity tests with two standard strains against sulfathiazol and various antibiotics may be reproduced with the same endpoints.     

The generalization of local staphylococcal infection in X-irradiated rabbits

Описанные опыты доказываьт, что у кроликов, облученных соответственной дозой лучей Х, возможен смертельный сепсис после заражения стафилококками в кожу—в отличие от необлученных кроликов, у которых смертельный стафилококковый сепсис удается вызвать преимущественно только интравенозной инъекцией. Сепсис развивается не сразу после инокуляции, а требует обязательно определенного латентного периода, в течение которого микроб размножается во внутренних органах животного. В принципе его патогенез не отличается от развития сепсиса у здоровых животных. Кожные некрозы, которые возникают при инокуляции стафилококков, равиваются вне зависимости от облучения.     

Effect of glucose on the decomposition of organic materials added to soil

The effect of glucose on the decomposition of plant material in soil was studied. Soil samples were enriched with different fractions of labelled alfalfa: the soluble dialyzable fraction, the soluble nondialyzable fraction, cellulose-lignin, lucerne meal. The added substances were decomposed for 42 days. One soil sample in every experimental variant was then percolated with water and another with 200 ml. 0.5% glucose for another 21 days. It was found that the effect of glucose on decomposition of the plant material depended on the latter’s composition and degree of decomposition. It was small in soils pre-enriched with soluble alfalfa fractions, decomposition of which was largely completed within the given time. Glucose markedly inhibited the release of radioactive carbon from the celluloselignin fraction and stimulated the formation of radioactive carbon dioxide from alfalfa meal.