Aspartate carbamoyltransferase from rat liver

1. Aspartate-carbamoyltransferase activity was concentrated from rat-liver preparations. Only l-aspartate, beta-benzyl-l-aspartate and beta-erythro-hydroxy-dl-aspartate were carbamoylated enzymically. The K(m) for l-aspartate and carbamoyl phosphate have been determined by three methods: colorimetric procedure, radioactive assay with [(14)C]aspartate and an assay with [(14)C]carbamoyl phosphate. 2. The K(m) for aspartate has been determined as a function of the pH; the pK of the functional group at the active site of the enzyme, pK(e), was at pH9.0. Enzymic activity was diminished in the presence of N-ethylmaleimide, p-hydroxymercuribenzoate and the heavy metals Ag(+), Hg(2+), or Zn(2+). The inhibitions could be prevented by mercaptoethanol. These findings suggested the association of a thiol group with the enzymic activity. 3. Enzymic activity was also decreased by sodium lauryl sulphate, urea and dioxan. Competitive inhibition (with l-aspartate) was manifested by maleate, succinate, oxaloacetate, beta-erythro-hydroxy-dl-aspartate and beta-benzyl-l-aspartate. The K(i) for most of these inhibitions has been determined. 4. The properties of the liver enzyme are compared with those of Escherichia coli aspartate carbamoyltransferase and the implications of the findings are discussed.     

Some factors stimulating and inhibiting pancreatic deoxyribonuclease

Для выяснения роли ДН-азы в реакции трансформации исследовали действие различных сред, применяемых при трансформации пневмококков, на активность панкреатической ДН-азы. Эти виды среды резко повышали активность фермента. Далее, было установлено, что это повышение вызывается yeast-экстрактом, входящим в состав этих сред. С помщяю спектрального анализа и пламенного спектрофотометра в yeast-экстракте были обнаружены Mg²⁺, Ca²⁺, K⁺ и Na⁺. Изучалось действие этих ионов на активность ДН-азы и было установлено, что в присутствии Mg²⁺ (5×10^?3 m)иCa²⁺(вконцентрации 2×10^?4 m) резко повышается активность фермента, тогда как прибавление к этой системе KCl в концентрации 2×10^?1, 2×10^?2, 2×10^?3 и 2×10^?4 m оказывает угнетающее действие.—Обсуждается возможное влияние этих ионов на peaкцию трансформации.     

Plant-Derived Neurotoxic Amino Acids (β-N-Oxalylamino-l-Alanine and β-N-Methylamino-l-Alanine): Effects on Central Monoamine Neurons

In the present study the subacute effects of beta-N-oxalylamino-L-alanine (BOAA) and beta-N-methylamino-L-alanine (BMAA) on CNS monoamine neurons in rats were investigated following intracisternal injections or local intracerebral administration into substantia nigra. In vitro effects of BOAA and BMAA on high-affinity synaptosomal uptake of dopamine (DA), noradrenaline (NA), and serotonin (5-HT) were also examined. Intracisternal administration of BMAA decreased NA levels in hypothalamus, whereas no effects were seen on DA or 5-HT levels. Following intranigral injections of BOAA, NA levels tended to decrease in several regions, whereas the DA levels and the levels of DA metabolites were unaffected in all regions analyzed. Loss of tyrosine hydroxylase (TH) immunoreactivity in the intranigral injection sites and the presence of TH-immunoreactive pyknotic neurons near the borders of the injection sites were observed following both BOAA and BMAA treatments. Furthermore, substance P-immunoreactive terminals in substantia nigra pars reticulata were also found to have disappeared within the lesioned area following either BOAA or BMAA injections. Incubations with both BOAA and BMAA (10(-5) M) reduced high-affinity [3H]NA uptake in cortical synaptosomes to 69% and 41% of controls, respectively, whereas the striatal high-affinity [3H]DA uptake and the cortical high-affinity [3H]5-HT uptake were unaffected by BOAA or BMAA. The results demonstrate that both BOAA and BMAA can affect central monoamine neurons, although the potency and specificity of these substances on monoamine neurons when administered acutely into cerebral tissue or liquor cerebri seem to be low. However, the in vitro studies indicate selective effects of both compounds on NA neurons in synaptosomal preparations.     

Folylpolyglutamate Derivatives of Pisum sativum L. Determination of Polyglutamate Chain Lengths by High Performance Liquid Chromatography Following Conversion to p-aminobenzoylpolyglutamates

The folypolyglutamate derivatives of pea seedlings (Pisum sativumL. cv. Homesteader) were extracted in the presence of 2-mercaptoethanoland cleaved to p-aminobenzoylpolyglutamates by treatment withZn-HCl. Azo dyes were formed by reaction with naphthylethylenediamine and purified by polyacrylamide gel chromatography. p-Aminobenzoylpolyglutamateswere regenerated from these dyes by Zn treatment and then concentratedin vacuo. These derivatives were separated according to glutamylchain length by high performance liquid chromatography on WhatmanPartisil SAX columns. The folylpolyglutamates of 4 day old peacotyledons, pea leaves and isolated chloroplasts were mainlytetra- and pentaglutamates. These and folates of shorter chainlength were labelled when seeds and aerial shoots were incubatedwith p-aminobenzoate-[¹⁴C]. Labelling of the pentaglutamatewas reduced in seeds that were imbibed in the presence of 0.1mM methotrexate. Studies of cotyledon folylpolyglutamate synthetaseshowed that polyglutamate chain length was affected by incubationtime and the concentration of tetra-hydrofolate monoglutamatein the reaction system. ¹Present address: Department of Biology, University of Lethbridge,Alberta, Canada T1K 3M4 ²Present address: Department of Horticulture, Xiong-yue AgriculturalCollege, Xiong-yue, Liaoning Province, China (Received August 4, 1989; Accepted December 5, 1989)     

The biology of Butomus umbellatus in shallow waters with fluctuating water level

Butomus umbellatus L. is a plant species typical of littoral communities of river and stream shores. It can form continuous stands in shallow reservoirs with fluctuating water level. Their expansion is promoted by: (a) intensive vegetative reproduction of plants, (b) crowded sprouting from rhizome fragments on emerged pond bottom, (c) shallow water layer in the year following summer drainage. Expansion of B. umbellatus depends on ploidy level: two cytotypes were found in the Czech and Slovak Republics, differing in their reproductive ability. Seed production of triploids is strongly limited (they are self-incompatible within clones), while diploids can be fully fertile. Nevertheless, even in diploids, the efficiency of seed reproduction under natural conditions is low. Triploids spread by intensive vegetative reproduction, which is decisive for clonal growth of populations and their regeneration after scraping of bottom surface. During seasonal development, maximum of aboveground biomass is produced in early summer, while underground biomass increases till autumn. Growth of the plants is limited by cutting before maximum underground biomass is attained, or by duck grazing.     

Growth response of Bolboschoenus maritimus ssp. maritimus and B. maritimus ssp. compactus to different trophic conditions

Bolboschoenus maritimus (L.) Palla (=Scirpus maritimus L.) forms extensive stands in the littoral zone of small fishponds and as a weed in rice and maize fields. Within the species, two subspecies are distinguished: Bolboschoenus maritimus subsp. maritimus, B. maritimus subsp. compactus. They differ in ecology, especially in their relationships with trophic conditions and salinity of habitats. To determine growth response of these two types to different nutrient levels, we compared their seasonal development under experimental cultivation at four controlled nutrient levels. Some differences between the subspecies were found to be stable, regardless of nutrient level, namely greater amount of smaller underground tubers and more extensive rhizome system in subsp. compactus compared to less numerous larger tubers and simpler rhizome system in subsp. maritimus. In response to trophic conditions,the plants of subsp. compactus were more resistant to the conditions of the highest trophic level than those of subsp. maritimus, which were stressed. This demonstrates better adaptability and spreading ability of B. maritimus subsp. compactus at high trophic levels.     

Oxidation and amidation of salicylate by Streptomyces species

Seven streptomycete strains were tested for biotransformation of salicylate. The products were identified by nuclear magnetic resonance spectroscopy and three types of conversion were found. Streptomyces cinnamonensis and Streptomyces spectabilis formed gentisate and salicylamide concurrently. Streptomyces rimosus transformed salicylate to salicylamide. Streptomyces lividans, Streptomyces coelicolor, Streptomyces griseus and Streptomyces avermitilis produced only gentisate. Time course studies of salicylate conversion by thin-layer chromatography and high pressure liquid chromatography showed that salicylamide was accumulated in the culture broth, whereas gentisate was further metabolized.Key words: salicylate, gentisate, salicylamide, biotransformation, Streptomyces spp.