The Experts below are selected from a list of 795 Experts worldwide ranked by ideXlab platform

Yuri A. Trotsenko - One of the best experts on this subject based on the ideXlab platform.

  • Ancylobacter sonchi sp. nov., a novel Methylotrophic Bacterium frоm roots of Sonchus arvensis L.
    International journal of systematic and evolutionary microbiology, 2017
    Co-Authors: N. V. Agafonova, E. N. Kaparullina, Yuri A. Trotsenko, Nina V. Doronina
    Abstract:

    An aerobic facultatively Methylotrophic Bacterium was isolated from roots of Sonchus arvensis L. and designated strain OsotT The cells of this strain were Gram-stain-negative, asporogenous, motile short rods multiplying by binary fisson. They utilized methanol, methylamines and a variety of polycarbon compounds as the carbon and energy sources. Methanol was assimilated after sequential oxidation to formaldehyde and CO2 via the ribulose bisphosphate pathway. The organism grew optimally at 22–29 °C and pH 7.5–8.0. The dominant phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol and diphosphatidylglycerol (cardiolipin). The major cellular fatty acids of strain OsotT cells grown in R2A medium were C18 : 1 ω7c (49.0 %), C19 : 0 ω8c cyclo (38.3 %) and C16 : 0 (8.4 %). The major ubiquinone was Q-10. The DNA G+C content of strain OsotT was 66.1 mol% (T m). On the basis of 16S rRNA gene sequence analysis strain OsotT is phylogenetically related to the members of genus Ancylobacter (97.1–98.8 % sequence similarity). Based on 16S rRNA gene sequence analysis and DNA–DNA relatedness (27–29 %) with type strains of the genus Ancylobacter , the novel isolate is classified as a new species of this genus and named Ancylobacter sonchi sp. nov.; the type strain is OsotT (=VKM B-3145T=JCM 32039T).

  • methylobacillus methanolivorans sp nov a novel non pigmented obligately Methylotrophic Bacterium
    International Journal of Systematic and Evolutionary Microbiology, 2017
    Co-Authors: E. N. Kaparullina, Yuri A. Trotsenko, N V Doronina
    Abstract:

    Three strains of obligately Methylotrophic Betaproteobacteria (ZT, SP and M3) with the ribulose monophosphate pathway of C1 assimilation are described. The isolates were strictly aerobic, Gram-stain-negative, asporogenous, motile (strains ZT and M3) or non-motile (strain SP) rods that multiplied by binary fisson, and were mesophilic and neutrophilic. All three strains utilized methanol but only strains SP and M3 utilized methylamine as carbon and energy sources. The prevailing cellular fatty acids were straight-chain saturated C16 : 0 and unsaturated C16 : 1 ω7c acids. The major ubiquinone was Q-8. The predominant phospholipids were phosphatidylethanolamine, phosphatidylglycerol and cardiolipin. Ammonia was assimilated by glutamate dehydrogenase. The DNA G+C contents of strains ZT, SP and M3 were 51.0, 52.0 and 52.0 mol% (T m), respectively. Levels of 16S rRNA gene sequence similarity between the three strains were very high (99.9–100 %), and they shared high levels of DNA–DNA relatedness (88–98 %). Based on 16S rRNA gene sequence analysis and DNA–DNA relatedness (19–30 %) with the type strains of the genus Methylobacillus , the novel isolates ZT, SP and M3 are classified as representing a novel species of this genus, for which the name Methylobacillus methanolivorans sp. nov. is proposed. The type strain is ZT (=VKM B-3037T=JCM 31401T=CCUG 68999T).

  • methylopila musalis sp nov an aerobic facultatively Methylotrophic Bacterium isolated from banana fruit
    International Journal of Systematic and Evolutionary Microbiology, 2013
    Co-Authors: N V Doronina, E. N. Kaparullina, Tatjana V Bykova, Yuri A. Trotsenko
    Abstract:

    A newly isolated, facultatively Methylotrophic Bacterium (strain MUSAT) was investigated. The isolate was strictly aerobic, Gram-stain-negative, asporogenous, motile, rod-shaped and multiplied by binary fission. The strain utilized methanol, methylamine and an apparently narrow range of multi-carbon compounds, but not methane, dichloromethane or CO2/H2, as the carbon and energy sources. Growth occurred at pH 5.5–9.5 (optimum, pH 7.0) and 16–40 °C (optimum, 28–30 °C). The major fatty acids of methanol-grown cells were C18 : 1ω7c, C18 : 0 and 11-methyl-C18 : 1ω7c . The predominant phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol and phosphatidylmonomethylethanolamine. The major ubiquinone was Q-10. The strain had methanol and methylamine dehydrogenases as well as the enzymes of the N-methylglutamate pathway (lyases of γ-glutamylmethylamide and N-methylglutamate). C1 assimilation occurs via the isocitrate lyase-negative serine pathway. Ammonium was assimilated by glutamate dehydrogenase and the glutamate cycle (glutamate synthase/glutamine synthetase). The DNA G+C content of the strain was 64.5 mol% (determined from the melting temperature). Based on 16S rRNA gene sequence similarity (97.0–98.9 %) and DNA–DNA relatedness (36–38 %) with representatives of the genus Methylopila ( Methylopila capsulata IM1T and Methylopila jiangsuensis JZL-4T) the isolate was classified as a novel species of the genus Methylopila , for which the name Methylopila musalis sp. nov. is proposed. The type strain is MUSAT ( = VKM B-2646T = DSM 24986T = CCUG 61696T).

  • methylopila jiangsuensis sp nov an aerobic facultatively Methylotrophic Bacterium
    International Journal of Systematic and Evolutionary Microbiology, 2011
    Co-Authors: Jinwei Zheng, N V Doronina, Baojian Hang, Yuri A. Trotsenko
    Abstract:

    The taxonomic status was determined of an aerobic, facultatively Methylotrophic strain, JZL-4(T), isolated from activated sludge. The cells were gram-negative, asporogenous, colourless, motile, short rods. The strain utilized methanol, methylamine, formate and a variety of polycarbon compounds, but not methane, dichloromethane or CO(2)/H(2), as carbon and energy sources. C(1) compounds were assimilated via the isocitrate lyase-negative serine pathway. Optimal growth occurred at 30 °C, pH 6.5-7.5 and 0.5 % (w/v) NaCl. The major cellular fatty acids were C(18 : 1)ω7c and C(18 : 0). The major phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol and phosphatidylmonomethylethanolamine (PME); PME, the main phospholipid of strain JZL-4(T), was absent or present in only minor amounts in Methylopila capsulata IM1(T), Methylopila helvetica DM9(T) and Albibacter methylovorans DM10(T). The major ubiquinone was Q-10. The DNA G+C content of strain JZL-4(T) was 70.4 mol% (T(m)). Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strain showed high sequence similarities to M. capsulata IM1(T) (97.2 %), A. methylovorans DM10(T) (94.9 %) and M. helvetica DM9(T) (94.1 %), and showed less than 94 % similarity to strains of other species with validly published names. Strain JZL-4(T) had a low level of DNA-DNA relatedness (34 %) with M. capsulata IM1(T). On the basis of phenotypic, genetic and phylogenetic data, strain JZL-4(T) is proposed to represent a novel species of the genus Methylopila, with the name Methylopila jiangsuensis sp. nov. The type strain is strain JZL-4(T) ( = ACCC 05406(T) = DSM 22718(T) = VKM B-2555(T)).

  • hansschlegelia plantiphila gen nov sp nov a new aerobic restricted facultative Methylotrophic Bacterium associated with plants
    Systematic and Applied Microbiology, 2007
    Co-Authors: Ekaterina Ivanova, N V Doronina, Yuri A. Trotsenko
    Abstract:

    A new genus, Hansschlegelia, and a new species, Hansschlegelia plantiphila, are proposed for three strains of methanol-utilizing bacteria isolated from lilac buds (strain S(1)(T)), linden buds (strain S(2)) and blue spruce needles (strain S(4)), which were selected in winter at -17 degrees C. These bacteria are aerobic, Gram-negative, colorless, non-motile short rods that multiply by binary fission and employ the ribulose bisphosphate (RuBP) and the serine pathways for C(1) assimilation. The strains have a limited number of growth substrates and use methanol, methylamine, formate, CO(2)/H(2) and glycerol as carbon and energy sources. Only strain S(1)(T) grows with ethanol and inulin. The strains are neutrophilic and mesophilic, and synthesize phytohormones (auxins and cytokinins) and vitamin B(12). Their major cellular fatty acids are saturated C(16:0), straight-chain, unsaturated C(18:1)(omega)(7) and cyclopropane C(19 cyc) acids. The main ubiquinone is ubiquinone-10 (Q-10). The dominant phospholipids are phosphatidylethanolamine, phosphatidylcholine and diphosphatidylglycerol (cardiolipin). The DNA G+C content is 68.5+/-0.2 mol%. The strains share almost identical 16S rRNA gene sequences, a high DNA-DNA hybridization value (72-86%) and represent a novel lineage of autotrophic methanol-utilizing bacteria within the Alphaproteobacteria. Collectively, these strains comprise a new genus and species H. plantiphila gen. nov., sp. nov., with strain S(1)(T) (VKM B-2347(T), NCIMB 14035(T)) as the type strain.

N V Doronina - One of the best experts on this subject based on the ideXlab platform.

  • hansschlegelia quercus sp nov a novel Methylotrophic Bacterium isolated from oak buds
    International Journal of Systematic and Evolutionary Microbiology, 2020
    Co-Authors: N. V. Agafonova, E. N. Kaparullina, Denis S Grouzdev, N V Doronina
    Abstract:

    Novel aerobic, restricted facultatively Methylotrophic bacteria were isolated from buds of English oak (Quercus robur L.; strain DubT) and northern red oak (Quercus rubra L.; strain KrD). The isolates were Gram-negative, asporogenous, motile short rods that multiplied by binary fisson. They utilized methanol, methylamine and a few polycarbon compounds as carbon and energy sources. Optimal growth occurred at 25 °C and pH 7.5. The dominant phospholipids were phosphatidylethanolamine, phosphatidylcholine, diphosphatidylglycerol and phoshatidylglycerol. The major cellular fatty acids of cells were C18 : 1 ω7c, 11-methyl C18 : 1 ω7c and C16 : 0. The major ubiquinone was Q-10. Analysis of 16S rRNA gene sequences showed that the strains were closely related to the members of the genus Hansschlegelia : Hansschlegelia zhihuaiae S113T(97.5–98.0 %), Hansschlegelia plantiphila S1T (97.4–97.6 %) and Hansschlegelia beijingensis PG04T(97.0–97.2 %). The 16S rRNA gene sequence similarity between strains DubT and KrD was 99.7 %, and the DNA–DNA hybridization (DDH) result between the strains was 85 %. The ANI and the DDH values between strain DubT and H. zhihuaiae S113T were 80.1 and 21.5 %, respectively. Genome sequencing of the strain DubT revealed a genome size of 3.57 Mbp and a G+C content of 67.0 mol%. Based on the results of the phenotypic, chemotaxonomic and genotypic analyses, it is proposed that the isolates be assigned to the genus Hansschlegelia as Hansschlegelia quercus sp. nov. with the type strain DubT (=VKM B-3284T=CCUG 73648T=JCM 33463T).

  • Methylophilus aquaticus sp. nov., a New Aerobic Methylotrophic Bacterium Isolated from a Freshwater Reservoir
    Microbiology, 2018
    Co-Authors: E. N. Kaparullina, N. V. Agafonova, Yu. A. Trotsenko, N V Doronina
    Abstract:

    — A new restricted facultative methylotroph, strain LTK^T, isolated from the water of a small freshwater reservoir, was found to carry out the KDPG variant of the ribulose monophosphate pathway of С_1 meta-bolism. The cells of the isolate were aerobic, gram-negative, non-spore-forming, nonmotile rods reproducing by binary fission. The isolate was mesophilic and neutrophilic; it synthesized indole derivatives. Hexadecanoic (C_16:0) and cis -9-hexadecenoic (C_16:1ω7) acids were the major fatty acids, and Q_8 was the major ubiquinone. Phosphatidylethanolamine and phosphatidylglycerol were the dominant phospholipids, while diphosphatidylglycerol was not present. Activities of α-ketoglutarate dehydrogenase and glutamate dehydrogenase were not revealed. Ammonium was assimilated via the enzyme system of the glutamate cycle. The 16S rRNA gene sequence of strain LTK^T exhibited high similarity to those of Methylophilus species: 99.6% to M. leisingeri DM11^T and 99.4% to M. flavus Ship^T. DNA‒DNA homology between strain LTK^T and M. leisingeri DM11^T and M. flavus Ship^T was 47 and 40%, respectively. Based on these data, strain LTK^T was assigned to the new species, Methylophilus aquaticus sp. nov. (= VKM B-3159^T = JCM 32032^T = CCUG 70602^T).

  • methylobacillus methanolivorans sp nov a novel non pigmented obligately Methylotrophic Bacterium
    International Journal of Systematic and Evolutionary Microbiology, 2017
    Co-Authors: E. N. Kaparullina, Yuri A. Trotsenko, N V Doronina
    Abstract:

    Three strains of obligately Methylotrophic Betaproteobacteria (ZT, SP and M3) with the ribulose monophosphate pathway of C1 assimilation are described. The isolates were strictly aerobic, Gram-stain-negative, asporogenous, motile (strains ZT and M3) or non-motile (strain SP) rods that multiplied by binary fisson, and were mesophilic and neutrophilic. All three strains utilized methanol but only strains SP and M3 utilized methylamine as carbon and energy sources. The prevailing cellular fatty acids were straight-chain saturated C16 : 0 and unsaturated C16 : 1 ω7c acids. The major ubiquinone was Q-8. The predominant phospholipids were phosphatidylethanolamine, phosphatidylglycerol and cardiolipin. Ammonia was assimilated by glutamate dehydrogenase. The DNA G+C contents of strains ZT, SP and M3 were 51.0, 52.0 and 52.0 mol% (T m), respectively. Levels of 16S rRNA gene sequence similarity between the three strains were very high (99.9–100 %), and they shared high levels of DNA–DNA relatedness (88–98 %). Based on 16S rRNA gene sequence analysis and DNA–DNA relatedness (19–30 %) with the type strains of the genus Methylobacillus , the novel isolates ZT, SP and M3 are classified as representing a novel species of this genus, for which the name Methylobacillus methanolivorans sp. nov. is proposed. The type strain is ZT (=VKM B-3037T=JCM 31401T=CCUG 68999T).

  • methylopila musalis sp nov an aerobic facultatively Methylotrophic Bacterium isolated from banana fruit
    International Journal of Systematic and Evolutionary Microbiology, 2013
    Co-Authors: N V Doronina, E. N. Kaparullina, Tatjana V Bykova, Yuri A. Trotsenko
    Abstract:

    A newly isolated, facultatively Methylotrophic Bacterium (strain MUSAT) was investigated. The isolate was strictly aerobic, Gram-stain-negative, asporogenous, motile, rod-shaped and multiplied by binary fission. The strain utilized methanol, methylamine and an apparently narrow range of multi-carbon compounds, but not methane, dichloromethane or CO2/H2, as the carbon and energy sources. Growth occurred at pH 5.5–9.5 (optimum, pH 7.0) and 16–40 °C (optimum, 28–30 °C). The major fatty acids of methanol-grown cells were C18 : 1ω7c, C18 : 0 and 11-methyl-C18 : 1ω7c . The predominant phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol and phosphatidylmonomethylethanolamine. The major ubiquinone was Q-10. The strain had methanol and methylamine dehydrogenases as well as the enzymes of the N-methylglutamate pathway (lyases of γ-glutamylmethylamide and N-methylglutamate). C1 assimilation occurs via the isocitrate lyase-negative serine pathway. Ammonium was assimilated by glutamate dehydrogenase and the glutamate cycle (glutamate synthase/glutamine synthetase). The DNA G+C content of the strain was 64.5 mol% (determined from the melting temperature). Based on 16S rRNA gene sequence similarity (97.0–98.9 %) and DNA–DNA relatedness (36–38 %) with representatives of the genus Methylopila ( Methylopila capsulata IM1T and Methylopila jiangsuensis JZL-4T) the isolate was classified as a novel species of the genus Methylopila , for which the name Methylopila musalis sp. nov. is proposed. The type strain is MUSAT ( = VKM B-2646T = DSM 24986T = CCUG 61696T).

  • methylopila jiangsuensis sp nov an aerobic facultatively Methylotrophic Bacterium
    International Journal of Systematic and Evolutionary Microbiology, 2011
    Co-Authors: Jinwei Zheng, N V Doronina, Baojian Hang, Yuri A. Trotsenko
    Abstract:

    The taxonomic status was determined of an aerobic, facultatively Methylotrophic strain, JZL-4(T), isolated from activated sludge. The cells were gram-negative, asporogenous, colourless, motile, short rods. The strain utilized methanol, methylamine, formate and a variety of polycarbon compounds, but not methane, dichloromethane or CO(2)/H(2), as carbon and energy sources. C(1) compounds were assimilated via the isocitrate lyase-negative serine pathway. Optimal growth occurred at 30 °C, pH 6.5-7.5 and 0.5 % (w/v) NaCl. The major cellular fatty acids were C(18 : 1)ω7c and C(18 : 0). The major phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol and phosphatidylmonomethylethanolamine (PME); PME, the main phospholipid of strain JZL-4(T), was absent or present in only minor amounts in Methylopila capsulata IM1(T), Methylopila helvetica DM9(T) and Albibacter methylovorans DM10(T). The major ubiquinone was Q-10. The DNA G+C content of strain JZL-4(T) was 70.4 mol% (T(m)). Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strain showed high sequence similarities to M. capsulata IM1(T) (97.2 %), A. methylovorans DM10(T) (94.9 %) and M. helvetica DM9(T) (94.1 %), and showed less than 94 % similarity to strains of other species with validly published names. Strain JZL-4(T) had a low level of DNA-DNA relatedness (34 %) with M. capsulata IM1(T). On the basis of phenotypic, genetic and phylogenetic data, strain JZL-4(T) is proposed to represent a novel species of the genus Methylopila, with the name Methylopila jiangsuensis sp. nov. The type strain is strain JZL-4(T) ( = ACCC 05406(T) = DSM 22718(T) = VKM B-2555(T)).

I Ignatov - One of the best experts on this subject based on the ideXlab platform.

Young Min Kim - One of the best experts on this subject based on the ideXlab platform.

  • cloning and molecular characterization of groesl heat shock operon in Methylotrophic Bacterium methylovorus sp strain ss1 dsm 11726
    Journal of Biochemistry and Molecular Biology, 2005
    Co-Authors: Chi Yong Eom, Si Wouk Kim, Eungbin Kim, Young Min Kim
    Abstract:

    Abstract The groESL bicistronic operon of a restricted facultative Methylotrophic Bacterium Methylovorus sp. strain SS1 DSM 11726 was cloned and characterized. It was found to consist of two ORFs encoding proteins with molecular masses of 11,395 and 57,396 daltons, which showed a high degree of homology to other bacterial GroES and GroEL proteins. The genes were clustered in the transcription order groES-groEL. Northern blot analyses suggested that the groESL operon is transcribed as a bicistronic 2.2-kb mRNA, the steady-state level of which was markedly increased by temperature elevation. Primer extension analysis demonstrated one potential transcription start site preceding the groESL operon, which is located 100 bp upstream of the groES start codon. The transcription start site was preceded by a putative promoter region highly homologous to the consensus sequences of Escherichia coli sigma 32-type heat shock promoter, which functioned under both normal and heat shock conditions in E. coli. Heat shock mRNA was maximally produced by Methylovorus sp. strain SS1 approximately 10 min after increasing the temperature from 30 to 42 degrees C. The groESL operon was also induced by hydrogen peroxide or salt shock.

  • cloning molecular characterization and transcriptional analysis of dnak operon in a Methylotrophic Bacterium methylovorus sp strain ss1 dsm 11726
    Molecules and Cells, 2002
    Co-Authors: Chi Yong Eom, Eungbin Kim, Sang Tae Park, Si Wook Kim, Young Min Kim
    Abstract:

    Three structural genes that consist of a dnaK operon in a restricted facultative Methylotrophic Bacterium Methylovorus sp. strain SS1 DSM 11726 were cloned and characterized. The genes were clustered in the transcription order grpE-dnaK-dnaJ. The cloned grpE, dnaK, and dnaJ genes had open-reading frames of 474, 1,926, and 1,116 nucleotides, coding for proteins with calculated molecular masses of 17,390, 69,761, and 41,050, respectively. The overall identities in the deduced amino acid sequences of GrpE, DnaK, and DnaJ with those of the Escherichia coli homologs were 45.2, 74.5, and 61.2%, respectively. Northern blot analyses with grpE-, dnaK-, and dnaJ-specific probes revealed that the three genes are co-transcribed as a 4.0-kb mRNA. A primer extension analysis revealed that the transcription of the dnaK operon started at the nucleotide A that is located 28 bp upstream of the grpE start codon. The transcription start site was preceded by a putative promoter region 15'-CCCCGCTTGAA(13-bp)CCCCAATTT-3'], which is highly homologous to the consensus sequences of the E. coli sigma32-type heat shock promoter. The putative promoter worked under both normal and heat shock conditions in E. coli. The nature of the nucleotide sequence in the second half of the -35 region played a critical role during transcription. The heat shock mRNA was maximally produced at about 10 min after transfer of the Methylovorus sp. strain SS1 from 30 to 42 degrees C. The dnaK operon was also induced by ethanol, hydrogen peroxide, and NaCl shocks. The cloned dnaK operon complemented the E. coli dnaK mutant.

O V Mosin - One of the best experts on this subject based on the ideXlab platform.

  • the biosynthesis of deuterium labeled amino acids using a strain of facultative Methylotrophic Bacterium вreviBacterium methylicum 5662 with rump cycle of carbon assimilation
    European Journal of Molecular Biotechnology, 2015
    Co-Authors: O V Mosin, D A Skladnev, I Ignatov, Vitaly Shvets
    Abstract:

    We used Gram-positive aerobic facultative Methylotrophic Bacterium, BreviBacterium methylicum, L-phenylalanine producer with ribulose-5-monophosphate (RuMP) cycle for carbon assimilation for microbiological preparation of [ 2 H]phenylalanine via conversion of low molecular weight substrates ([U- 2 H]MeOH and 2 H2O). For this purpose, the cells of the methylotroph with improved growth characteristics were used on minimal salt media M9 supplemented with 2 % (v/v) [U- 2 H]MeOH and increasing gradient of 2 Н2O concentration from 0; 24,5; 49,0; 73,5 up to 98 % (v/v) 2 Н2O. L-phenylalanine was isolated from the growth medium after adding 5 M 2 HCl (in 2 Н2О), pH = 2,0 by extraction with isopropanol and subsequent crystallization in ethanol (output 0,65 g/l). Alanine, valine, and leucine/isoleucine were produced and accumulated exogenously in amounts of 5–6 mol in addition to the main product of biosynthesis. The method allows to obtain [ 2 Н]amino acids with different levels of deuterium enrichment, depending on 2 Н2O concentration

  • deuterated Methylotrophic biomass as a substrate for microbiological synthesis of 2 h labeled purine ribonucleoside inosine by chemoheterotrophic Bacterium bacillus subtilis b 3157
    2015
    Co-Authors: O V Mosin, I Ignatov, Moscow State
    Abstract:

    We propose to use the hydrolyzed deuterated biomass of the facultative Methylotrophic Bacterium BreviBacterium methylicum B-5662 as a source of 2 H-labeled growth substrates for microbiological synthesis of 2 H-labeled purine ribonucleoside inosine, excreted into the liquid microbial culture (LC) by a Gram-positive chemoheterotrophic Bacterium Bacillus subtilis B-3157. The Bacterium was grown on heavy water (HW) medium with 2% (v/v) hydrolysate of deuterated biomass of the Methylotrophic Bacterium BreviBacterium methylicum B-5662 obtained on minimal salt media M9 supplemented with 2% (v/v) [ 2 H]methanol and increasing gradient of 2 Н2O concentration from 0; 24,5; 49,0; 73,5 up to 98% (v/v) 2 Н2O. Isolation of 2 H-labeled inosine from LC was performed by adsorption/desorption on activated carbon with following extraction by 0,3 M ammonium–formate buffer (pH = 8,9), crystallization in 80% (v/v) EtOH, and ion exchange chromatography (IEC) on a column with AG50WX 4 cation exchange resin equilibrated with 0,3 M ammonium–formate buffer and 0,045 M NH4Cl. The investigation of deuterium incorporation into the inosine molecule by FAB method demonstrated incorporation of 5 deuterium atoms into the molecule (the total level of deuterium enrichment – 65,5 atom% 2 H) with 3 deuterium atoms being included into the ribose and 2 deuterium atoms – into the hypoxanthine residue of the molecule.

  • Methylotrophic biomass as substrate for cultivation of chemoheterotrophic Bacterium bacillus subtilis b 3157
    Journal of Medicine Physiology and Biophysics, 2015
    Co-Authors: O V Mosin, I Ignatov
    Abstract:

    We propose to use a hydrolyzed deuterated biomass of the facultative Methylotrophic Bacterium BreviBacterium methylicum B-5662 as a source of 2 H-labeled growth substrates for microbiological preparation of 2 H-labeled purine ribonucleoside inosine, excreted into the liquid microbial culture (LC) by a Gram-positive chemoheterotrophic Bacterium Bacillus subtilis B-3157 . The Bacterium was grown on heavy water (HW) medium with 2% (v/v) hydrolysate of deuterated biomass of the Methylotrophic Bacterium BreviBacterium methylicum B-5662 obtained on minimal salt media M9 supplemented with 2% (v/v) [U- 2 H]MeOH and increasing gradient of 2 Н 2 O concentration from 0; 24.5; 49.0; 73.5 up to 98% (v/v) 2 Н 2 O. Isolation of 2 H-labeled inosine from LC was performed by adsorption/desorption on activated carbon with following extraction by 0.3 M ammonium–formate buffer (pH = 8.9), crystallization in 80% (v/v) EtOH, and ion exchange chromatography (IEC) on a column with AG50WX 4 cation exchange resin equilibrated with 0.3 M ammonium–formate buffer and 0.045 M NH 4 Cl. The investigation of deuterium incorporation into the inosine molecule by FAB method demonstrated incorporation of 5 deuterium atoms into the molecule (the total level of deuterium enrichment – 65.5 atom% 2 H) with 3 deuterium atoms being included into the ribose and 2 deuterium atoms – into the hypoxanthine residue of the molecule. Key words : 2 H-labeled inosine, biosynthesis, biosynthetic pathways, heavy water, Bacillus subtilis, BreviBacterium methylicum

  • using of facultative Methylotrophic Bacterium breviBacterium methylicum в 5652with rmp cycle of carbon assimilation for microbiological synthesis of 2h phenylalanine with different levels of deuterium enrichment
    European Journal of Molecular Biotechnology, 2014
    Co-Authors: I Ignatov, O V Mosin, D A Skladnev, Vitaly Shvets
    Abstract:

    With using of a strain of of L-phenylalanine secreted Gram-positive aerobic facultative Methylotrophic bacteria BreviBacterium methylicum В-5652, assimilating methanol via ribulose-5-monophosphate (RMP) cycle of carbon assimilation it was carried out the preparative microbiological synthesis of phenylalanine and metabolically related amino acids (alanine, valine, leucine/isoleucine in the amount of 5–6 mmol/l), labelled with deuterium (2H). The data on adaptation of L-phenylalanine secreted Methylotrophic Bacterium В. methylicum to the maximal concentration of deuterium in minimal growth medium M9 with 98 % 2Н2O and 2 % [2Н]methanol, and data on biosynthesis of deuterium labelled L-phenylalanine with different levels of enrichment are submitted. The developed method for biosynthesis allows to obtain [2Н]phenylalanine with different levels of isotopic enrichment, depending on 2Н2O concentration in growth media M9, from 17 % (2 deuterium atoms) (on growth medium with 24,5 % 2Н2О) right up to 75 % (6 deuterium atoms) (on growth medium with 98 % 2Н2О) with introduction of deuterium to benzyl С6Н5СН2-fragment of molecule that is confirmed with the data of the electron impact (EI) mass-spectrometry analysis of methyl ethers of N-dimethylamino(naphthalene)-5- 25 European Journal of Molecular Biotechnology, 2014, Vol.(3), № 1 sulfochloride (dansyl) derivatives of [2Н]phenylalanine, isolated from the liquid culture after its separation by RP HPLC

  • preparation of highly deuterated phenylalanine alanine valine and leucine isoleucine using facultative Methylotrophic Bacterium reviBacterium methylicum
    Journal of Medicine Physiology and Biophysics, 2014
    Co-Authors: O V Mosin, I Ignatov
    Abstract:

    Microbiological preparation of [ 2 H]amino acids was performed by conversion of low molecular weight substrates ([U- 2 H]MeOH and 2 H 2 O) by Gram-positive aerobic facultative Methylotrophic Bacterium, BreviBacterium methylicum , being L-phenylalanine producer having ribulose-5-monophosphate (RuMP) cycle for carbon assimilation. For this purpose, the cells of the methylotroph with improved growth characteristics were used on minimal salt media M9 supplemented with 2% (v/v) [U- 2 H]MeOH and increasing gradient of 2 ? 2 O concentration from 0; 24.5; 49.0; 73.5 up to 98% (v/v) 2 ? 2 O. L-phenylalanine was isolated from the growth medium after adding 5 M 2 HCl (in 2 ? 2 ?), pH = 2.0 by extraction with isopropanol and subsequent crystallization in ethanol (output 0.65 g/l). Alanine, valine, and leucine/isoleucine were produced and accumulated exogenously in amounts of 5–6 mmol in addition to the main product of biosynthesis. The method allows to obtain [ 2 ?]amino acids with different levels of deuterium enrichment, depending on 2 ? 2 O concentration in growth media, from 17% 2 ? (2 deuterium atoms) (on the growth medium with 24.5% (v/v) 2 ? 2 ?) up to 75% 2 ? (6 deuterium atoms) (on the growth medium with 98% (v/v) 2 ? 2 ?) with introduction of deuterium to benzyl ? 6 ? 5 ?? 2 -fragment of molecule that is confirmed with the data of electron impact (EI) mass spectrometry analysis of methyl ethers of N-5-dimethylamino(naphthalene)-1-sulfochloride [ 2 H]amino acids after the separation by reverse-phase HPLC. Keywords: BreviBacterium methylicum , [U- 2 H]MeOH, heavy water, biosynthesis, [ 2 H]amino acids, EI mass spectrometry, HPLC.