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oxidative and assimilative enzyme activities in continuous cultures of the obligate methylotroph methylobacillus flagellatum.in methanol-limited continuous cultures of the obligate methylotrophic bacterium methylobacillus flagellatum grown at rates from 0.05 to 0.63 h-1, and also in an oxyturbidostat culture of m. flagellatum growing at the rate of 0.73 h-1, levels of methanol dehydrogenase, enzymes of formaldehyde oxidation (both linear and cyclic) and assimilation (rump cycle), a number of intermediary metabolism and tca cycle enzymes and also 'dye-linked' formaldehyde dehydrogenase were determined. it was shown tha ...19911804027
genetic mapping of the obligate methylotroph methylobacillus flagellatum: characteristics of prime plasmids and mapping of the chromosome in time-of-entry units.a pulb113 (rp4::mini-mu cts) plasmid was used to generate a library of prime plasmids carrying fragments of the methylobacillus flagellatum genome. the genes carried by these prime plasmids were identified by complementation after transfer to suitably marked escherichia coli and pseudomonas aeruginosa strains. the hybrid plasmids were used for complementation mapping with a range of e. coli, m. flagellatum, and p. aeruginosa mutants. a preliminary map of the m. flagellatum genome section with se ...19902110149
[analysis of dna-dna homologies in obligate methylotrophic bacteria].the genotypic affinity of 19 bacterial strains obligately dependent on methanol or methylamine as carbon and energy sources was studied by techniques of molecular dna hybridization. the high homology level (35-88%) between motile strain methylophilus methanolovorus v-1447d and nonmotile strain methylobacillus sp. vsb-792 as well as other motile strains (pseudomonas methanolica atcc 21704, methylomonas methanolica nrrl 5458, pseudomonas sp. w6, strain a3) indicates that all of them belong to one ...19882463459
construction of hfr-like donors of the obligate methanol-oxidizing bacterium methylobacillus flagellatum kt.the transposon-loaded plasmid pas8-121, incapable of autonomous replication in gram-negative bacteria of non-enteric group, was transferred to methylobacillus flagellatum kt wild type strain mfk1. the transconjugants arose at a frequency of 10(-7) per donor cell. the majority of the transconjugants tested exhibited the transfer of all selected chromosomal markers at rather high (10(-4)-10(-6) per donor cell) but similar frequencies. only one of the obtained donors, designated mfk 64, was capable ...19892500379
[expression of the human interferon alpha f gene in the obligate methylotroph methylobacillus flagellatum kt and pseudomonas putida].the expression of human leucocyte interferon alpha f gene in plasmid plm-ifn alpha f-273 is controlled by a hybrid tac (trp-lac) promoter. a structural gene for interferon alpha f is a component of the hybrid operon lacz'-ifn alpha f-tcr, that contains an e. coli trp-operon intercystronic region. plasmid plm ifn alpha f-273--directed interferon synthesis allows to obtain about 10(7) iu/l. this plasmid was cloned in broad-host-range vector plasmid payc31. the hybrid bi-repliconed plasmid containi ...19873119998
cloning and nucleotide sequences of the homoserine dehydrogenase genes (hom) and the threonine synthase genes (thrc) of the gram-negative obligate methylotroph methylobacillus glycogenes.we have cloned the homoserine dehydrogenase genes (hom) from the gram-negative obligate methylotrophs methylobacillus glycogenes atcc 21276 and atcc 21371 by complementation of an escherichia coli homoserine dehydrogenase-deficient mutant. the 4.15-kb dna fragment cloned from m. glycogenes atcc 21371 also complemented an e. coli threonine synthase-deficient mutant, suggesting the dna fragment contained the thrc gene in addition to the hom gene. the homoserine dehydrogenases expressed in the e. c ...19948117070
two new members of the bio b superfamily: cloning, sequencing and expression of bio b genes of methylobacillus flagellatum and corynebacterium glutamicum.cloning, characterization and expression of the bio b gene of the obligate methylotrophic bacterium, methylobacillus flagellatum, are reported. a chromosomal fragment containing bio b has been isolated by complementation of a bio b- mutant of m. flagellatum. nucleotide (nt) sequence analysis of this fragment revealed the presence of an open reading frame of 966 nt identified as bio b, which is the first gene of the m. flagellatum bio cluster. gene bio b was expressed in escherichia coli and m. f ...19968917070
[characteristics of methylobacillus flagellatum kt mutants, deficient for phosphoglucoisomerase, an enzyme of the ribulose monophosphate cycle of obligate methylotrophic bacteria].the activities of the key enzymes of ribulose monophosphate cycle for formaldehyde oxidation and assimilation were tested in crude extracts from temperature sensitive mutants of obligatemethylotroph m. flagellatum kt. two mutants deficient in phosphoglucoisomerase activity were identified during this screening. phosphoglucoisomerase of t525 pgi-1 mutant was active both at permissive (30 degrees c) and nonpermissive (42 degrees c) temperatures. complete inactivation of the enzyme at 42 degrees c ...19873119997
[temperature-sensitive mutant of the obligate methylotroph methylobacillus flagellatum kt deficient in glucose-6-phosphate dehydrogenase].the common approach is developed for isolation of mutants deficient in key enzymes of ribulose monophosphate pathway for formaldehyde oxidation and assimilation by obligate methylotrophic bacteria. the approach is based on total isolation of temperature sensitive mutants and their biochemical characterization. a number of ts- mutants of obligate methylotroph m. flagellatum kt is isolated following nitrosoguanidine induced mutagenesis. the modified screening method was developed and used for iden ...19873118203
cloning, sequence and expression in escherichia coli of the methylobacillus flagellatum reca gene.by means of interspecific complementation of an escherichia coli reca- mutation with phasmids containing a gene bank from an obligate methylotroph, methylobacillus flagellatum (mf), the reca+ gene from this bacterium was identified. when expressed in an e. coli reca- host, it can function in recombination, dna repair, and prophage induction. the nucleotide sequence of the gene has been determined. the coding region consists of 1032 bp specifying 344 amino acids. the deduced reca protein structur ...19902227454
mapping of pgi and gpd genes involved in c-1 assimilation in the obligate methylotroph methylobacillus flagellatum.homologous matings with plasmids r68.45 and pulb113, and also with hfr type donor were employed for mapping pgi and gpd genes involved in c-1 metabolism in the obligate methylotroph methylobacillus flagellatum. a preliminary map of the late chromosomal region was constructed on the basis of these experimental results. the c-1 markers were linked to methionine and leucine auxotrophy and nalidixic acid resistance markers. the phenomenon of retrotransfer, or shuttle transfer of chromosomal markers ...19902105704
[regulation of phenylalanine biosynthesis in the obligate methylotroph methylobacillus m75].regulation of phenylalanine biosynthesis has been studied in the bacterium methylobacillus m75 on the level of enzymes 3-deoxy-d-arabinoheptulose-7-phosphate-synthase, chorismatmutase, prephenatdehyrataze. the dahp-synthase is shown to be synthesized constitutively and its activity is inhibited by all aromatic aminoacids and antranilate. the synthesis of chorismatmutase and prephenatdehydratase is repressed by tyrosine, the activity of the latter enzyme, besides that, is inhibited by phenylalani ...19901979836
[characteristics of the r-plasmid pm3 (incp-9) of a broad circle of hosts].a new broad host range plasmid pm3 (incp-9) was found in a facultative methylotrophic bacteria pseudomonas putida and described. the pm3 plasmid is characterized by thermo-instability in enterobacteriaceae family of bacteria at 36 degrees c or higher temperatures. it is also unable to be inherited as an autonomous element in the obligate methylotrophic bacteria methylobacillus m75. the peculiarities of plasmid inheritance make possible to use it as a tool for genetic research, for instance, to c ...19911745275
[regulation of the activity and synthesis of 3-deoxy-d-arabinoheptuloso-7-phosphate synthase in the obligate methylotroph methylobacillus flagellatum kt].the activity of the first enzyme of aromatic path 3-deoxy-d-arabino-heptuloso-7-phosphate-synthase (dahp-synthase) is regulated by retro-inhibition and is a subject of repression. analysis of partially purified preparations of the enzyme has revealed three isoenzymes: dahp-synthase-tyr, dahp-synthase-trp and dahp-synthase-phe, each of them being regulated by a corresponding amino acid. dahp-synthase-phe is a dominant isoenzyme presenting 70% of the enzyme activity, 30% inhibition of which is pos ...19911684023
bacterial diversity and community structure in an aerated lagoon revealed by ribosomal intergenic spacer analyses and 16s ribosomal dna sequencing.we investigated the bacterial community structure in an aerated plug-flow lagoon treating pulp and paper mill effluent. for this investigation, we developed a composite method based on analyses of pcr amplicons containing the ribosomal intergenic spacer (ris) and its flanking partial 16s rrna gene. community percent similarity was determined on the basis of ris length polymorphism. a community succession was evident in the lagoon, indicated by a progressive community transition through seven sam ...200111282606
[use of the recombinant bacterial strain to control blood-sucking mosquito larvae].the recombinant strain of methylobacillus flagellatum with the cloned synthesis gene cry 4b of the toxic bac. thuringiensis var. israelensis protein proved to be effective against larvae of the anopheles stephensi, an. atroparvus, an. pulcherrimus, an. superpictus, and an. sacharovi cultured in the laboratory. the use of m. flagellatum in combination with t. pyriformis may greatly expand the scope of use of the recombinant strain to control malaria mosquito larvae. their combined use shows a 6-f ...199911221007
saccharide production from methanol by transposon 5 mutants derived from the extracellular polysaccharide-producing bacterium methylobacillus sp. strain 12s.a ch3oh-utilizing bacterium that has the ability to produce extracellular polysaccharide (eps) was isolated from a soil sample, and was identified as the obligate methylotroph methylobacillus sp. strain 12s on the basis of its 16s rdna sequence and growth-substrate specificity. the eps produced by strain 12s was purified and the sugar composition was analysed by gc-ms and hplc to reveal that the eps was a heteropolymer composed of glucosyl, galactosyl, and mannosyl residues in the molar ratio 3: ...200011030570
use of stable-isotope probing, full-cycle rrna analysis, and fluorescence in situ hybridization-microautoradiography to study a methanol-fed denitrifying microbial community.a denitrifying microbial consortium was enriched in an anoxically operated, methanol-fed sequencing batch reactor (sbr) fed with a mineral salts medium containing methanol as the sole carbon source and nitrate as the electron acceptor. the sbr was inoculated with sludge from a biological nutrient removal activated sludge plant exhibiting good denitrification. the sbr denitrification rate improved from less than 0.02 mg of no(3)(-)-n mg of mixed-liquor volatile suspended solids (mlvss)(-1) h(-1) ...200414711691
isolation and characterization of a mutant defective in the production of methanol dehydrogenase from a new restricted facultative methanol-oxidizing bacterium.a new restricted facultative methanol-oxidizing bacterium methylobacillus sp. strain sk1 (dsm 8269) was subjected to insertion mutagenesis studies with the transposon tn5 to generate mutants deficient in the production of methanol dehydrogenase (mdh). the transposon was conjugally transferred into the methylotroph by using the triparental mating procedure. the mutants induced by tn5 were selected directly from a plate containing succinate by using the allyl alcohol selection procedure. the trans ...199910794599
analysis of two formaldehyde oxidation pathways in methylobacillus flagellatus kt, a ribulose monophosphate cycle methylotroph.the roles of cyclic formaldehyde oxidation via 6-phosphogluconate dehydrogenase and linear oxidation via the tetrahydromethanopterin (h4mpt)-linked pathway were assessed in an obligate methylotroph, methylobacillus flagellatus kt, by cloning, sequencing and mutating two chromosomal regions containing genes encoding enzymes specifically involved in these pathways: 6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase and methenyl h4mpt cyclohydrolase (gnda, zwf and mch). no null mut ...200010658669
organization of threonine biosynthesis genes from the obligate methylotroph methylobacillus flagellatus.the genes encoding aspartate kinase (ask), homoserine dehydrogenase (hom), homoserine kinase (thrb) and threonine synthase (thrc) from the obligate methylotroph methylobacillus flagellatus were cloned. in maxicells hom and thrc directed synthesis of 51 and 48 kda polypeptides, respectively. the hom, thrb and thrc genes and adjacent dna areas were sequenced. of the threonine biosynthesis genes, only hom and thrc were tightly linked in the order hom-thrc. the gene for thymidylate synthase (thya) f ...199910589737
purification and characterization of chaperonins 60 and 10 from methylobacillus glycogenes.two proteins belonging to the group i chaperonin family were isolated from an obligate methanotroph, methylobacillus glycogenes. the two proteins, one a groel homologue (cpn60: m. glycogenes 60 kda chaperonin) and the other a groes homologue (cpn10: m. glycogenes 10 kda chaperonin), composed a heteropolymeric complex in the presence of atp. both proteins were purified from crude extracts of m. glycogenes by anion-exchange (deae-toyopearl) and gel-filtration (sephacryl s-400) chromatography. the ...19989764760
role of copper during carbon monoxide binding to terminal oxidases.under fully reduced conditions, reassociation kinetics of co were studied in several terminal oxidases containing copper in their binuclear center. the purified paracoccus denitrificans ba3-type quinol oxidase was found to recombine with co monophasically (tau 25-30 ms) like oxidases of the bo type from escherichia coli, the caa3 type from bacillus halodurans ftu, and the bo type from methylobacillus flagellatum kt. oxidase of the aa3 type from bovine heart recombined with co monophasically at a ...19989650593
refined genetic map of the obligate methylotroph methylobacillus flagellatum.we present a refined genetic map of the obligate methylotroph methylobacillus flagellatum. new, hfr (high-frequency-of-transfer) donors, and pulsed-field gel electrophoresis, were used to determine that m. flagellatum contains one approximately 3.1-mb circular chromosome, and no plasmids. a correlation between time-of-entry units and dna length was established. using in vivo and in vitro cloning, and sequencing, a number of new genetic markers were identified and mapped; in addition, the nature ...19989613581
regulation of ammonia assimilation in an obligate methylotroph methylobacillus flagellatum under steady-state and transient growth conditions.the obligate methylotroph methylobacillus flagellatum was grown in the presence of different ammonium concentrations and the regulation of the enzymes associated with ammonium assimilation was investigated in steady-state and transient growth regimes. as the medium changed from c-limitation to dual c/n- and finally to n-limitation, the culture passed through three definite growth phases. the nadp(+)-dependent glutamate dehydrogenase (gdh) was present under ammonium limitation of the culture grow ...19979195010
characterization of a unique mutant lyse gene, originating from corynebacterium glutamicum, encoding a product that induces l-lysine production in methylophilus methylotrophus.lyse24 is an allele of lyse encoding an l-lysine exporter of corynebacterium glutamicum. the mutant gene is able to induce l-lysine production in methylophilus methylotrophus. although lyse24 has a mutation in the middle of lyse that results in chain termination, the entire lyse locus, including the region downstream of the short open reading frame, is necessary for l-lysine production. we propose that separate polypeptides are synthesized from the lyse24 locus due to reinitiation of translation ...200617151474
transformation of methylotrophic bacteria by electroporation.an efficient system for electroporation of the methylotrophic bacteria hyphomicrobium facilis, hyphomicrobium denitrificans, methylobacillus glycogenes, methylobacterium extorquens, and methylophilus methylotrophus is described. it could be demonstrated that vectors based on the broad-host-range plasmid pbbr1 could be transferred into these strains. plasmid pbbr1kan (3.9 kb), a kanamycin-resistant derivative of pbbr1, was suitable for transformation experiments in these methylotrophic bacteria. ...19979090108
identification and characterization of the pqqdgc gene cluster involved in pyrroloquinoline quinone production in an obligate methylotroph methylobacillus flagellatum.pyrroloquinoline quinone is a prosthetic group of bacterial methanol dehydrogenases as well as some alcohol and glucose dehydrogenases. genes involved in pyrroloquinoline quinone production have previously been cloned from the representatives of the alpha and gamma subdivisions of the proteobacteria. we report identification and the sequence of the pqqdgc gene cluster in the obligate methylotroph, methylobacillus flagellatum, which belongs to the beta subdivision. the deduced products of the pqq ...19968768519
organization of the methylamine utilization (mau) genes in methylophilus methylotrophus w3a1-ns.the organization of genes involved in utilization of methylamine (mau genes) was studied in methylophilus methylotrophus w3a1. the strain used was a nonmucoid variant termed ns (nonslimy). the original mucoid strain was shown to be identical to the ns strains on the basis of chromosomal digest and hybridization patterns. an 8-kb psti fragment of the chromosome from m. methylotrophus w3a1-ns encoding the mau genes was cloned and a 6,533-bp region was sequenced. eight open reading frames were foun ...19948021188
two o-type oxidases in methylobacillus flagellatum kt.two oxidases of the o-type in membranes of the methanol-grown obligate methylotroph methylobacillus flagellatum kt were distinguished. for this purpose the kinetic analysis of the laser flash-induced optical absorbance changes of co-oxidase complexes under reducing conditions was used. the ratio of these oxidases in membranes greatly depended on the phases of bacterial growth. one of the oxidases appeared to belong to the escherichia coli o-type oxidase family being more sensitive to kcn (ki = 1 ...19947945389
isolation of individual amino acids from various microbiological sources using reversed-phase high-performance liquid chromatography.a new method for the preparative isolation of individual amino acids on a milligram scale based on reversed-phase high-performance liquid chromatography (rp-hplc) after pre-column derivatization with carbobenzoxychloride (z-cl) has been developed. the chromatographic procedure was tested by the investigation of jack bean urease hydrolysate. the method has been applied to the preparative separation of z-amino acids (from 10 up to 16) obtained from protein hydrolysates of various sources (green mi ...19957795801
effects of the amplification of the genes coding for the l-threonine biosynthetic enzymes on the l-threonine production from methanol by a gram-negative obligate methylotroph, methylobacillus glycogenes.we constructed recombinant plasmids carrying the genes coding for the l-threonine biosynthetic enzymes, the hom gene, the hom-thrc genes, and the thrb genes, of a gram-negative obligate methylotroph, methylobacillus glycogenes, and examined the effects of them on the production of l-threonine from methanol. the hom gene, which encodes the homoserine dehydrogenase, and the hom-thrc genes, containing the gene coding for threonine synthase together with the hom gene, were cloned from a wild-type st ...19947765821
cloning, sequencing, and mutation of a gene for azurin in methylobacillus flagellatum kt.the gene cluster for methylamine utilization (mau genes) has been cloned from the obligate methylotrophic bacterium methylobacillus flagellatum kt. partial sequence data showed that the organization of these genes was similar to that found in methylophilus methylotrophus w3a1-ns, including the lack of a gene for amicyanin, which had been thought to be the electron acceptor for methylamine dehydrogenase in m. flagellatum kt. however, a gene encoding azurin was discovered at the 3' end of the mau ...19957635847
[methylovorus mays--novel species of aerobic, obligatory methylotrophic bacteria associated with plants].a bacterial strain utilizing methanol as the sole source of carbon and energy was isolated from the maize phyllosphere. cells are nonpigmented gram-negative motile rods that do not form spores or prosthecae and reproduce by binary fission. the strain does not require vitamins or supplementary growth factors. it is obligately aerobic and urease-, oxidase-, and catalase-positive. the optimum growth temperature is 35-40 degrees c; the optimum ph is 7.0-7.5. the doubling time is 2 h. the bacterium i ...200011315676
overproduction of l-lysine from methanol by methylobacillus glycogenes derivatives carrying a plasmid with a mutated dapa gene.the dapa gene, encoding dihydrodipicolinate synthase (ddps) partially desensitized to inhibition by l-lysine, was cloned from an l-threonine- and l-lysine-coproducing mutant of the obligate methylotroph methylobacillus glycogenes dhl122 by complementation of the nutritional requirement of an escherichia coli dapa mutant. introduction of the dapa gene into dhl122 and al119, which is the parent of dhl122 and an l-threonine producing mutant, elevated the specific activity of ddps 20-fold and l-lysi ...200111425723
methylobacillus flagellatus kt contains a novel cbo-type cytochrome oxidase.the o-type oxidase from the methanol-grown obligate methylotroph methylobacillus flagellatus kt has been purified to homogeneity. the complex is composed of four subunits (57, 40, 35 and 30 kda). it contains six haems (4c:1b:1o) and one copper atom per molecule. it is proposed that the haem o-cu(b) binuclear centre and a low-spin haem b are located in subunit i (57 kda), two haems c reside in the cytochrome c homodimer (35 kda), two haems c belong to the dihaem cytochrome c (30 kda). the present ...200111557051
effect of growth conditions on the synthesis of terminal oxidases in methylobacillus flagellatus kt.membranes of the obligate methylotroph methylobacillus flagellatus kt contained hemes b, o, and c and cytochromes b, o, and c both in batch and in continuous cultures. neither heme a nor heme d was detected in the membranes. the cytochromes o and bb were the main components reversibly binding carbon monoxide (co) in the terminal part of the respiratory chain. the alpha-region and especially the alpha-peaks at 568 and 573 nm and the alpha-troughs at 586 and 592 on the co-difference spectra were d ...200211811956
methylobacillus arboreus sp. nov., and methylobacillus gramineus sp. nov., novel non-pigmented obligately methylotrophic bacteria associated with plants.two newly isolated obligate methanol-utilizing bacteria (strains iva(t) and lap(t)) with the ribulose monophosphate pathway of c(1) assimilation are described. the isolates are strictly aerobic, gram negative, asporogenous, motile rods multiplying by binary fission, mesophilic and neutrophilic, synthesize indole-3-acetate. the prevailing cellular fatty acids are straight-chain saturated c(16:0) and unsaturated c(16:1) acids. the major ubiquinone is q-8. the predominant phospholipids are phosphat ...201121640537
genes involved in the synthesis of the exopolysaccharide methanolan by the obligate methylotroph methylobacillus sp strain 12s.methylobacillus sp. strain 12s produces an exopolysaccharide (eps), methanolan, composed of glucose, mannose and galactose. twenty-four orfs flanking a tn5 insertion site in an eps-deficient mutant were identified, and 21 genes (epscbakldefghijmnopqrstu) were predicted to participate in methanolan synthesis on the basis of the features of the primary sequence. gene disruption analyses revealed that epsabcefgijnop and epsr are required for methanolan synthesis, whereas epskd and epsh are not esse ...200312624205
biosynthetic uniform 13c,15n-labelling of zervamicin iib. complete 13c and 15n nmr assignment.zervamicin iib is a member of the alpha-aminoisobutyric acid containing peptaibol antibiotics. a new procedure for the biosynthetic preparation of the uniformly 13c- and 15n-enriched peptaibol is described this compound was isolated from the biomass of the fungus-producer emericellopsis salmosynnemata strain 336 imi 58330 obtained upon cultivation in the totally 13c, 15n-labelled complete medium. to prepare such a medium the autolysed biomass and the exopolysaccharides of the obligate methylotro ...200314658801
[the cytochrome cbo from the obligate methylotroph methylobacillus flagellatus kt is a cytochrome-c oxidase].the oxidase cho of methylobacillus flagellatus kt was purified to homogeneity by nondenaturing gel electrophoresis, and the kinetic properties and substrate specificity of the enzyme were studied. ascorbate and ascorbate/n,n,n',n'-tetramethyl-p-phenylenediamine (tmpd) were oxidized by cbo with a ph optimum of 8.3. when tmpd served as electron donor for the oxidase cho, the optimal ph (7.0 to 7.6) was determined from the difference between respiration rates in the presence of ascorbate/tmpd and o ...200415198024
methylobacillus pratensis sp. nov., a novel non-pigmented, aerobic, obligately methylotrophic bacterium isolated from meadow grass.strain f31t was isolated from meadow grass (poa trivialis l.) sampled from the city park in helsinki. analysis of phenotypic and genotypic properties showed the strain to be related to the group of obligately methylotrophic non-methane utilizing bacteria (methylobacteria) with the ribulose monophosphate pathway of formaldehyde assimilation. phylogenetic analysis showed the strain to be closely related to the genus methylobacillus, and analysis of fatty acid composition confirmed this association ...200415388694
a top-down proteomics approach for differentiating thermal resistant strains of enterobacter sakazakii.thermal tolerance has been identified as an important factor relevant to the pathogenicity of enterobacter sakazakii in human neonates. to identify a biomarker specific for this phenotypic trait, intact protein expression profiles of 12 strains of e. sakazakii were obtained using liquid chromatography mass spectrometry. proteins were extracted from the bacterial cells, separated by reversed-phase liquid chromatography and mass analyzed. at the end of the chromatography run, the uncharged masses ...200516196092
production of aldehyde oxidases by microorganisms and their enzymatic properties.in order to establish an efficient process to decompose environmentally toxic aldehydes, dioxygen-dependent aldehyde oxidase (alod) from microorganisms was first sought, and some bacteria and actinomycetes were found to produce the enzyme in their cells. methylobacillus sp., pseudomonas sp. and streptomyces moderates were selected as the representative alod-producing strains and their enzymes were partially purified and characterized. the three alods could oxidize a wide range of aldehydes inclu ...200216233281
cloning and sequencing of the aldehyde oxidase gene from methylobacillus sp. ky4400.the aldehyde oxidase genes (aods) from methylobacillus sp. ky4400 were cloned, and sequenced. the sequences for small (aods, 489 bp), medium (aodm, 993 bp), and large (aodl, 2,328 bp) subunit genes were determined. at least one additional orf was indispensable for the expression of enzyme activity. the structural genes contained two [2fe-2s] centers, an fad binding site, and a molybdenum cofactor binding site.200516377905
molecular cloning of the dna gyrase genes from methylovorus sp. strain ss1 and the mechanism of intrinsic quinolone resistance in methylotrophic bacteria.the genes encoding the dna gyrase a (gyra) and b subunits (gyrb) of methylovorus sp. strain ss1 were cloned and sequenced. gyra and gyrb coded for proteins of 846 and 799 amino acids with calculated molecular weights of 94,328 and 88,714, respectively, and complemented escherichia coli gyra and gyrb temperature sensitive (ts) mutants. to analyze the role of type ii topoisomerases in the intrinsic quinolone resistance of methylotrophic bacteria, the sequences of the quinolone resistance-determini ...200516404155
identification of acetate- or methanol-assimilating bacteria under nitrate-reducing conditions by stable-isotope probing.stable-isotope probing (sip) was used to identify acetate- or methanol-assimilating bacteria under nitrate-reducing conditions in activated sludge. a sludge sample obtained from wastewater treatment systems was incubated in a denitrifying batch reactor fed with synthetic wastewater containing [(13)c]acetate or [(13)c]methanol as the main carbon source and nitrate as the electron acceptor. we analyzed how growth of bacterial populations was stimulated by acetate or methanol as the external carbon ...200616897304
organization of methylamine utilization genes (mau) in 'methylobacillus flagellatum ' kt and analysis of mau mutants.the organization of genes involved in utilization of methylamine (mau genes) was studied in the obligate methylotroph 'methylobacillus flagellatum' kt. nine open reading frames were identified as corresponding to the genes maufbedaglmn. in addition, an open reading frame (orf-1 encoding a polypeptide with unknown function was identified upstream of the mau gene cluster. subclones of the 'm. flagellatum' kt gene cluster were used for complementation of a series of chemically induced mau mutants o ...19979202457
expression and purification of htpx-like small heat shock integral membrane protease of an unknown organism related to methylobacillus flagellatus.the m48 conserved family of peptidases contains a single catalytic zinc ion tetrahedrally co-ordinated by two histidines within an hexxh motif. the proteases of this class are generally toxic to the cell and thus difficult to express and purify. here, we report the expression and purification of the small htpx-like heat shock metalloprotease from an unknown organism related to the obligate methylotrophic anaerobic bacterium, methylobacillus flagellatus. the protease was expressed in the escheric ...200717239953
genome of methylobacillus flagellatus, molecular basis for obligate methylotrophy, and polyphyletic origin of methylotrophy.along with methane, methanol and methylated amines represent important biogenic atmospheric constituents; thus, not only methanotrophs but also nonmethanotrophic methylotrophs play a significant role in global carbon cycling. the complete genome of a model obligate methanol and methylamine utilizer, methylobacillus flagellatus (strain kt) was sequenced. the genome is represented by a single circular chromosome of approximately 3 mbp, potentially encoding a total of 2,766 proteins. based on genom ...200717416667
characterization of an iron- and manganese-containing superoxide dismutase from methylobacillus sp. strain sk1 dsm 8269.a superoxide dismutase was purified 62-fold in seven steps to homogeneity from methylobacillus sp. strain sk1, an obligate methanol-oxidizing bacterium, with a yield of 9.6%. the final specific activity was 4,831 units per milligram protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. the molecular weight of the native enzyme was estimated to be 44,000. sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 23,10 ...200717646712
cloning and expression of methylovorus mays no. 9 gene encoding gamma-glutamylmethylamide synthetase: an enzyme usable in theanine formation by coupling with the alcoholic fermentation system of baker's yeast.gamma-glutamylmethylamide synthetase (gmas), found in an obligate methylotroph, methylovorus mays no. 9, can form theanine from glutamic acid and ethylamine in a mixture in which yeast sugar fermentation is coupled for atp regeneration. the internal and n-terminal amino acid sequences of gmas had certain similarities to putative glutamine synthetase type iii (gs iii) of methylobacillus flagellatus kt. from the m. mays no. 9 genomic dna library, a clone containing a 6.5-kbp insertional dna fragme ...200818175924
[establishment of the screening method and isolation of pqq producing strains].pyrroloquinoline quinone (pqq) is a cofactor of some oxido-reductases with many important physiological effects and potential pharmaceutical applications. the glucose dehydrogenase of escherichia coli, being a candidate for enzymic detection of pqq, is known to be a quinoprotein which is obligately dependant on pqq as cofactor. the gdh gene of e. coli was amplified and cloned into plasmid pet28a. the recombinant gdh was overexpressed in soluble form in e. coli bl21 (de3). a bioassay method was e ...200718271250
insights and inferences about integron evolution from genomic data.integrons are mechanisms that facilitate horizontal gene transfer, allowing bacteria to integrate and express foreign dna. these are important in the exchange of antibiotic resistance determinants, but can also transfer a diverse suite of genes unrelated to pathogenicity. here, we provide a systematic analysis of the distribution and diversity of integron inti genes and integron-containing bacteria.200818513439
growth of the obligate methylotroph methylobacillus flagellatum under stationary and nonstationary conditions during continuous cultivation.the growth characteristics of a chemostat culture of the obligate methylotrophic bacterium methylobacillus flagellatum have been determined. steady-state cultures growing at a rate of 0.73-0.74 h(-1), equal to the maximal growth rate, were obtained under oxyturbidostat cultivation conditions. the response of a chemostat culture to a pulse increase of methanol concentration was studied. it was shown that slow and rapidly growing cultures of m. flagellatum responded differently to pulse methanol a ...199218600999
dissecting microbial employment. 200818779810
isolation of a methylobacillus sp. that degrades microcystin toxins associated with cyanobacteria.sludge from cyanobacteria-salvaged yard in meiliang bay, lake taihu in wuxi, china was cultured and acclimated by inoculating microcystins (mcs) extract. strain j10 was isolated by degrading the mc-rr and mc-lr and was identified as methylobacillus sp. further research showed that both mc-lr and mc-rr could be completely degraded at 17h after inoculation of j10, and the degradation probably was mediated by oxygen. different enzymes, oxygen-dependent as well as oxygen-independent, with mc-degradi ...200919748604
expressed genome of methylobacillus flagellatus as defined through comprehensive proteomics and new insights into methylotrophy.in recent years, techniques have been developed and perfected for high-throughput identification of proteins and their accurate partial sequencing by shotgun nano-liquid chromatography-tandem mass spectrometry (nano-lc-ms/ms), making it feasible to assess global protein expression profiles in organisms with sequenced genomes. we implemented comprehensive proteomics to assess the expressed portion of the genome of methylobacillus flagellatus during methylotrophic growth. we detected a total of 1, ...201020639322
stoichiometric model for evaluating the metabolic capabilities of the facultative methylotroph methylobacterium extorquens am1, with application to reconstruction of c(3) and c(4) metabolism.a stoichiometric model of central metabolism was developed based on new information regarding metabolism in this bacterium to evaluate the steady-state growth capabilities of the serine cycle facultative methylotroph methylobacterium extorquens am1 during growth on methanol, succinate, and pyruvate. the model incorporates 20 reversible and 47 irreversible reactions, 65 intracellular metabolites, and experimentally-determined biomass composition. the flux space for this underdetermined system of ...200211920446
[use of methylotrophic bacteria methylobacillus flagellatum kt for isolation of deuterated exogenous carbohydrates].the cultivation conditions optimal for biosynthesis of exogenous polysaccharides by methylotrophic bacteria methylobacillus flagellatum were evaluated. the mutant strain most active in accumulating exogenous polysaccharides was selected. gradual adaptation of this strain to the deuterated medium containing 1 vol % cd3od in deuterium oxide intensified biosynthesis of exogenous polysaccharides and inhibited bacterial growth (compared to the standard medium). the monomeric composition of exogenous ...200212325295
purification and some properties of a blue copper protein from methylobacillus sp. strain sk1 dsm 8269.a blue protein was purified from the methylobacillus sp. strain sk1 that is grown on methanol in the presence of copper ion. this protein was found to be a monomer with a molecular weight of 13,500. the isoelectric point of the protein was estimated to be 8.8. the spectrum of the protein that was treated with ferricyanide showed a broad peak around 620 nm, but that of the dithionite-treated protein revealed no peaks. it contained 0.83 mol of edta-stable copper per mol protein. under air, the pro ...200212442893
genomes of three methylotrophs from a single niche uncover genetic and metabolic divergence of methylophilaceae.the genomes of three representatives of the family methylophilaceae, methylotenera mobilis jlw8, methylotenera versatilis 301 and methylovorus glucosetrophus sip3-4, all isolated from a single study site, lake washington in seattle, were completely sequenced. these were compared to each other and to the previously published genomes of methylobacillus flagellatus kt and an unclassified methylophilales strain htcc2181. comparative analysis revealed that the core genome of methylophilaceae may be a ...201121622745
an integrated proteomics/transcriptomics approach points to oxygen as the main electron sink for methanol metabolism in methylotenera mobilis.methylotenera species, unlike their close relatives in the genera of methylophilus, methylobacillus and methylovorus neither exhibit the activity of methanol dehydrogenase, nor possess mxafi genes encoding this enzyme, yet they are able to grow on methanol. in this work, we integrated two genome-wide -omics approaches, shotgun proteomics and shotgun transcriptome sequencing (rna-seq) of methylotenera mobilis jlw8 to identify genes and enzymes potentially involved in methanol oxidation, with spec ...201121764938
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