Streptomyces rimosus extracellular proteases 3. Isolation and characterization of leucine aminopeptidase

Summary A leucine aminopeptidase was purified to homogeneity fromStreptomyces rimosus culture filtrates, which are waste broth of oxytetracycline bioproduction process. Purification procedure includes ultrafiltration and chromatography on CM-Sephadex, AH-Sepharose and FPLC Mono S column. The enzyme is a monomer with molecular weight of 27,500 Daltons and pI of 7.3, stable in broad pH range and up to 70°C. It is a metallo enzyme dependent on Ca²⁺ ions for its full activity. By its specificity it is a true aminopeptidase active on amino acid amide, arylamide, peptide and ester bonds. The hydrolysing activity shows preference for leucine at the N-terminal position of substrates, also acts on aromatic acids and methionine, but does not release glycine, proline, acidic amino acids orD-amino acid residues.     

Effect of the polysaccharide ofAgrobacterium radiobacter on the growth of plants and occurrence of damping-off in sugar beet

Agrobacterium radiobacter, strain B6 (a strain isolated in this laboratory, which limited the occurrence of damping-off of sugar beet and influenced growth of plants in hot-house and field experiments) was found to produce an acidic exopolysaccharide in a mineral medium with various carbon sources. Hydrolyzates of the polysaccharide contained glucose, galactose, glycerol, succinic acid and pyruvic acid, whose quantitative content varied according to the carbon source used. The polysaccharide isolated from the medium containing glucose exhibited the highest physiological activity. Seeds germinated best and sugar beet roots were found to grow most rapidly in a medium containing 0.2 % (W/W) of the polysaccharide. The roots exposed for 3 d in this medium grew 2.7-fold as compared with non-treated plants. Higher sumbers of microorganisms were detected on the surface of roots treated with the polysaccharide. Growth of roots was also stimulated when immersing the seeds (30 min) in a 0.2 –0.4 % solution of this polysaccharide. After a two-fold treatment the roots were less damaged by the fungusPythium ultimum. Plants from seeds treated with the polysaccharide grew in the field soil more rapidly than the non-treated plants but worse than after bacterization of the seeds byA. radiobacter B6 and were only partially protected against the damping-off of sugar beet.     

Cholinesterase activity in some species of theAllium genus

In partly purified protein complexes obtained from 22 species of theAllium genus and 6 cultivars ofAllium cepa the activity of cholinesterases was detected and measured using the method of Ellman et al. The degree of its inhibition with 10^-4 M neostigmine was also tested. It was found that the activity of cholinesterase differed in individual species up to two hundred times, while the differences in the inhibitory activity of 10^-4 M neostigmine occurred only in a few cases. Individual sections and cultivars could not be characterized on the basis of the differences in the activities of the cholinesterases. Of all the sections that ofPhyllodolon shows the highest average activity. In the case of the tested cultivars distinctly the lowest activity was observed in cv. Kastická. The values of the enzymatic activity measured by Ellman’s method in this plant material include the activity of specific and unspecific cholinesterases and the part uninhibitable by neostigmine.     

Annotated chromosome counts of czechoslovak plants (31–60) (Materials for “Flóra ČSSR”—3)

Chromosome counts of the following 30 taxa (106 populations) are given:Betonica officinalis (2n=16);Bidens frondosus (2n=48);Calamagrostis arundinacea (2n=28+0–2B);Dianthus carthusianorum subsp.latifolius (2n=30);Festuca gigantea (2n=42, 42+2B);Hypericum perforatum (2n=32);Koeleria macrantha (2n=28);Kohlrauschia prolifera (2n=30);Lilium martagon (2n=24+0–2B);Melica ciliata (2n=18);Poa remota (2n=14);Ranunculus polyanthemos (2n=16);R. sardous subsp.sardous (2n=16);Roegneria canina (2n=28+0–1B);Rudbeckia laciniata (2n=76);Scabiosa canescens (2n=16);Serratula tinctoria (2n=22);Seseli elatum subsp.heterophyllum var.beckii (2n=18);S. hippomarathrum (2n=20);Thlaspicaerulescens caerulescens subsp.tatrense (2n=14);Trifolium alpestre (2n=16);T. avense (2n=14);T. medium (2n=79, 80+0–2B, 82);T. rubens (2n=16);Veronica officinalis subsp. alpestris (2n=36);Vincetoxicum hirundinaria (2n=22);Vulpia bromoides (2n=14);Zerna benekenii (2n=28)Z. monoclada (2n=28+0–8B);Z. ramosa (2n=42). Remarks on taxonomy, nomenclature and chorology for some of these taxa are given.     

Responses of cell populations of three chlorococcal algae to the action of streptomycin

Time doses of a single concentration of streptomycin and its concentration doses acting for the same time period in a liquid medium had different effects on three strains of chlorococcal algae. This concerned both the physiological responses and permanent changes in the characteristics of cell colonies growing from treated cells. Significant differences were recorded in: the number of autospores produced during the first division of the treated cells on the surface of a solid medium, the length of the lag phase, the growth rate of the diameter of cell colonies, and the survival of the treated cells. The permanent changes in the characteristics of the growing colonies were very different in the individual algal strains in quality and frequency. Physiological and the mutation effects were compared and discussed.     

Contribution to the study on the reparative effect of exogenous DNA in the irradiated meristem ofVicia faba

The present report is focused on the study of participation of exogenous DNA in the process of postirradiation reparation of meristematic cells ofVicia faba primary roots. It is aimed at comparison of the positive reparative effect of isologous DNA with postirradiation action of heterologous DNA in its native, thermally denatured and DNAase-degraded forms, or DNA degraded by ultrasound, and with the effect of other biologically important macromolecules (RNA, histone, heparin, and dextran sulphate). For this purpose, the roots ofVicia faba irradiated by 150 r exposure were cultivated in solutions containing the above substances for an appropriate time interval. In squash slides both mitotic activity of the investigated cell population and frequency of postmetaphase chromosomal aberrations induced by radiation were evaluated. It was shown that a stimulation of cell division and reparation of chromosome damages were supported exclusively by isologous DNA. On the contrary, exogenously applied heterologous DNA increased postirradiation frequency of aberrations; maintenance of native structure of applied DNA was an essential condition for the above effect. Other macromolecules investigated on the course of postirradiation reparation ofVicia faba meristematic cells were without effect.