comparative study on conformational stability and subunit interactions of two bacterial asparaginases.the denaturation and reconstitution of erwinia carotovora and escherichia coli l-asparaginases has been followed by optical rotatory dispersion, circular dichroism and analytical ultracentrifugation. denaturation in urea results in dissociation of the native enzyme (mol. wt. 140 000 approx.) to produce unfolded subunits (mol. wt. 35 000 approx.); the erwinia l-asparaginase subunits can be refolded by dilution or dialysis in alkaline conditions, ph 10.5, without aggregation to the active tetramer ...1975237538
toxic and immunodepressive effects of l-asparaginase from e. coli and from erwinia carotovora following chronic administration in rats.the effects of the administration of l-asparaginase from e. coli and erwinia carotovora were studied in rats treated for 90 days with 800 or 3200 iu/kg body weight. the studies included overall toxicity on the liver, pancreas, and enteric mucosa as evaluated by both opitcal and electron microscopic examination, biochemical findings, behaviour of igm-hemolysin producing cells, and antias-paraginase antibody production. the toxic effect and the immunodepressive activity appeared rather early, tend ...1977341899
improvement in the therapeutic, immunological, and clearance properties of escherichia coli and erwinia carotovora l-asparaginases by attachment of poly-dl-alanyl peptides.the chemical modification of both escherichia coli and erwinia carotovora asparaginases by a dl-alanine-n-carboxyanhydride polymerization technique produced modified enzymes which had greater protease stability, retained most of their catalytic activity, and demonstrated a 7- to 10-fold prolongation in plasma clearance properties in normal mice and rats. concomitantly, plasma substrate depletion was also extended 5 to 13 days longer for the modified as compared with the native enzymes. for prepa ...1979376113
immunochemical comparison of phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase among the enterobacteriaceae.the bifunctional enzyme of the tryptophan operon, phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase (prai-ingps;ec, was characterized by an immunochemical study of six representative members of the enterobacteriaceae: escherichia coli, salmonella typhimurium, enterobacter aerogenes, serratia marcescens, erwinia carotovora, and proteus vulgaris. prai-ingps was purified from e. coli, and antisera were prepared in rabbits. these antisera were utilized in quantitativ ...1977403178
inducer of pectic acid lyase in erwinia carotovora. 1977413045
amino acid degrading enzymes for cancer therapy.the development of microbial enzymes for cancer therapy presents difficulties not commonly experienced with biological drugs. the development of the enzyme asparaginase from escherichia coli in the usa and of the serologically different asparaginase from the plant pathogen erwinia carotovora in this establishment, has not only added to the choice of antileukaemia drugs but also provided a valuable guide to the selection and development of new therapeutic enzymes. our own programme has led to the ...1977415922
comparative experimental evaluation of immunodepressive and toxic effects of l-asparaginase (nsc-109229) from escherichia coli and from erwinia carotovora. 1976769978
bacterial stalk rot of maize, its symptoms and host-range.stalk rot of maize, caused by erwinia carotovora f. sp. zeae sabet (re-designated as pectobacterium chrysanthemi pathovar. zeae by kelman 1974) showed first premature withering and drying up of the uppermost leaves which was soon followed by the lower leaves. the rot either extended from the base upwards (basal rot) or from the top downwards (top rot). in the case of basal rot, the leaves become yellow and the infected tissue becomes brown, soft, and water soaked. internally, the stalk turns int ...1977857508
studies on certain aspects of chemical control of bacterial stalk rot disease of maize.sandoz seed dressing 6335 showing high efficacy in checking the growth of the maize stalk rot pathogen erwinia carotovora f. sp. zeae sabet in culture. brestan, antracol, difolatan, aratan, duter, ceresan wet, flit-406, cuman, blitox-50, streptocycline, agrimycin, terramycin, actidione, aureomycin, chloromycetin, penicillin g, and streptomycin were moderately effective. the rest of the 35 chemicals was negligible in its influence. 15 different chemicals, namely agrimycin, streptocycline, chlorom ...1977857509
microbial conversion of dl-2-amino-delta2-thiazoline-4-carboxylic acid to l-cysteine and l-cystine: screening of microorganisms and identification of products.microorganisms able to form l-cysteine from dl-2-amino-delta2-thiazoline-4-carboxylic acid (dl-atc), a chemical intermediate in the synthesis of dl-cysteine, were isolated from soil samples and classified as pseudomonas sp., pseudomonas cohaerens, p. desmolytica, and p. ovalis. thirteen l-cysteine-producing bacteria were also found in among 463 stock cultures representing 37 genera. these were achromobacter delmarvae. alcaligenes denitrificans, bacillus brevis, brevibacterium flavum, enterobacte ...1977596877
[effect of erwinia carotovora bacteriophage 62 on various indexes of the immune system in animals with ehrlich carcinoma]. 1978567268
physical properties of l-asparaginase from serratia marcescens.purified l-asparaginase from serratia marcescens had an apparent-weight average molecular weight of 171,000 to 180,000 as determined by electrophoresis on polyacrylamide gels and by sedimentation equilibrium at low speed in an analytical ultracentrifuge. a subunit molecular weight of 31,500 +/- 1,500 was estimated for the enzyme after treatment with sodium dodecyl sulfate and urea and electrophoresis on polyacrylamide gels; a similar value was obtained by high-speed sedimentation equilibrium in ...19761107330
trypsin peptide patterns of tryptophan synthase beta2 protein among four species of the enterobacteriaceae.the tryptophan synthase beta 2 protein (ec of escherichia coli, enterobacter aerogenes, serratia marcescens, and erwinia carotovora was purified and compared. two-dimensional total peptide patterns for each of the four beta2 proteins obtained after digestion with trypsin showed that approximately three quarters of the total peptides are common to all four peptides. examination of only arginine-containing peptides showed that approximately half of these peptides are common. from a compa ...1978361723
detection of inhibitors of erwinia carotovora and e. herbicola on thin-layer chromatograms. 19751133145
purification and properties of a third form of anthranilate-5-phosphoribosylpyrophosphate phosphoribosyltransferase from the enterobacteriaceae.anthranilate-5-phosphoribosylpyrophosphate phosphoribosyltransferase was purified from the bacterium erwinia carotovora, a member of the enterobacteriaceae. the enzyme was homogeneous according to the criteria of gel electrophoresis and nh2-terminal amino acid sequence analysis. the molecular weight of the enzyme as determined on a calibrated sephadex g-200 column was 67,000 +/- 2,000. sodium dodecyl sulfate-polyacrylamide gels gave a subunit molecular weight of 40,000 +/- 1,000, suggesting that ...1978338606
application and standardization of various procedures for inoculation of maize by erwinia carotovora f. sp. zeae.hypodermic syringe and toothpick methods were best for evaluating pathogenicity of the maize plant against erwinia carotovora f. sp. zeae, both in glass house and in the field. the portion of the stalk above the ground was best suited for reproducing the disease symptoms. leaf and leaf whorl did not show any rotting hero. seed inoculation showed least mortality percentage. root inoculation was suitable only for young seedlings. cotyledons, as an organ for inoculation for getting best reproducibl ...1977324175
taxonomy of the genus hundred and fifty-six strains of serratia and related bacteria including representatives of enterobacter liquefaciens, enterobacter cloacae, enterobacter aerogenes, erwinia carotovora, erwinia chrysanthemi, erwinia herbicola and erwinia nimipressuralis were studied using 223 morphological, physiological, biochemical and carbon source utilization tests. the results were subjected to computer analysis. at the 80% similarity level all strains, except two, grouped into eight phenons representing ...1977319202
"self-catabolite repression" of pectate lyase in erwinia carotovora.the induction of pectate lyase of erwinia carotovora was repressed by a high concentration of its inducer. the concomitant addition of cyclic adenosine 3',5'-monophosphate reversed this repression.1979217862
effect of three nematicides on the growth of some phytopathogenic bacteria and fungi.the effect of three nematicides, aldicarb, fensulfothion, and phenamiphos at four concentrations (1, 5, 25, and 125 ppm) was tested on the growth of five bacteria, agrobacterium tumefaciens, corynebacterium fascians, erwinia carotovora, pseudomonas solanacearum, and streptomyces scabies and four fungi, fusarium oxysporum f. sp. vasinfectum, fusarium solani, rhizoctonia solani, and sclerotium bataticola. of the bacteria, p. solanacearum was most affected by the chemicals at all concentrations, wh ...1977143857
serologic studies of the maize stalk rot pathogen erwinia carotovora f. sp. zeae.with the help of serological techniques, namely microprecipitin, agglutination, and gel diffusion, sixteen isolates of maize stalk rot pathogen were proved to be identical. serological techniques have thus been utilized as an additional tool for identifying the pathogen erwinia carotovora f. sp. zea. it was also established that the pathogen does not perpetuate in the seed, either externally of internally. it could, however, be found through serological tests that infected ears carried the patho ...197896628
[various properties of virulent and moderate erwinia carotovora phages].a bacterial culture of e. carotovora 8638 lysed by virulent phage 62 after treatment with uv rays produces a moderate phage. the virulent and moderate viruses, antigenically unrealted, have a similar prolonged latent period (100 min) and the period of lysis (60 min). they also differ in the sizes of the capsids and the length of processes which are 750+/-30 and 2000+/-50a in the virulent phage and 600+/-30 and 1500+/-50a, in the moderate phage, respectively. the sedimentation constants are 602 a ...197664030
expression and purification of a soluble tissue factor fusion protein with an epitope for an unusual calcium-dependent antibody.the use of bacterial signal peptides to target recombinant mammalian proteins to the periplasmic space of escherichia coli (to promote proper disulfide bond formation) has met with variable success. we report the design and use of a bacterial expression vector to direct recombinant fusion proteins to the periplasmic space of e. coli: it contains the signal peptide from the pelb gene of erwinia carotovora linked to a small peptide epitope for an unusual calcium-dependent antibody (hpc4). hpc4 bin ...19921283093
immunological relationships of bacterial l-asparaginases.rabbit antisera against l-asparaginase preparations from escherichia coli, erwinia carotovora, citrobacter sp. and chromobacterium violaceum showed on immunoelectrophoresis that only the enzymes from e. coli and citrobacter are immunologically related. purified preparations had to be used to determine the immunological cross-reactions. immunoelectrophoresis at different ph values yielded the zero mobility points of the enzymes. the activity of the er. carotovora preparation was enhanced up to fo ...197654404
bacterial 2,3-butanediol dehydrogenases.enterobacter aerogenes, aeromonas hydrophila, serratia marcescens and staphylococcus aureus possessing l(+)-butanediol dehydrogenase produced mainly meso-butanediol and small amounts of optically active butanediol; acetobacter suboxydans, bacillus polymyxa and erwinia carotovora containing d(-)-butanediol dehydrogenase produced more optically active butanediol than meso-butanediol. resting and growing cells of these organisms oxidezed only one enantiomer of racemic butanediol. the d(-)-butanedio ...197825056
cloning and characterization of the pectate lyase iii gene of erwinia carotovora er.a pectate lyase gene iii (pel iii) of erwinia carotovora er was cloned. the gene was expressed independently of a vector promoter in both e. carotovora er and escherichia coli. the pel iii product was largely excreted in the culture medium of e. carotovora er, while the product was only exported to the periplasmic space and was not excreted in the medium of e. coli. nucleotide sequence analysis of pel iii disclosed an open reading frame of 1,122 bp encoding a protein of 374 amino acids. the dedu ...19911368679
purifications and properties of orotidine-phosphorolyzing enzyme and purine nucleoside phosphorylase from erwinia carotovora aj orotidine-phosphorolyzing enzyme and a purine nucleoside phosphorylase (pnpase) of erwinia carotovora aj 2992, which is a potent producer of ribavirin (1-beta-d-ribofuranosyl-1,2,4-triazole-3-carboxamide), an antiviral agent, from orotidine and 1,2,4-triazole-3-carboxamide (tca), were purified 23-fold and 103-fold, respectively. at each purification step, the orotidine-phosphorolyzing enzyme was always co-purified with an uridine phosphorylase (upase) and its activity could not be separated f ...19911368721
molecular cloning and sequencing of the extracellular pectate lyase ii gene from erwinia carotovora er.erwinia carotovora er produces three extra-cellular pectate lyases (pl i, ii, and iii). the gene for pectate lyase ii (pelii) of e. carotovora er was cloned and expressed both in escherichia coli and e. carotovora er. localization experiments in e. coli showed that pl ii was exclusively in the cytoplasmic space, while pl ii was excreted into the culture medium. the complete nucleotides of the pelii gene were sequenced and found to include one open reading frame of 1122 bp coding for a protein of ...19921369060
polygalacturonic acid trans-eliminase in the osmotic shock fluid of erwinia rubrifaciens: characterization of the purified enzyme and its effect on plant endopolygalacturonic acid trans-eliminase (ec, released by osmotic shock of erwinia rubrifaciens cells, has been purified to near homogeneity (3, 100-fold) by column chromatography on diethylaminoethyl-cellulose, phosphocellulose, and hydroxyapatite-cellulose followed by isoelectric focusing. it has a molecular weight of 41,000, s20,w of 3.09s, an isoelectric point of ph 6.25, ph optimum of 9.5, and a temperature optimum of 37 c and requires ca2+ with an optimum concentration of 0.5 ...19766435
analysis of eight out genes in a cluster required for pectic enzyme secretion by erwinia chrysanthemi: sequence comparison with secretion genes from other gram-negative bacteria.many extracellular proteins produced by erwinia chrysanthemi require the out gene products for transport across the outer membrane. in a previous report (s. y. he, m. lindeberg, a. k. chatterjee, and a. collmer, proc. natl. acad. sci. usa 88:1079-1083, 1991) cosmid pcpp2006, sufficient for secretion of erwinia chrysanthemi extracellular proteins by escherichia coli, was partially sequenced, revealing four out genes sharing high homology with pulh through pulk from klebsiella oxytoca. the nucleot ...19921429461
small molecule-mediated density-dependent control of gene expression in prokaryotes: bioluminescence and the biosynthesis of carbapenem antibiotics.sophisticated signal transduction systems enable prokaryotes to sense their growth environment and mount an appropriate adaptive response. signal transduction and gene regulation through the phosphorylation of two regulatory components is now recognised as one of the major global regulatory networks in bacteria. however, not all types of sensor-regulator circuits relay information via phosphoryl transfer. the vibrio fischeri luxr protein which has previously been characterised as a member of the ...19921478452
nucleotide sequence analysis and comparison of the lexa genes from salmonella typhimurium, erwinia carotovora, pseudomonas aeruginosa and pseudomonas putida.the complete nucleotide sequences of the lexa genes from salmonella typhimurium, erwinia carotovora, pseudomonas aeruginosa and pseudomonas putida were determined; the dna sequences of the lexa genes from these bacteria were 86%, 76%, 61% and 59% similar, respectively, to the escherichia coli k12 gene. the predicted amino acid sequences of the s. typhimurium, e. carotovora and p. putida lexa proteins are 202 residues long whereas that of p. aeruginosa is 204. two putative lexa repressor binding ...19921494343
physicochemical and antigenic properties of the erwinia carotovora bacteriophages. 19752849
high-affinity iron uptake systems present in erwinia carotovora subsp. carotovora include the hydroxamate siderophore aerobactin.the phytopathogenic bacterium erwinia carotovora subsp. carotovora w3c105 produced the hydroxamate siderophore aerobactin under iron-limiting conditions. a survey of 22 diverse strains of e. carotovora revealed that strain w3c105 alone produced aerobactin. the ferric-aerobactin receptor of strain w3c105 was an 80-kda protein, identified by immunoblots of sarkosyl-soluble proteins obtained from e. carotovora cells grown in iron-depleted medium and probed with antiserum raised against the 74-kda f ...19921569027
[the influence of choramphenicol and streptomycinsulphate on the growth of pectobacterium carotovorum var. atrosepticum (van hall) dowson (author's transl)].the influence of the antibiotics chloraphenicol (d-threo-form) and streptomycin sulphate against pectobacterium carotovorum var. atrosepticum (syn. erwinia carotovora) was investigated in vitro and on potato tubers. chloramphenicol showed a greater effect as streptomycinsulphate in the applied concentrations. the success of the control of soft rot is determined by the period between the inoculation and the treatment of the tubers. 6 several strains of p. carotovorum var. atrosepticum showed a di ...19761037182
sensitivity distribution of phytopathogenic bacteria and fungi to antibiotics.the minimal inhibitory concentrations (mic) of various antibiotics and fungicides for erwinia carotovora, pseudomonas coronafaciens var. atropurpurea, p. lachrymans, alternaria mali, a. kikuchiana, pyricularia oryzae, botrytis sp. and sclerotinia sp. isolated from diseased plants in various localities of japan were examined to enable the isolates to be gruoped into sensitive and resistant strains. to minimize the effects of various variable conditions, mic of isolates were pooled for either 2 or ...19761033174
characterization of the erwinia carotovora peh gene and its product polygalacturonase.the peh gene, encoding polygalacturonase (peh), was identified in erwinia carotovora strain ec and cloned in escherichia coli. recombinant peh (re-peh) was purified from e. coli strain 706 containing peh on a recombinant plasmid. the activity of the re-peh protein is optimal at ph 5.5. the n-terminal and internal amino acid (aa) sequences of re-peh were determined and compared to the aa sequence deduced from the nucleotide (nt) sequence of the cloned peh. the re-peh has no similarity, based on e ...19921644302
an investigation of the electronic and steric environments of tyrosyl residues in ribonuclease a and erwinia carotovora l-asparaginase through fluorescence quenching by caesium, iodide and phosphate ions.the fluorescence lifetimes and relative quantum yields of several derivatives of tyrosine are reported. the quenching of the fluorescence of these compounds by phosphate, caesium and iodide ions has been investigated; the encounter rate constants, calculated from the quenching parameters and lifetimes, show a clear dependence on the charges borne by the quenchers and fluorophores. the ratio of the stern-volmer constants of iodide and caesium, ions of similar size, defines an electrostatic parame ...1976986170
nucleotide sequence and molecular characterization of pnla, the structural gene for damage-inducible pectin lyase of erwinia carotovora subsp. carotovora a previous study, pnla (the dna damage-inducible structural gene for pectin lyase) of erwinia carotovora subsp. carotovora 71 was localized to a 1.4-kb dna segment within a 3.4-kb ecori fragment (j. l. mcevoy, h. murata, and a. k. chatterjee, j. bacteriol. 172:3284-3289, 1990). we present here dna sequence data for a 2.2-kb region revealing an open reading frame of 870 bases, corresponding to a protein (pnl) of an approximate molecular mass of 32,100 da and an isoelectric point of 9.92. altho ...19911705542
characterization of a polygalacturonase gene of aspergillus niger rh5344.we have cloned a gene encoding a polygalacturonase (pg) in the filamentous fungus aspergillus niger rh5344. the structural gene comprises 1141 bp coding for 362 amino acids and the open reading frame is disrupted by one intron of 52 bp. eukaryotic consensus sequences for transcription regulation are found only in deviated forms. the biological functionality of the isolated pg gene was established by retransformation in a. niger and aspergillus awamori. in addition, we have found that the pg prot ...19911787790
l-asparaginase--a reassessment.the authors review some historical data connected with the discovery of the enzyme l-asparaginase and then summarize the basic informations concerning the mechanism of action, clinical application, toxicity and future prospectives of this substance. from the clinical viewpoint, a broader use of l-asparaginase prepared from erwinia carotovora instead of escherichia coli seems to be warranted.19911837674
the modification of tyrosyl residues in l-asparaginases and ribonuclease a with fremy's salt. implication of tyrosyl residues in the activity of l-asparaginases from e. coli b and erwinia carotovora. 1975776662
proteins, phospholipid distribution and fluidity in membranes of the gram-negative bacterium erwinia carotovora [proceedings]. 1978744424
erwinia carotovora subsp. carotovora extracellular protease: characterization and nucleotide sequence of the gene.the prt1 gene encoding extracellular protease from erwinia carotovora subsp. carotovora ec14 in cosmid pca7 was subcloned to create plasmid psk1. the partial nucleotide sequence of the insert in psk1 (1,878 bp) revealed a 1,041-bp open reading frame (orf1) that correlated with protease activity in deletion mutants. orf1 encodes a polypeptide of 347 amino acids with a calculated molecular mass of 38,826 da. escherichia coli transformed with psk1 or psk23, a subclone of psk1, produces a protease ( ...19911917878
one-step cloning system for isolation of bacterial lexa-like genes.a system to isolate lexa-like genes of bacteria directly was developed. it is based upon the fact that the presence of a lexa(def) mutation is lethal to sula+ cells of escherichia coli. this system is composed of a sula- lexa(def) hsdr- strain and a lexa-conditional killer vector (plasmid pua165) carrying the wild-type sula gene of e. coli and a polylinker in which foreign dna may be inserted. by using this method, the lexa-like genes of salmonella typhimurium, erwinia carotovora, pseudomonas ae ...19911938926
cloned erwinia chrysanthemi out genes enable escherichia coli to selectively secrete a diverse family of heterologous proteins to its milieu.the out genes of the enterobacterial plant pathogen erwinia chrysanthemi are responsible for the efficient extracellular secretion of multiple plant cell wall-degrading enzymes, including four isozymes of pectate lyase, exo-poly-alpha-d-galacturonosidase, pectin methylesterase, and cellulase. out- mutants of er. chrysanthemi are unable to export any of these proteins beyond the periplasm and are severely reduced in virulence. we have cloned out genes from er. chrysanthemi in the stable, low-copy ...19911992458
nucleotide sequence of pnl gene from erwinia carotovora er.the nucleotide sequence of pnl gene encoding pectin lyase (pnl; ec4.2.2.10)from erwinia carotovora er was determined. the structural gene of pnl consisted of 942 base pairs. an open reading frame that could encode a 33,700 dalton polypeptide consisting 314 amino acids was assigned. the molecular size of the polypeptide predicted from the amino acid composition was close to the value of pnl determined in e.carotovora er. the nucleotide sequence of the 5'-flanking region showed the presence of the ...19912018526
molecular cloning of omprs, a regulatory locus controlling production of outer membrane proteins in erwinia carotovora subsp. carotovora.a locus, omprs, controlling synthesis of outer membrane proteins was cloned from erwinia carotovora subsp. carotovora (ecc) by complementation of an escherichia coli ompr-envz mutant. the ecc omprs locus was both structurally and functionally similar to the ompr-envz operon controlling porin gene expression in e. coli as shown by dna hybridization and complementation of e. coli ompr and envz mutants. furthermore, introduction of omprs into e. coli delta (ompr-envz) strains restored the osmoregul ...19912038301
[plasmid r68.45 and s-a transduction by the temperate bacteriophage 59 of erwinia carotovora 268].temperature bacteriophage 59 of erwinia carotovera 268 had transduced extrachromosomal dna: plasmids of r68.45 and s-a. before plasmid transduction experiments the suitable donor strains of indicator culture erwinia horticola 450 harbouring r68.45 and s-a were created. the frequency of plasmid r68.45 transfer from pseudomonas putida to e. horticola 450-8 by conjugation was equal to 5 x 10(-8) per a donor cell and in the case of s-a--from e. coli c600 for the same recipient cells--was 2 x 10(-6). ...19912067419
facile isolation of endo-pectate lyase from erwinia carotovora based on electrostatic interaction.endo-pectate lyase (pate) from erwinia carotovora was selectively cosedimented with extracellularly produced lipopolysaccharide-lipid complex (lpslc) through dialysis of the cell free culture broth. the selective isolation of pate was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. the cosedimentation of the pate with lpslc was initiated by decreasing conductivity of the solution and terminated at approx 1 m siemens (mscm-1). as much as 62% of pate activity in the culture ...19902091529
[polyamines in representative taxa of coryneform and other bacteria].the composition of polyamines is studied for the first time in representatives of the genus micrococcus and taxon "conglomeratus", strains staphylococcus aureus ccm 209, deinococcus erythromyxa ccm 706 as well as of erwinia carotovora atcc 15713 polyamines, which are not extracted by perchloric acid. considerable amounts of spermine and rarely of spermidine are revealed in cells of gram positive microorganisms, that differs them from gram negative bacteria possessing high concentrations of putre ...19902115965
[purification of microbial asparaginases by the aid of affinity chromatography].asparaginases from escherichia coli and erwinia caratovora were isolated and purified by column chromatography with a specific sorbent (sepharose, covalently bound to n-alpha-(6-aminohexyl)-d-asparagine). homogenous asparaginase from e. coli was isolated by one-step procedure, while the enzyme from er. carotovara was 50-60-fold purified. asparaginase from mycobacterium n. sp. was found not to bind with the specific sorbent.19751095078
characterization of transposon insertion out- mutants of erwinia carotovora subsp. carotovora defective in enzyme export and of a dna segment that complements out mutations in e. carotovora subsp. carotovora, e. carotovora subsp. atroseptica, and erwinia chrysanthemi.soft-rotting erwinia spp. export degradative enzymes to the cell exterior (out+), a process contributing to their ability to macerate plant tissues. transposon (tn5, tn10, tn10-lacz) insertion out- mutants were obtained in erwinia carotovora subsp. carotovora 71 by using plasmid and bacteriophage lambda delivery systems. in these mutants, pectate lyases, polygalacturonase, and cellulase, which are normally excreted into the growth medium, accumulated in the periplasm. however, localization of th ...19902160934
[erwinia carotovora as a recipient system for the natural hybrid plasmid rp4::mu cts62].the plasmid rp4::mu cts62 is transferred from escherichia coli cells into a recipient strain erwinia carotovora 268 by conjugation with the frequency 1.5-5 x 10(-7) per donor cell. the maximal frequencies of transfer are obtained by cultivation of donor and recipient cells for 3-5 h on the filters. structural and functional validity of the plasmid in transconjugants is expressed in preservation of all antibiotic-resistant markers of rp4, thermosensitivity to growth at 42 degrees c as well as spo ...19902175013
cloning and expression of pectin lyase gene from erwinia carotovora in escherichia coli.a pectin lyase (pnl; ec gene of erwinia carotovora er was cloned and expressed in escherichia coli. the analysis of the nucleotide sequence of the 0.6 kb stui-ecori fragment, which was hybridized with the mixed oligonucleotide probe for pnl gene, revealed the presence of an open reading frame (0rf) and correlated exactly with the known n-terminal 18 amino acid sequence of pnl. when a plasmid ptn2159, which has a bamhi-ecori fragment containing this orf, was introduced into e. coli jm10 ...19902185758
a family of positive regulators related to the pseudomonas putida tol plasmid xyls and the escherichia coli arac activators.the xyls family consists of a least 8 different transcriptional regulators. six of these proteins are positive regulators for the catabolism of carbon sources (benzoate and sugars) in escherichia coli, pseudomonas putida and erwinia carotovora, and two of them are involved in pathogenesis in escherichia coli and yersinia enterocolitica. based on protein alignments, the members of this family exhibit a long stretch of homology at the c-terminal end. the regulators involved in the catabolism of ca ...19902186376
molecular cloning and characterization of an erwinia carotovora subsp. carotovora pectin lyase gene that responds to dna-damaging agents.reca-mediated production of pectin lyase (pnl) and the bacteriocin carotovoricin occurs in erwinia carotovora subsp. carotovora 71 when this organism is subjected to agents that damage or inhibit the synthesis of dna. the structural gene pnla was isolated from a strain 71 cosmid gene library following mobilization of the cosmids into a moderate pnl producer, strain 193. the cosmid complemented pnl::tn5 but not ctv::tn5 mutations. a constitutive level of pnl activity was detected in reca+ and rec ...19902188956
[erwinia carotovora in the lupine rhizosphere]. 19751214636
structural analysis of the peha gene and characterization of its protein product, endopolygalacturonase, of erwinia carotovora subspecies carotovora.a clone producing a polygalacturonase (ec in escherichia coli was isolated from a genomic library of erwinia carotovora subspecies carotovora constructed in puc18. the dna segment carrying the corresponding structural gene, named peha, contained an open reading frame (orf) encoding a 402-amino-acid (aa) polypeptide with an mr of 42,849. in e. carotovora the polygalacturonase was synthesized with a 26-aa cleavable signal peptide. the mature 376-aa peha had a calculated mr of 40,064 and ...19902215212
homology among bacterial catalase genes.catalase activities in crude extracts of exponential and stationary phase cultures of various bacteria were visualized following gel electrophoresis for comparison with the enzymes from escherichia coli. citrobacter freundii, edwardsiella tarda, enterobacter aerogenes, klebsiella pneumoniae, and salmonella typhimurium exhibited patterns of catalase activity similar to e. coli, including bifunctional hpi-like bands and a monofunctional hpii-like band. proteus mirabilis, erwinia carotovora, and se ...19902253114
[cloning and expression of clostridium thermocellum f7 endoglucanase gene in gram-negative bacteria].the endoglucanase gene of clostridium thermocellum f7 was cloned in escherichia coli cells using pkm4 vector. both the physical mapping and analysis of the gene products in the e. coli mini-cells system suggest cloning of a new cel gene different from those described earlier. the activity of endoglucanase in e. coli cells is localized in the periplasm, which correlates with secretion of enzymes of this type in c. thermicellum. apart from 2 major components with mr 42.5 and 43 kda, corresponding ...19902258032
dna sequence analysis of the reca genes from proteus vulgaris, erwinia carotovora, shigella flexneri and escherichia coli b/r.the complete nucleotide sequences of the reca genes from escherichia coli b/r, shigella flexneri, erwinia carotovora and proteus vulgaris were determined. the dna sequence of the coding region of the e. coli b/r gene contained a single nucleotide change compared with the e. coli k12 gene sequence whereas the s. flexneri gene differed at 7 residues. in both cases, the predicted proteins were identical in primary structure to the e. coli k12 reca protein. the dna sequences of the reca genes from e ...19902274037
[mapping of the c-region of the temperate phage 59 of erwinia carotovora 268].the present communication deals with the genetic mapping of the temperate bacteriophage 59 immunity region. morphological mutants of the c-type of the temperate bacteriophage 59 were isolated after uv-treatment of the phage. complementation analysis of 36 c-mutants has shown that the bacteriophage 59 c-region consists of at least four genes responsible for the establishment of lysogenization. the two-factor crosses were used to determine the mutual positions (order and distances) of the c-mutati ...19902362599
cels: a novel endoglucanase identified from erwinia carotovora subsp. carotovora.a plasmid clone expressing a beta(1,4)-glucan glucanohydrolase (ec; endoglucanase) in escherichia coli was isolated from a genomic library of erwinia carotovora subsp. carotovora. the dna segment carrying the corresponding structural gene, named cels, contained an open reading frame encoding a 264-amino acid (aa) polypeptide. the n-terminal aa sequence of cels showed that the protein was synthesized with a 32-aa cleavable signal peptide. the mature 232-aa cels had a calculated mr of 26,2 ...19902379837
enumerating low densities of genetically engineered erwinia carotovora in inexpensive, quantitative, and sensitive technique was developed for detection of genetically engineered erwinia carotovora in soil samples. enrichment media, antibiotic resistance, and most probable number (mpn) analysis were used to enumerate as few as 1 to 10 target cells/10 g soil. the mpn technique recovered significantly higher cell densities than plating; however, densities estimated by the two techniques were strongly correlated. after inoculation of soil microcosms with genetically e ...19901366379
[mu-induced deletions and mutations of erwinia carotovora chromosomes, including resident prophages e105 and 59].the plasmid rp4::mu cts62 in stably inherited by erwinia carotovora 268 strain. under the conditions of thermoinduction bacteriophage mu is segregated and completely eliminated more intensively than in escherichia coli cells. at thermoinduction the transposition of bacteriophage mu cts62 into different chromosomal sites takes place, causing the induction of chlorate resistant and auxotrophic mutants with the frequency of 10(-4). two clones deficient in production of 2 of the 4 resident prophages ...19921406759
genetic evidence for an activator required for induction of pectin lyase in erwinia carotovora subsp. carotovora by dna-damaging erwinia carotovora subsp. carotovora 71, the induction of pectin lyase (pnl), the bacteriocin carotovoricin (ctv), and cellular lysis (lss) requires a reca function. we obtained mutants wherein a pleiotropic defect, i.e., the lack of induction with mitomycin c, is not restored by the reca+ dna. from a genomic library of strain 71, a cosmid (pakc280) that restored induction of pnl, ctv, and lss by mitomycin c was isolated. the activator function, designated rdg for regulator of damage-inducibl ...19921644776
isolation and characterisation of transposon-induced mutants of erwinia carotovora subsp. atroseptica exhibiting reduced virulence.the blackleg pathogen erwinia carotovora subsp. atroseptica (eca) causes an economically important disease of potatoes. we selected a genetically amenable eca strain for the genetic analysis of virulence. tn5 mutagenesis was used to generate nine mutants which exhibited reduced virulence (rvi-) of strain scri1043. following physiological characterisation, mutants were divided into three classes: (1) auxotrophs; (2) extracellular enzyme mutants; and (3) a growth rate mutant. the isolation of thes ...19892549365
purification of rna polymerase and transcription-termination factor rho from erwinia carotovora.erwinia carotovora rna polymerase consists of the holoenzyme structure sigma 2 beta beta' sigma as found in escherichia coli and other bacteria. e. carotovora rna polymerase can synthesize rna using lambda, t7 of t4 dna as templates; however, it is two times less active on these templates and is more temperature-sensitive than the e. coli enzyme. the alpha subunit of the e.. carotovora enzyme is lower in molecular mass than its e. coli counterpart. the sigma factors from e. coli and e. carotovor ...19852578393
differential activation of potato 3-hydroxy-3-methylglutaryl coenzyme a reductase genes by wounding and pathogen challenge.potato genes encoding 3-hydroxy-3-methylglutaryl coenzyme a reductase (hmgr) were expressed in response to pathogen, elicitor, and wounding. hmgr catalyzes the rate-limiting step in isoprenoid biosynthesis leading to accumulation of phytoalexins and steroid glycoalkaloids. wounding caused increases in hmgr mrna levels. a rapid and transient peak occurred 30 minutes after wounding, followed by a slower peak at 14 hours; both were correlated with increased enzyme activity. induction of hmgr mrna b ...19911840919
attenuation of asparaginase-induced hyperglycemia after substitution of the erwinia carotovora for the escherichia coli enzyme preparation.l-asparaginase, an enzyme with established antileukemic activity, increases the induction rate and duration of remission of acute lymphoblastic leukemia when added to vincristine and prednisone for induction therapy. enzymes derived from two different bacterial sources (escherichia coli and erwinia carotovora) are in common use. these enzymes may be associated with toxic reactions of differing frequency and severity. specifically, the complication of enzyme-induced hyperglycemia may be seen more ...19892643458
extracellular and periplasmic isoenzymes of pectate lyase from erwinia carotovora subspecies carotovora belong to different gene families.pectate lyase (pel) plays a crucial role in the maceration of vegetables by soft rot erwinia spp. we have characterized the four pel isoenzymes of erwinia carotovora subspecies carotovora strain scri193. in this paper we concentrate on two isoenzymes which have different locations in scri193: plb is periplasmic and plc is extracellular. comparison of the gene products and nucleotide sequences of pelb and pelc allowed us to assign them to different gene families. in addition, we have identified a ...19892695748
[an electron microscopic study of the structure of erwinia carotovora 268 bacteria].the electron-microscopic studies have shown that the bacterium erwinia carotovora 268 (a producer of l-asparaginase) does not differ essentially from other representatives of enterobacteria. when growing on the artificial nutrient medium m9, the division of bacteria is inhibited and the bacterial cell itself elongates. certain functionally specified structural peculiarities such as the formation of the protein microcapsule, formation of cavities and compacted sites of the cytoplasm and mucus on ...19892695793
[effect of calcium and magnesium ions on growth and cell division of erwinia carotovora]. 1976969578
[adsorption and reproduction of bacteriophages in erwinia carotovora]. 1976948268
[effect of a uniform magnetic field on erwinia carotovora bacteriophage 62]. 1977865310
identification of erwinia sp., causing stalk rot of maize in egypt.the physiological characteristics, pathogenic propensities, and the sensitivity towards certain antibiotics of the causal bacterium of stalk rot of maize, isolated in egypt, were studied. different isolates of erwinia carotovora and e. atroseptica were included for comparative studies. the antibiotic sensitivity tests (including erythromycin) are of no value in differentiating between e. carotovora and e. atroseptica. the cultures of the causal bacterium of stalk rot of maize showed physiologica ...1978749412
use of tnphoa to enrich for extracellular enzyme mutants of erwinia carotovora subspecies carotovora.soft rot erwinia species secrete a range of enzymes into the extracellular environment. therefore, the genetically amenable erwinia system is a useful model for the study of protein secretion by gram-negative bacteria. we have used a lambda-sensitive derivative of erwinia carotovora subspecies carotovora (ecc) and the transposon tnphoa, to isolate a range of extracellular enzyme mutants. the use of tnphoa provides an enrichment for extracellular enzyme mutants over other transposon-based systems ...19872834624
[restriction map of the temperate phage e105 from the polylysogenic culture of erwinia carotovora 268].dna structure of the temperate bacteriophage e105 from polylysogenic culture of erwinia carotovora 268 has been studied. the viral 29.12-29.17 md dna has been shown to be linear and nonpermuted. the complete restriction map of the viral dna has been constructed for mvai and hpai and partial for eco31 restriction endonucleases based on the pair hydrolysis of the native dna as well as its fragments. altogether, 19 sites for restriction endonucleases have been localized on bacteriophage dna.19882841593
characterization of type 1 and mannose-resistant fimbriae of erwinia spp.type 1 fimbriae from erwinia carotovora subsp. carotovora and mannose-resistant fimbriae from erwinia rhapontici were purified and characterized. the type 1 fimbrillin had an apparent molecular weight of 16,500; that of the mannose-resistant fimbrillin was 18,000. the amino-terminal amino acid sequences of the two fimbrillins were related, but tryptic peptide maps showed significant differences between the proteins. no serological cross-reaction was found between the two fimbrial filaments, nor ...19872883172
[restriction map of permuted dna of erwinia carotovora temperate bacteriophage 59].the cyclic permutation and terminal redundancy were found in the genomes of erwinia carotovora temperate bacteriophage 59 by electron microscopic studies. the headful mechanism for bacteriophage dna cleavage and packaging during the phage morphogenesis was confirmed by the restriction analysis technique. restriction map of the bacteriophage 59 dna was constructed for restriction endonucleases bamhi, bg1ii, eco31, sa1i, smai, ecori.19882895890
cdna cloning of carrot extracellular beta-fructosidase and its expression in response to wounding and bacterial infection.we isolated a full-length cdna for apoplastic (extracellular or cell wall-bound) beta-fructosidase (invertase), determined its nucleotide sequence, and used it as a probe to measure changes in mrna as a result of wounding of carrot storage roots and infection of carrot plants with the bacterial pathogen erwinia carotovora. the derived amino acid sequence of extracellular beta-fructosidase shows that it is a basic protein (pl 9.9) with a signal sequence for entry into the endoplasmic reticulum an ...19902152110
the arac gene of citrobacter freundii.the arac gene of citrobacter freundii was cloned into plasmid pbr322 and expressed in escherichia coli and salmonella typhimurium. the nucleotide sequence and the predicted translational product were determined and compared to those of e. coli, s. typhimurium and erwinia carotovora. the predicted translational product is 281 amino acids (aa) long, identical in size to that of s. typhimurium, and is 11 and 29 aa shorter than that of e. coli and e. carotovora, respectively. the nucleotide sequence ...19872965663
an improved dna sequencing strategy.a modification of hong's systematic dna sequencing strategy is described. the original procedure has been simplified and transfectant yield increased. after dnase i limited cleavage in the presence of mn2+, the single-cut linear dna does not have to be separated from supercoiled or open circular dna on an agarose gel. after ligation, the dna is digested with a second restriction endonuclease for which a unique cleavage site resides between the insert and the first restriction endonuclease cuttin ...19852992313
pulb113, an rp4::mini-mu plasmid, mediates chromosomal mobilization and r-prime formation in erwinia amylovora, erwinia chrysanthemi, and subspecies of erwinia carotovora.the rp4::mini-mu plasmid pulb113, transferred from escherichia coli strain mxr, was stable and transfer proficient in erwinia amylovora strain ea303, e. carotovora subsp. atroseptica strain eca12, e. carotovora subsp. carotovora strain ecc193, and e. chrysanthemi strain ec183. the plasmid mobilized an array of erwinia sp. chromosomal markers (e. amylovora: his+,ilv+,rbs+,ser+,thr+;e. chrysanthemi:arg+,his+,ilv+,leu+; e. carotovora subsp. atroseptica: arg+,gua+,leu+,lys+,pur+,trp+; e. carotovora ...19852992373
effects of a mutation that eliminates udp glucose-pyrophosphorylase on the pathogenicity of erwinia carotovora subsp. carotovora.a nonpathogenic mutant of erwinia carotovora obtained by mu d1 mutagenesis was defective in the ability to utilize several carbon sources. the basis of the mutation was analyzed biochemically and shown to be a defect in the ability to form udp glucose-pyrophosphorylase. the nonpathogenic phenotype of the mutant was caused by its sensitivity to galactose.19852995320
effect of dusts emitted by copper smelters on erwinia carotovara.the effect of the dust emitted by copper smelters on erwinia carotovora was examined. the dust contains considerable amounts of heavy metals which inhibited the growth and enzymatic activity of the bacterial cultures. the inhibition of dehydrogenase and protease activity was greater than that of the growth rate. the phenolase production and virulence of strain was also inhibited, depending on the doses of dust. calcium carbonate counteracted the toxic effect of dust, restoring enzymatic activity ...1977576108
isolation of maltotetraose from streptomyces as an antibiotic against erwinia carotovora. 19751112768
requirement for two or more erwinia carotovora subsp. carotovora pectolytic gene products for maceration of potato tuber tissue by escherichia coli.several genes encoding enzymes capable of degrading plant cell wall components have been cloned from erwinia carotovora subsp. carotovora ec14. plasmids containing cloned ec14 dna mediate the production of endo-pectate lyases, exo-pectate lyase, endo-polygalacturonase, and cellulase(s). escherichia coli strains containing one of these plasmids or combinations of two plasmids were tested for their ability to macerate potato tuber slices. only one e. coli strain, containing two plasmids that encod ...19863013836
characterization of the erwinia carotovora pelb gene and its product pectate lyase.the pelb gene encodes pectate lyase b, one of three pectate lyases identified in erwinia carotovora ec. pectate lyase b was purified from escherichia coli containing the pelb gene on a recombinant plasmid. the activity of the protein was optimal at a ph of 8.3. the amino acid composition, n-terminal amino acid sequence, and c-terminal peptide sequence were determined and compared with the polypeptide sequence deduced from the dna sequence of pelb. purified pectate lyase b started at amino acid 2 ...19873040692
the post-harvest fruit rots of tomato (lycopersicum esculentum) in nigeria.a survey of the post-harvest fruit rot diseases of tomato was conducted in five states of nigeria. during severe infections, the diseases could cause 25% loss at harvest and 34% loss of the remaining product in transit, storage and market stalls; thus giving an overall loss of about 50% of the product. two types of rots, soft and dry were recognised. the soft rot was found to account for about 85% and the dry rot about 15% of the overall loss. erwinia carotovora, rhizopus oryzae, r. stolonifer, ...1979471028
preparation and properties of membranes from the gram-negative bacterium erwinia carotovora. 19751193287
biosynthetic utilization of ethanolamine by erwinia carotovora. 19751193291
effect on microorganisms of volatile compounds released from germinating seeds.volatile compounds evolved from germinating seeds of slash pine, bean, cabbage, corn, cucumber, and pea were evaluated for their ability to support growth of microorganisms in liquid mineral salts media lacking a carbon source. growth of eight bacteria was measured turbidimetrically and of six fungi as dry weight of mycelium. volatiles caused increased growth of pseudomonas fluorescens, bacillus cereus, erwinia carotovora, agrobacterium tumefaciens, a. radiobacter, rhizobium japonicum, mucor muc ...19751201509
microorganisms associated with mouldiness of dried yam chips and their prevention.the broad objective of this study was to isolate and identify the microorganisms causing mouldiness of stored yam chips and to look for ways of preventing the problem. microorganisms isolated included aspergillus flavus, a. glaucus, a. nidulans, a. niger, a. ochraceous, a. tamarii, a. candidus, penicillium oxalicum, trichoderma longibrachyatum, rhizopus nigricans, cylindrocarpon radicicola, neurospora crassa, botryodiplodia theobromae, bacillus subtilis, bacillus cereus, erwinia carotovora and s ...19883231261
bacteriophage t4 resistant mutants of the plant pathogen erwinia carotovora.bacterial lipopolysaccharide (lps) has been implied in a variety of pathogenic and symbiotic plant-bacterium interactions. in order to study the role of lps in pathogenicity of erwinia carotovora, a broad host range phytopathogen, we isolated lps defective mutants of two subspecies of erwinia carotovora, subsp. carotovora (ecc) and subsp. astroseptica (eca). this was accomplished by using the escherichia coli phage t4 as a selective agent. screening of erwinia isolates revealed that some of them ...19883241545
conservation of an atp-binding domain among reca proteins from proteus vulgaris, erwinia carotovora, shigella flexneri, and escherichia coli k-12 and b/r.the purified reca proteins encoded by the cloned genes from proteus vulgaris, erwinia carotovora, shigella flexneri, and escherichia coli b/r were compared with the reca protein from e. coli k-12. each of the proteins hydrolyzed atp in the presence of single-stranded dna, and each was covalently modified with the photoaffinity atp analog 8-azidoadenosine 5'-triphosphate (8n3atp). two-dimensional tryptic maps of the four heterologous reca proteins demonstrated considerable structural conservation ...19883286605
biosynthesis of maltotetraose as an antibiotic against erwinia carotovora by a streptomyces amylase. 19751201973
microbial coagulation of alfalfa green juice.of the different bacterial strains isolated from alfalfa raw material, nine were able to coagulate the protein fraction of alfalfa green juice. the two strains showing the highest efficiency were further used for coagulation experiments. they were classified as erwinia carotovora and escherichia coli. juice samples inoculated (1:10 to 1:100) with stationary-phase cultures were efficiently coagulated. the amount of protein recovered was equivalent to that obtained when the juice was heat treated. ...19873314708
[ultrastructure and chemical composition of lipopolysaccharides of erwinia carotovora. ii. influence of the extraction method]. 19751234622
characterization of the erwinia carotovora pela gene and its product pectate lyase a.the pela gene from erwinia carotovora strain ec encodes pectate lyase a (pla). the gene was cloned and sequenced. the pla was purified from escherichia coli containing the pela gene on a recombinant plasmid. the optimum ph for the enzyme reaction is at ph 8.5; the optimum temperature for the enzyme reaction is at 40 degrees c; and the purified pla alone can macerate potato slices. nucleotide sequence of pela and the deduced polypeptide sequence of pla were compared with the pelb gene of e. carot ...19883371662
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