Production of protoplasts from the fungi Curvularia inaequalis and Trichoderma reesei

Summary Protoplast formation in Curvularia inaequalis was achieved using non-commercial and commercial snail gut enzymes or Trichoderma harzianum enzymes. The cells were grown for enzyme treatment on cellophane sheets or in liquid cultures for varying periods of time. The production of T. harzianum enzymes is discussed. The highest protoplast yields were 2.6x10⁷ protoplasts/ml enzyme solution. Protoplasts were shown to have zero to four nuclei. Protoplast regeneration was succesfully carried out in semisolid agar.     

Synthesis of methyl α-d-glucopyranosyl-(1–2)-α-d-galactopyranosyl-(1-3)-α-d-glucopyranoside and an acyclic analogue thereof for probing the carbohydrate-binding specificity of bacteriophage ϕX 174

The title trisaccharide was synthesized using methyl 1-thioglycoside building blocks. An acyclic analogue, methyl 3-O-(α-D-glycopyranosyl-oxyethyl)-α-D-glucopyranoside, which has an ethylene bridge in place of the galactosyl residue, was also synthesized.     

Accumulation of pyrophosphate in Escherichia coli. Relationship to growth and nucleotide synthesis

Pyrophosphate (PPi) content of Escherichia coli is increased manyfold when the growth of the cells is limited by inhibition of the synthesis of nucleotides. The accumulated PPi is immediately degraded when inhibition is released and growth allowed to resume. Other tested nutritional deficiencies (starvation of carbon source, sulfate or an amino acid) fail to induce PPi accumulation.     

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)