Localization of alkaline phosphatase in bacillus intermedium Cells

Alkaline phosphatase, an enzyme secreted by Bacillus intermedius S3-19 cells to the medium, was also detected in the cell wall, membrane, and cytoplasm. The relative content of alkaline phosphatase in these cell compartments depended on the culture age and cultivation medium. The vegetative growth of B. intermedius on 0.3% lactate was characterized by increased activity of extracellular and membrane-bound phosphatases. The increase in lactate concentration to 3% did not affect the activity of membrane-bound phosphatase but led to a decrease in the activity of the extracellular enzyme. Na2UPO4 at a concentration of 0.01% diminished the activity of membrane-bound and extracellular phosphatases. CoCl2 at a concentration of 0.1 mM released membrane-bound phosphatase into the medium. By the onset of sporulation, phosphatase was predominantly localized in the medium and in the cell wall. As is evident from zymograms, the multiple molecular, forms of phosphatase varied depending on its cellular localization and growth phase

Effect of pH of medium on the dielectric properties and activity of deoxyribonucleases of Bacillus amylozyma and Serratia marcescens

It has been shown that increase in the activity of deoxyribonucleases of Bac. amylozyma, strain 9 A and Serr. marcescens strain 41 over the pH range 8·0-9·0 correlates with increase in the flexibility of the macromolecules of the enzymes. Change in the conformation of the protein macromolecules of the enzymes probably ensures the steric correspondence of the active centres of the enzymes with the molecules of the substrate, which leads to rise in their activity. © 1970

Actinomycin D Influence on Biosynthesis of Extracellular Ribonucleases by Sporulating Bacteria

The influence of actinomycin D on the synthesis of extracellular ribonucleases by "Bacillus intermedius" (binase), B.pumilus (RNAse Bp) and B.amyloliquefaciens (barnase) was studied comparatively. When added during the active synthesis of the enzymes actinomycin D stimulated the biosynthesis of binase and RNAse Bp and had no influence on the barnase biosynthesis. The response of the bacillary RNAse biosynthesis to the added actinomycin D correlated with the differences in the nucleotide sequences of the genes encoding the enzymes. The Escherichia coli SURE recombinant strains carrying the plasmids with the genes of binase, RNAse Bp and barnase under different regulatory sequences, as well as the E.coli MC4100 recombinant strains carrying the plasmids with the β-galactosidase gene under the promoters of the bacillary RNAse were isolated. However, the expression of the bacillary ribonuclease genes in the E.coli recombinant strains carrying the plasmids with the genes of the enzymes, as well as the expression of the β-galactosidase gene from the promotors of the bacillary RNAses was not stimulated by actinomycin D irrespective of the dose and addition time

Production of high-molecular-weight ribonuclease Bsn from the recombinant strain of Bacillus subtilis

Background: Ribonucleases (RNases) can be used in both basic and clinical sciences, e.g. in research on developmental processes or on antiviral and antitumor therapy. RNases have great potential as therapeutic entities. On the basis of new ribonucleases new medications can be created. Bacilli synthesize two types of secretory ribonucleases, the well-studied low-molecular-weight ribonucleases and high-molecular-weight ribonucleases. Only two RNases of the second type have so far been described: RNase Bsn from B. subtilis and binase II from B. intermedius. Materials/Methods: The activity of ribonucleases was determined from the amount of the acid-soluble products of RNA hydrolysis. The cultivation media were optimized for maximum RNase production in terms of the experimental factorial design B2 using BIOPT software. Results: Our investigation of a novel secretory ribonuclease, the Bacillus subtilis RNase Bsn expressed in the recombinant B. subtilis strain 168, showed that it is synthesized in the growth retardation phase, when inorganic phosphate is exhausted in the medium. The biosynthesis of Bsn was found to be suppressed by inorganic phosphate in the medium and activated by small amounts of the transcriptional inhibitor actinomycin D. Conclusion: Our results show that the biosynthesis of the novel secretory ribonuclease Bsn in recombinant strain Bacillus subtilis 168 is subject to negative regulation by inorganic phosphate, and is activated by small doses of actinomycin D. The stimulating effect of this antibiotic is well pronounced during the active synthesis of ribonucleases, but insignificant when ribonuclease synthesis is inhibited by Pi

Investigation of the effect of bivalent metal ions and EDTA on the activity and dielectric properties of deoxyribonuclease of Bacillus amylozyma and Serratia marcescens

1. (1) Increase and reduction in the activity of deoxyribonucleases of B. amylozyma and Serratia marcescens under the influence of the ions Mg++, Mn++ Ca++ and EDTA correlate with the increase and reduction in the asymmetry of the macromolecules of the enzymes. 2. (2) The activity of the deoxyribonucleases studied was influenced by the configuration of the molecules in solution. © 1968

Action of hexaamminecobalt on the activity of Serratia marcescens nuclease

Using CD spectroscopic and kinetic analysis, a refined mechanism of Co(NH3)6 3+ action on activity of Serratia marcescens nuclease was elucidated. The mechanism was identical with previously found mechanisms of Mg2+ and C7H5O2Hg+. Similarly to Mg2+ and C7H5O2Hg+, Co(NH3)6 3+ binding to the DNA substrate induced changes in the secondary structure which resulted in changes of the enzymatic activity of the S. marcescens nuclease. Upon binding of 0.03 Co(NH3)6 3+ per DNA phosphate, highly polymerized DNA displayed A-form characteristics. The DNA transition from B-form to A-form intermediate was followed by a decrease of the nuclease activity. The diminishing nuclease activity was consistent with diminishing values of Km and Kcat. Co(NH3)6 3+ binding to the highly polymerized DNA caused a 1.7-2.8-fold decrease in Km, and 13.3-19.9 decrease in Vmax compared with Mg-DNA complex. A vast excess of Co(NH3)63+ did not affect the activity of S. marcescens nuclease if the DNA in the assay mixture remained in its B-form conformation. Preincubation of S. marcescens nuclease with Co(NH3)6 3+ did not influence the tertiary structure of the enzyme

Effects of Bacillus intermedius ribonuclease on the properties of Saccharomyces cerevisiae

Effects of Bacillus intermedius ribonuclease on the physiological, biochemical, and consumer properties of baker's yeast Saccharomyces cerevisiae were studied. This enzyme improved the yeast raising strength and increased the cell tolerance to various adverse factors. The antistress effect of RNase correlated with an earlier start of the stationary growth phase and increased trehalose pool

Concentration dependence of the effect of Bacillus intermedius ribonuclease on the yeast Saccharomvces cerevisiae

Bacillus intermedius RNase added at a low concentration (0.001 μg/ml) stimulated yeast growth, while a high RNase concentration (1500 μg/ml) was inhibitory to yeast growth. The inhibitory effect of RNase was transient and correlated with the increase in the trehalose pool of yeast cells. The number of unbudded cells in the yeast population tended to decrease under the action of low concentrations of bacillar RNase and to increase under the action of high concentrations of this enzyme. © 2000 MAIK "Nauka/Interperiodica"

Effects of Bacillus intermedius ribonuclease on properties of Saccharomyces cerevisiae

Effects of Bacillus intermedius ribonuclease on physiological, biochemical, and consumer properties of baker's yeast Saccharomyces cerevisiae were studied. This enzyme improved the yeast raising strength and increased the cell tolerance to various adverse factors. The antistress effect of RNase correlated with an earlier start of the stationary growth phase and increased trehalose pool

Concentration dependence of the effect of bacillus intermedium ribonuclease on the yeast saccharomyces cerevisiae

-Bacillus intermedius RNase added at a low concentration (0.001 |ig/ml) stimulated yeast growth, while a high RNase concentration (1500 Jig/ml) was inhibitory to yeast growth. The inhibitory effect of RNase was transient and correlated with the increase in the trehalose pool of yeast cells. The number of unbudded cells in the yeast population tended to decrease under the action of low concentrations of bacillar RNase and to increase under the action of high concentrations of this enzyme