Inorganic Polyphosphate and Physiological Properties of Saccharomyces cerevisiae Yeast Overexpressing Ppn2.

The effect of the yeast endopolyphosphatase Ppn2 overproduction on the metabolism of inorganic polyphosphates in Saccharomyces cerevisiae yeast was studied. Expression of the PPN2 gene under control of the strong constitutive promoter of glyceraldehyde 3-phosphate dehydrogenase gene (PKG1) led to a significant increase in the endopolyphosphatase activity stimulated by cobalt/zinc ions. This activity was present in both soluble and membrane subcellular fractions; it was higher toward long-chain polyphosphates and could be stimulated by ADP. The content of short-chain polyphosphates in the cells of the overexpressing strain was ~2.5 times higher compared to the parent strain. The cells overexpressing Ppn2 were more resistant to peroxide and alkali. The role of short-chain polyphosphates in the adaptation to these stress factors is discussed.

[Detection of AlpA and AlpB lytic endopeptidase propeptides of Lysobacter sp. XL1 by sandwich-enzyme immunoassay based on monoclonal antibodies].

The extracellular lytic endopeptidases AlpA and AlpB of the bacterium Lysobacter sp. XL1 are highly homologous and synthesized as precursors consisting of signal peptide, propeptide and mature form. In this work, two monoclonal antibodies against propeptide endopeptidase AlpA (ProA) and eleven against propeptide endopeptidase AlpB (ProB) were obtained to study the AlpA and AlpB endopeptidases secretion. The affinity constants of the antibodies against ProA were 2.9 x 10(9) and 3.5 x 10(9) M(-1), and the affinity constants of the antibodies against ProB were from 1.5 x 10(8) to 2.2 x 10(9) M(-1). The obtained antibodies did not have cross-reactivity between themselves, as well as mature forms of the enzymes. The monoclonal antibodies based sandwich-type enzyme immunoassay has been developed for measuring the propeptide in a native form. The linear range of determination ProA was 1.5-100 ng/mL with 6% error of measurement, and for determining ProB 0.2-6.25 ng/mL with 6% error. Using the developed assay, ProA and ProB propeptides have been detected in cell lysates of Lysobacter sp. XL1 in an amount 1.18 ± 0.03 ng and 0.096 ± 0.002 ng per 1 OD540 of the bacterial culture, respectively. The immunochemical assay for detection various forms of AlpA and AlpB lytic endopeptidases can be useful when dealing with issues related to their secretion into the environment.

[Effect of the proteolytic enzymes of Bacillus licheniformis and the lysoamidase of Lysobacter sp. XL1 on Proteus vulgaris and Proteus mirabilis].

Preparations of culture liquid of three Bacullus licheniformis strains (S, 103, and 60.4) and the enzymatic preparation lysoamidase from culture liquid of Lysobacter sp. strain XL1 actively lysed preliminarily autoclaved cells of gram-negative bacteria Proteus vulgaris and P. mirabilis. Living Proteus cells treated with these enzymatic preparations were lysed during their subsequent autoclaving. Inoculation of enzyme-treated Proteus cells, taken either separately or in combination with one another and polymyxin B, into a rich medium led to cell repair and restoration of viability of culture.

[A lysoamidase bacteriolytic complex. 1. Determination of the nature of interaction of enzymes and the polysaccharide, included in the complex]. / Bakterioliticheskii kompleks lizoamidaza. 1. Opredelenie prirody vzaimodeistviia fermentov i polisakharida, vkhodiashchikh v sostav kompleksa.

The polysaccharide component of the bacteriolytic enzymatic complex "lysoamidase" (M(r) 1300 kDa) has been found to contain negatively charged ionic groups and can electrostatically interact with some enzymes within the complex. At pH 8.0 optimal for bacterial lysis, such interaction promotes the stabilization of bacteriolytic enzymes, increasing their half-inactivation temperature by 8 degrees and their stability upon storage by 40%. The polysaccharide decreases the thermal stability of phosphatase but has no stabilizing effect on the metalloprotease.

[Importance of Bdellovibrio in regulating microbial cenoses and self-purification processes in domestic sewage]. / Znachenie bdellovibrionov v reguliatsii mikrobnykh tsenozov i protsessakh samoochishcheniia bytovykh stochnykh vod.

The bacterial parasite Bdellovibrio was directly proved to be involved in the regulation of microbial cenoses and in the self-purification of domestic waste waters. The incidence of heterotrophs, Gram-negative bacteria, E. coli and Bdellovibrio was followed up in dynamics in the microecological system of waste waters for ten days. In control experiments, bdellovibrions were removed using pteridine as a vibriostatic agent. In the absence of bdellovibrions, the cell number of the studied microorganisms did not increase after reaching a stationary level. In the control, the total incidence of heterotrophs decreased 1355 times, that of Gram-negative bacteria fell down 527 times, and that of E. coli cells dropped 3419 times due to the interaction between the host bacteria and Bdellovibrio. The variations in the number of interacting cells were characteristic of a two-component parasite-host system.

[Factors affecting the participation of bacteria of the genus Bdellovibrio in the self-purification processes in the Syr Darya River]. / Faktory, vliiaiushchie na uchastie bakterii roda Bdellovibrio v protsessakh samoochishcheniia reki Syrdar'i.

The effect of season, temperature and abundance of microflora on the interrelations between bdellovibrions and host-bacteria in the syr Daryo river compared with the Oka has been studied. These factors and composition of allochthonic gram-negative bacteria in the river influence on the abundance of Bdellovibrio and extent its participation in the self-purification of basins.

[Participation of bdellovibrios in sewage self-purification processes]. / Uchastie bdellovibrionov v protsessakh samoochishcheniia stochnykh vod.

The participation of bacterial parasites belonging to the genus Bdellovibrio in the processes of sewage self-purification was studied in refineries of Pushchino. The lytic activity of Bdellovibrio resulting in a decrease of the number of heterotrophic Gram-negative bacteria and E. coli in sewage was found to depend on the temperature factor influencing the intensity of interaction between the parasite and the host bacterium. The maximal p/h (parasite/host) index was found at the water temperature of 26 degrees C and the minimal one at 19 degrees C. Sewage purified at 26 degrees C and transferred to a precipitation tank contained 800 cells of E. coli in 1 ml while that purified at 19 degrees C contained 4000 cells in 1 ml. The number of interacting organisms varied in sewage which was typical of the "parasite-host" relations. Therefore, Bdellovibrio should be involved in the self-purification of sewage from the intestinal microflora at 26 degrees C; to a lesser extent, at 23.5 degrees C; and not at all, at 19 degrees C.

Antibacterial activity of seawater and microvibrion-predators (Microvibrio marinus Roscoffensis) in it.

In the seawater of the Atlantic Ocean and surface waters of different points of the World Ocean, including arctic basins and equatorial regions, microvibrion-predators of bacterial cells have been discovered. The number of microvibrions in the surface waters of the English Channel reaches a thousand cells per ml, but it varies, depending on the season. In this article a method of the separation and culture of microvibrions is described. The ultrastructure of two strains of microvibrions was investigated and a similarity found between their structure and Gram-negative bacteria. The distinctive features of microvibrions are their sea derivation and the ability to reproduce only in the presence of bacteria, without penetrating bacterial cells. To a certain extent, the antibacterial activity of seawater is connected with the presence in it of microvibrions.

[Study of marine microvibrios of Roscoff (Microvibrio marinus roscoffensis)]. / Etude sur les microvibrions marins de roscoff (Microvibrio marinus roscoffensis).

Marine microvibrios, predators of E. coli, have been detected worldwide, from equatorial waters to the polar regions. The morphology and behavior of two microvibrios have been studied. Their multiplication and aggressiveness in regard to their bacterial prevy are not impeded by dialysis membranes and manifest themselves, away from all direct contact between the predator and its prey.

[Purity criteria for Bdellovibrio bacteriovorus cultures]. / Kriterii chistoty kul'tur Bdellovibrio bacteriovorus

Two-component cultures of Bdellovibrio bacteriovorus, a bacterial parasite, are not always pure; sometimes they contain microbial forms, different from the host and parasite, which cannot be isolated by conventional techniques of inoculation on solid growth media. The only way to isolate them is to apply techniques used for the reversion of L-forms of bacteria. The isolated microorganisms have been identified. The criteria of purity were established for two-component cultures consisting of the host and parasite.