External factors involved in the regulation of an extracellular proteinase synthesis in Bacillus megaterium

Summary A mathematical model was formulated to describe the kinetics and stoichiometry of growth and proteinase production in Bacillus megaterium. Synthesis of the extracellular proteinase in a batch culture is repressed by amino acids. The specific rate of formation of the enzyme (r _E) can be described by the formula {ie373-1}, where k ₂ and k ₃ stand for the non-repressible and repressible part of enzyme synthesis respectively, k _{S 2} is a repression coefficient and S ₂ indicates the concentration of amono acids; the values of k ₂ and k _{S 2} depend on the composition of the mixture of amino acids. Even in a high concentration, a single amino acid is less effective than a mixture of amino acids. The dependence of the proteinase repression on the concentration of an external amino acid (leucine) follows the same course as its rate of incorporation into proteins, approaching saturation at concentrations higher than 50 M (half saturation approximately 10 M). However, the total uptake of leucine did not exhibit any saturation even at 500 M external concentration.Symbols X biomass concentration, g/l - E proteinase concentration, unit/l - t time, h - S ₁ concentration of glucose, g/l - S ₂ concentration of amino acids, g/l - specific growth rate, l/h - rE specific rate of enzyme production, unit/g/h - k ₁ growth kinetic constant, l/h - k ₂ product formation kinetic constant (for non-repressible part of enzyme synthesis), unit/g - k ₃ product formation kinetic constant (for repressible portion of enzyme synthesis), unit/g - k _{S 1} saturation constant, g/l - k _{S 2} repression coefficient for a certain amino acid or amino acids mixture, g/l     

Physiological activity of immobilized cells of Claviceps fusiformis during long-term semicontinuous cultivation

Summary The 550-day semicontinuous cultivation of Claviceps fusiformis immobilized in calcium alginate is documented. The vegetative mycelium from seed or from early-production submerged culture is the best choice for immobilization. No extracellular glucans are produced by immobilized cells. Immobilized spores give low yields of clavine alkaloids. Alginate concentrations in a range of 2%–4% do not influence yield and spectrum of alkaloids. The cytoplasm of the immobilized cells becomes condensed (after 3 days), polysaccharides disappear, and centres of lipid synthesis are formed in the cytoplasm. After 60 days the cells harbour a great number of lipid particles, mitochondria are diminishing and their cristae partly disappear, indicating a decreased respiration capacity. After 350–500 days the volume of most cells is increased many times and the cells are filled with large oval bodies of electrondense material. Chloramphenicol protects immobilized cultures against bacterial contamination.     

Expression of the threonine operon fromEscherichia coli inBrevibacterium flavum andCorynebacterium glutamicum

Summary The threonine operon fromEscherichia coli was cloned in plasmid pBR322, subcloned into the shuttle vector pCEM300 and the resulting recombinant plasmid was transferred intoBrevibacterium flavum andCorynebacterium glutamicum. The expression ofE. coli threonine genes in these coryneform bacteria was demonstrated by complementing thethrA andthrB mutations and by assaying homoserine dehydrogenase activity.     

Plasmid cloning vectors replicating in Escherichia coli,amino acid-producing coryneform bacteria and Methylobacillus sp.

Summary Four hybrid plasmids were constructed from the cryptic plasmid pAM330 (from Brevibacterium lactofermentum; 4.5 kb) and the broadhost-range plasmid pGV1106 (9.0 kb; Km^r Sm^r) isolated from Escherichia coli. All of them were mobilized from E. coli into the Gram-negative methylotrophic bacterium Methylobacillus sp. and two of these constructs (pCEM300 and pCEM400) were transferred by transformation into B. flavum and Corynebacterium glutamicum. Their kanamycin-resistance determinant coming from Gram-negative hosts was expressed in these Gram-positive bacteria. Both pCEM300 and pCEM400 are very stably maintained in B. flavum and represent suitable vectors for gene cloning in coryneform producers of amino acids.     

Effect of Staphylococcus aureus serine proteinase on human lymphocyte impairment

Assessment of lymphocyte surface membrane integrity by means of trypan exclusion test and longer term survival studies based on 3HTdR uptake and Con A stimulation test were used to estimate effect of Staphylococcus aureus serine proteinase on human lymphocytes in vitro. At concentration of proteinase as high as 10 micrograms/ml, the percentage of trypan-stained lymphocytes increase with enzyme level. In contrast, decrease of 3HTdR level and Con A stimulation is already apparent at concentrations of proteinase ten times lower. Autologous serum exerts strong protective effect. Observed impairment is discussed as significant for local response in inflammation.     

Protein phosphorylation in Streptomyces albus

The phosphorylated proteins of Streptomyces albus, radioactively labeled with [32P]orthophosphate have been analyzed by gel electrophoresis and autoradiography. More than 10 protein species were found to be phosphorylated. With [32P]ATP as substrate cell free extracts phosphorylated endogenous proteins in vitro which were predominantly phosphorylated in vivo. From cell extract which exhibited active phosphorylated in vitro, a protein kinase has been partially purified. The kinase activity was identified in fractions corresponding to a 90 kDa protein.     

Immobilization of Escheria alcalescens cells with aspartate ammonia-lyase activity

Summary Immobilization of Escherichia alcalescens cells into genu-carrageenan gel for L-aspartic acid production was studied with respect to the optimized preparation of heterogenous biocatalyst /2.5–3.0% genu-carrageenan, 15% biomass, 50–55 °C, tannin added/.     

Nomenclatural remarks on the association names ofNardus stricta-rich communities in Central Europe

The present paper gives comments on the nomenclature of associations ofNardetalia andNardo-Caricion rigidae from Central Europe. A brief list of important homonyms and ambiguous names is added.     

Polypeptide cytolytic toxins from sea anemones (Actiniaria)

Abstract Biochemical and biological properties of 30 cytolytic polypeptide toxins isolated from 18 species of sea anemones ( Actiniaria ) are presented and classified into three groups according to their molecular mass, isoelectric points and the molecular mechanism of action. Phospholipase A₂-like toxins (30 kDa) from Aiptasia pallida are dissimilar to acidic metridiolysin (80 kDa) from Metridium senile and the group of about 27 predominantly basic toxins, having a molecular mass of 16–20 or 10 kDa, inhibited by sphingomyelin. They are lethal for both invertebrates and vertebrates, cardiotoxic, cutolytic and cytotoxic. Pharmacological activities, cytotoxic and cytolytic properties are mediated, at least in part, by forming pores in lipid membranes. Channels, 1–2 nm in diameter, formed in planar lipid membranes are cation selective and rectified. The mechanisms and some characteristics of ion channel formation by the toxins in the cells as well as in artificial lipid membranes are summarized and discussed in view of the structure-function studies of the toxins. Putative biological roles of toxins, based on their channel-forming activity, in the capture and killing of prey, digestion, repelling of predators and intraspecific spatial competition are suggested.