Publications by authors named "Anton Zvonarev"

9 Publications

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Changes in cell wall structure and protein set in Candida maltosa grown on hexadecane.

Folia Microbiol (Praha) 2021 Apr 27;66(2):247-253. Epub 2020 Nov 27.

G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Pushchino Center for Biological Research of the Russian Academy of Sciences, prosp. Nauki 5, Pushchino, Moscow Region, 142290, Russia.

The yeast Candida maltosa is a model organism for studying adaptive changes in the structure and function of the cell wall when consuming water-insoluble nutrient sources. The cells of C. maltosa that utilize hydrocarbons contain supramolecular structures, so-called "canals" in the cell wall. Differences in protein profiles of culture liquids and cell wall extracts of C. maltosa grown on glucose and hexadecane were analyzed. Three proteins specific of cells grown on hexadecane were revealed using mass spectrometry: glycosyl hydrolase EPD2 in the culture liquid; a protein belonging to the cytochrome C family in the 0.5 mol/L NaCl extract; and PPIA_CANAL protein known as chaperone, in the 0.1% SDS extract. The possible role of these proteins in cell wall structures responsible for adaptation to hexadecane utilization is discussed.
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http://dx.doi.org/10.1007/s12223-020-00840-2DOI Listing
April 2021

Effect of Fe on inorganic polyphosphate level in autotrophic and heterotrophic cells of Rhodospirillum rubrum.

Arch Microbiol 2019 Nov 4;201(9):1307-1312. Epub 2019 Jul 4.

G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, FRC Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences, Prospect Nauki 5, Pushchino, 142290, Russia.

Inorganic polyphosphate is involved in metal homeostasis in microorganisms. The aim of the study was to reveal differences in polyphosphate metabolism of Rhodospirillum rubrum under autotrophic and heterotrophic cultivation in the presence of Fe (2.3 mg Fe L) and without Fe (traces). Heterotrophic conditions without Fe resulted in cell lysis and low biomass yield. High polyphosphate content and low exopolyphosphatase activity were observed in the cells cultivated autotrophically in the presence of Fe. The cells grown heterotrophically in the presence of Fe contained more phosphate and low-molecular polyphosphate; on the contrary, the content of the high molecular polyphosphate decreased in parallel with the increase in exopolyphosphatase activity. The possible involvement of Pi and polyphosphate to the formation of Fe-containing inclusions is discussed.
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http://dx.doi.org/10.1007/s00203-019-01697-xDOI Listing
November 2019

The Reduced Level of Inorganic Polyphosphate Mobilizes Antioxidant and Manganese-Resistance Systems in .

Cells 2019 05 15;8(5). Epub 2019 May 15.

Skryabin Institute of Biochemistry and Physiology of Microorganisms, FRC Pushchino Center for Biological Research of the Russian Academy of Sciences, pr. Nauki 5, Pushchino 142290, Russia.

Inorganic polyphosphate (polyP) is crucial for adaptive reactions and stress response in microorganisms. A convenient model to study the role of polyP in yeast is the strain CRN/PPN1 that overexpresses polyphosphatase Ppn1 with stably decreased polyphosphate level. In this study, we combined the whole-transcriptome sequencing, fluorescence microscopy, and polyP quantification to characterize the CRN/PPN1 response to manganese and oxidative stresses. CRN/PPN1 exhibits enhanced resistance to manganese and peroxide due to its pre-adaptive state observed in normal conditions. The pre-adaptive state is characterized by up-regulated genes involved in response to an external stimulus, plasma membrane organization, and oxidation/reduction. The transcriptome-wide data allowed the identification of particular genes crucial for overcoming the manganese excess. The key gene responsible for manganese resistance is encoding a low-affinity manganese transporter: Strong down-regulation in CRN/PPN1 increases manganese resistance by reduced manganese uptake. On the contrary, , the top up-regulated gene in CRN/PPN1, is also strongly up-regulated in the manganese-adapted parent strain. Phm7 is an unannotated protein, but manganese adaptation is significantly impaired in Δ, thus suggesting its essential function in manganese or phosphate transport.
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http://dx.doi.org/10.3390/cells8050461DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6562782PMC
May 2019

Inorganic polyphosphate in methylotrophic yeasts.

Appl Microbiol Biotechnol 2018 Jun 21;102(12):5235-5244. Epub 2018 Apr 21.

FRC Biotechnology, Institute of Bioengineering, Russian Academy of Sciences, pr. Shestidesyatiletiya Oktyabrya 7-1, Moscow, 117312, Russia.

Inorganic polyphosphate (polyP) is a significant regulatory and metabolic compound in yeast cells. We compared polyP content and localization, polyphosphatase activities, and transcriptional profile of polyP-related genes in industrially important methylotrophic yeasts, Hansenula polymorpha and Pichia pastoris. The increased need for phosphate, the decrease of long-chain polyP level, the accumulation of short-chain polyP, and enhanced endopolyphosphatase activity in the crude membrane fraction were observed in methanol-grown cells compared with glucose-grown cells of both species. Transcriptome analysis revealed notable differences in the expression patterns of key genes encoding proteins related to polyP metabolism. In methanol-grown cells, the genes encoding endopolyphosphatases and phosphate transporters were upregulated. The changes in polyP metabolism are probably related to the peculiarities of bioenergetics of methanol-grown cells.
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http://dx.doi.org/10.1007/s00253-018-9008-3DOI Listing
June 2018

The biosorption of cadmium and cobalt and iron ions by yeast Cryptococcus humicola at nitrogen starvation.

Folia Microbiol (Praha) 2018 Jul 19;63(4):507-510. Epub 2018 Jan 19.

Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Prospect Nauki 5, Pushchino, Moscow Region, Russia, 142290.

Yeasts Cryptococcus humicola accumulated cadmium, cobalt, and iron (~ 50, 17, and 4% of the content in the medium, respectively) from the medium containing glucose, phosphate, and 2 mmol/L of metal salts. The effects of metal absorption on the levels of orthophosphate (Pi) and inorganic polyphosphate (polyP) varied for the metals under study. The levels of Pi and polyP increased in the case of cadmium and cobalt, respectively. In the case of iron, no changes in the levels of Pi and polyP were observed. Multiple DAPI-stained polyP inclusions were observed in the cytoplasm of cadmium-containing cells. The intensity of DAPI staining of the cell wall especially increased in case of cobalt and iron accumulation.
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http://dx.doi.org/10.1007/s12223-018-0583-6DOI Listing
July 2018

Cell wall canals formed upon growth of Candida maltosa in the presence of hexadecane are associated with polyphosphates.

FEMS Yeast Res 2017 05;17(3)

Laboratory of 3-D structures of microorganisms, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Prospect Nauki, 5 Pushchino, Moscow Region, 142290 Russian Federation.

Canals are supramolecular complexes observed in the cell wall of Candida maltosa grown in the presence of hexadecane as a sole carbon source. Such structures were not observed in glucose-grown cells. Microscopic observations of cells stained with diaminobenzidine revealed the presence of oxidative enzymes in the canals. 4΄,6΄-diamino-2-phenylindole staining revealed that a substantial part of cellular polyphosphate was present in the cell wall of cells grown on hexadecane in condition of phosphate limitation. The content and chain length of polyphosphates were higher in hexadecane-grown cells than in glucose grown ones. The treatment of cells with yeast polyphosphatase PPX1 resulted in the decrease of the canal size. These data clearly indicated that polyphosphates are constituents of canals; they might play an important role in the canal structure and functioning.
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http://dx.doi.org/10.1093/femsyr/fox026DOI Listing
May 2017

Modifications of the cell wall of yeasts grown on hexadecane and under starvation conditions.

Yeast 2016 Feb 1;33(2):55-62. Epub 2015 Dec 1.

G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Russian Federation.

Electron-microscopic examinations have demonstrated local modifications in the cell wall of the yeast Candida maltosa grown on hexadecane. In our earlier studies, these modified sites, observed in other yeasts grown on oil hydrocarbons, were conventionally called 'canals'. The biochemical and cytochemical studies of C. maltosa have revealed a correlation between the formation of 'canals' and decrease in the amount of cell wall polysaccharides, glucan and mannan. The ultrathin sections and surface replicas have shown that the 'canals' are destroyed by pronase, thus indicating that a significant proportion of their content is represented by proteins. This finding was compatible with our earlier data on the localization of oxidative enzymes in 'canals' and possible participation of the 'canals' in the primary oxidation of hydrocarbons. A completely unexpected and intriguing phenomenon has been the appearance of 'canals' in the yeast C. maltosa under starvation conditions. Unlike the yeasts grown on hexadecane, mannan almost disappears in starving cells, while the quantity of glucan first decreases and then is restored to its initial level. The role of 'canals' in starving cells is as yet unclear; it is assumed that they acquire exoenzymes involved in the utilization of products of cell lysis in the starving population. In the future, 'canals' of starving cells will be studied in connection with their possible participation in apoptosis.
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http://dx.doi.org/10.1002/yea.3140DOI Listing
February 2016

Manganese tolerance in yeasts involves polyphosphate, magnesium, and vacuolar alterations.

Folia Microbiol (Praha) 2016 Jul 8;61(4):311-7. Epub 2015 Dec 8.

Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Prospect Nauki 5, Pushchino, Moscow region, 142290, Russia.

Basidiomycetous and ascomycetous yeast species were tested for manganese tolerance. Basidiomycetous Cryptococcus humicola, Cryptococcus terricola, Cryptococcus curvatus and ascomycetous Candida maltosa, Kluyveromyces marxianus, Kuraishia capsulata, Lindnera fabianii and Sacharomyces cerevisiae were able to grow at manganese excess (2.5 mmol/L), while the growth of basidiomycetous Rhodotorula bogoriensis was completely suppressed. The lag phase duration increased and the exponential growth rate decreased at manganese excess. The increase of cell size and enlargement of vacuoles were characteristics for the cells grown at manganese excess. The alterations in inorganic polyphosphate content and cellular localization were studied. L. fabianii, K. capsulata, C. maltosa, and Cr. humicola accumulated the higher amounts of inorganic polyphosphates, while Cr. terricola and Cr. curvatus demonstrated no such accumulation. The polyphosphate content in the cell wall tested by DAPI staining increased in all species under the study; however, this effect was more pronounced in Cr. terricola and Cr. curvatus. The accumulation of Mg(2+) in the cell wall under Mn(2+) excess was observed in Cr. humicola, Cr. curvatus and Cr. terricola. The accumulation of polyphosphate and magnesium in the cell wall was supposed to be a factor of manganese tolerance in yeasts.
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http://dx.doi.org/10.1007/s12223-015-0440-9DOI Listing
July 2016

The antibiotic and membrane-damaging activities of cellobiose lipids and sophorose lipids.

J Oleo Sci 2014 ;63(7):701-7

Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences.

Antibiotic activity was compared for Cryptococcus humicola cellobiose lipids, the mixture of 2,3,4-О-triacetyl-β-D-glucopyranosyl-(1→4)-(6-О-acetyl-β-D-glucopyranosyl-(1→16)-2,16-dihydroxyhexodecanoic acid and 2,3,4-О-triacetyl-β-D-glucopyranosyl-(1→4)-(6-О-acetyl-β-D-glucopyranosyl-(1→16)-2,17,18-trihydroxyoctotodecanoic acid, and the commercial sophorose lipid mixture of a mono-acetylated acidic sophorose lipid and a di-acetylated acidic sophorose lipid, both containing the C18:1 fatty acid residue. The MIC values of cellobiose lipids were 0.005 and 0.04 mg/mL for Filobasidiella neoformans and Candida tropicalis, respectively. The MIC values of sophorose lipids were 1 and 15 mg/mL for F. neoformans and C. tropicalis, respectively. MIC values for some bacteria were in the range of 10-30 mg/mL for both glycolipid preparations. Both sophorose lipids and cellobiose lipids displayed a membrane-damaging activity against F. neoformans. The treatment with these glycolipids reduces the content of ATP in the cells of test cultures and results in their staining with ethidium bromide.
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http://dx.doi.org/10.5650/jos.ess14037DOI Listing
January 2015