Publications

the corynebacterium xerosis composite transposon tn5432 consists of two identical insertion sequences, designated is1249, flanking the erythromycin resistance gene ermcx.	analysis of the 50-kb r-plasmid ptp10 from the clinical isolate corynebacterium xerosis m82b revealed that the erythromycin resistance gene, ermcx, is located on a 4524-bp composite transposable element, tn5432. the ends of tn5432 are identical, direct repeats of an insertion sequence, designated is1249, encoding a putative transposase of the is256 family. is1249 consists of 1385 bp with 45/42 imperfect terminal inverted repeats. the nucleotide sequence of the 1754-bp tn5432 central region is 99 ...	1995	8559800
prophage distribution in coryneform bacteria.	four temperate bacteriophages of corynebacteria were isolated after uv induction. phages phi 304l and phi 304s were both induced from corynebacterium glutamicum atcc 13058, atcc 21488, atcc 21649 and atcc 21650 strains, and have no known sensitive host. phages phi 15 and phi 16 were both induced from atcc 14020 and atcc 21792. phage phi 15 formed turbid plaques on corynebacterium sp. atcc 21857 and on c. glutamicum atcc 13058, atcc 21488, atcc 21649 and atcc 21650. phage phi 16 produced turbid p ...	1995	8525066
molecular cloning of the hom-thrc-thrb cluster from bacillus sp. ulm1: expression of the thrc gene in escherichia coli and corynebacteria, and evolutionary relationships of the threonine genes.	a 6.5 kb dna fragment containing the gene (thrc) encoding threonine synthase, the last enzyme of the threonine biosynthetic pathway, has been cloned from the dna of bacillus sp. ulm1 by complementation of escherichia coli and brevibacterium lactofermentum thrc auxotrophs. complementation studies showed that the thrb gene (encoding homoserine kinase) is found downstream from the thrc gene, and analysis of nucleotide sequences indicated that the hom gene (encoding homoserine dehydrogenase) is loca ...	1995	8768250
isolation of insertion elements from gram-positive brevibacterium, corynebacterium and rhodococcus strains using the bacillus subtilis sacb gene as a positive selection marker.	the sacb gene of bacillus subtilis was successfully applied in various arthrobacter, brevibacterium, corynebacterium and rhodococcus strains for the isolation of transposable elements. three different insertion sequence (is) elements entrapped in sacb were isolated. the is elements is-bl and is-cg isolated from brevibacterium lactofermentum and corynebacterium glutamicum, respectively, were found to be similar in size (1.45 kb) and generated target duplications of 8 bp. their inverted repeats sh ...	1995	7896070
structure of the gluabcd cluster encoding the glutamate uptake system of corynebacterium glutamicum.	to assess the mechanism and function of the glutamate uptake system of gram-positive corynebacterium glutamicum, a mutant deficient in glutamate uptake was isolated and was then used to isolate a dna fragment restoring this deficiency. in a low-copy-number vector, this fragment resulted in an increased glutamate uptake rate of 4.9 nmol/min/mg (wild type, 1.5 nmol/min/mg). in addition, carbon source-dependent regulation of the glutamate uptake system was determined with the fragment, showing that ...	1995	7868586
fermentation and recovery of glutamic acid from palm waste hydrolysate by ion-exchange resin column.	glutamic acid produced from palm waste hydrolysate by fermentation with brevibacterium lactofermentum atcc 13869 is produced with a remarkably high yield compared with that produced from pure glucose as a carbon source. the produce yield is 70 g/l with glucose, wherease, when palm waste hydrolysate is the fermentation medium in the same bioreactor under same conditions, it is 88 g/l. the higher yield may be attributed to the fact that this organism has the ability to convert sugars other than on ...	1995	18623521
studies on urea biosensors based on immobilized corynebacterium glutamicum and their kinetic response processes.	two novel biosensors for urea based on immobilized corynebacterium glutamicum 617 and corynebacterium glutamicum atcc13032 in calcium alginate gel coupled with an ammonia gas-sensing electrode, were designed and constructed. calibration plots of measured potential difference (mv) vs. log of urea concentration were linear in the range of 5.6 x 10(-5)-1.4 x 10(-2) and 5.6 x 10(-5)-1.1 x 10(-2) mol l(-1), with slopes of 59.2 and 61.3 mv per decade respectively, in ph 8.0, 0.1 mol l(-1) phosphate bu ...	1995	18966389
use of feedback-resistant threonine dehydratases of corynebacterium glutamicum to increase carbon flux towards l-isoleucine.	the biosynthesis of l-isoleucine proceeds via a highly regulated reaction sequence connected with l-lysine and l-threonine synthesis. using defined genetic corynebacterium glutamicum strains characterized by different fluxes through the homoserine dehydrogenase reaction, we analyzed the influence of four different ilva alleles (encoding threonine dehydratase) in vectors with two different copy numbers on the total flux towards l-isoleucine. for this purpose, 18 different strains were constructed ...	1995	16535185
triggering glutamate excretion in corynebacterium glutamicum by modulating the membrane state with local anesthetics and osmotic gradients.	corynebacterium glutamicum can be triggered to excrete glutamate by the addition of local anesthetics, particularly tetracaine. glutamate efflux is a carrier-mediated process and not due to unspecific membrane permeabilization. the concentration of local anesthetics triggering optimum excretion depended on the type of anesthetic and varied, ranging from 0.1 (chlorpromazine), 1.3 (tetracaine), and 2.6 mm (butacaine) to 15 mm (benzocaine), in close resemblance to the order of efficiency in anesthe ...	1995	16535186
l-isoleucine production with corynebacterium glutamicum: further flux increase and limitation of export.	the synthesis of l-isoleucine with corynebacterium glutamicum involves 11 reaction steps, in at least five of which activity or expression is regulated. we used four genes and alleles encoding feedback-resistant enzymes (fbr) in various combinations to assay flux increase through the sequence. during strain construction, the order of genes overexpressed was important. only when ilva(fbr) was first overexpressed could hom(fbr) be introduced. this succession apparently prevents the toxic accumulat ...	1996	16535457
growth rate-dependent modulation of carbon flux through central metabolism and the kinetic consequences for glucose-limited chemostat cultures of corynebacterium glutamicum.	the physiological behavior of corynebacterium glutamicum in glucose-limited chemostat cultures was examined from both growth kinetics and enzymatic viewpoints. metabolic fluxes within the central metabolism were calculated from growth kinetics and analyzed in relation to specific enzyme activities. at high growth rates, incomplete glucose removal was observed, and this was attributed to rate-limiting capacity of the phosphotransferase system transporter and the probable contribution of a low-aff ...	1996	16535231
mechanism and regulation of isoleucine excretion in corynebacterium glutamicum.	whole cells of corynebacterium glutamicum were loaded with high cytoplasmic l-isoleucine concentrations, and isoleucine excretion from these cells was studied in terms of mechanism and regulation. the transmembrane isoleucine flux could be differentiated into carrier-mediated uptake, carrier-mediated excretion, and diffusion. after discrimination from the other transmembrane solute movements, the outward-directed flux, which was due to the activity of the isoleucine excretion carrier, was charac ...	1996	16535397
complete sucrose metabolism requires fructose phosphotransferase activity in corynebacterium glutamicum to ensure phosphorylation of liberated fructose.	sucrose uptake by corynebacterium glutamicum involves a phosphoenolpyruvate-dependent sucrose phosphotransferase (pts), but in the absence of fructokinase, further metabolism of the liberated fructose requires efflux of the fructose and reassimilation via the fructose pts. mutant strains lacking detectable fructose-transporting pts activity accumulated fructose extracellularly but consumed sucrose at rates comparable to those of the wild-type strain.	1996	16535429
determination of the fluxes in the central metabolism of corynebacterium glutamicum by nuclear magnetic resonance spectroscopy combined with metabolite balancing.	to determine the in vivo fluxes of the central metabolism we have developed a comprehensive approach exclusively based on the fundamental enzyme reactions known to be present, the fate of the carbon atoms of individual reactions, and the metabolite balance of the culture. no information on the energy balance is required, nor information on enzyme activities, or the directionalities of reactions. our approach combines the power of (1)h-detected (13)c nuclear magnetic resonance spectroscopy to fol ...	1996	18623562
development and application of a membrane cyclone reactor for in vivo nmr spectroscopy with high microbial cell densities.	a new bioreactor system has been developed for in vivo nmr spectroscopy of microorganisms under defined physiological conditions. this cyclone reactor with an integrated nmr flow cell is continuously operated in the magnet of a 400-mhz wide-bore nmr spectrometer system. the residence times of medium and cells are decoupled by a circulation-integrated cross-flow microfiltration module to achieve higher cell densities as compared to continuous fermentations without cell retention (increase in cell ...	1996	18629829
water reuse in the l-lysine fermentation process.	l-lysine is produced commercially by fermentation. as is typical for fermentation processes, a large amount of liquid waste is generated. to minimize the waste, which is mostly the broth effluent from the cation exchange column used for l-lysine recovery, we investigated a strategy of recycling a large fraction of this broth effluent to the subsequent fermentation. this was done on a labscale process with corynebacterium glutamicum atcc 21253 as the l-lysine-producing organism. broth effluent fr ...	1996	18623586
reaction engineering analysis of l-lysine transport by corynebacterium glutamicum.	to identify potential l-lysine export limitations by corynebacterium glutamicum in the l-lysine production process, the excretion of l-lysine was studied in continuous and fed-batch operated stirred tank reactors. a structured biochemical model of the l-lysine excretion mechanism was used to determine the activity of the export carrier and to calculate a cell-specific concentration of the export carrier. for the biochemical characterization of this specific carrier concentration a standardized l ...	1996	18627086
cloning and characterization of an is-like element present in the genome of brevibacterium lactofermentum atcc 13869.	a repetitive dna element of the gram+ brevibacterium lactofermentum (bl), cloned by a modification of the subtractive hybridization method, contained a 1.4-kb is-like element, is13869, which included an open reading frame (orf) inside a perfect 26-bp terminal inverted repeat (tir). an 8-bp direct repeat (dr) was found outside each tir. the orf encoded a deduced protein of 436 amino acids (49 380 da) with extensive similarity to other known transposases of insertion elements of mycobacterium smeg ...	1996	8621097
promoters from corynebacterium glutamicum: cloning, molecular analysis and search for a consensus motif.	relatively limited information about promoter structures in corynebacterium glutamicum has been available until now. with the aim of isolating and characterizing such transcription initiation signals, random sau3a fragments of c. glutamicum chromosomal dna and of the corynebacterial phage phi ga1 were cloned into the promoter probe vector pekplcm and selected for promoter activity by chloramphenicol resistance of transformed c. glutamicum cells. the nucleotide sequence of ten chromosomal and thr ...	1996	8704969
a physical and genetic map of the corynebacterium glutamicum atcc 13032 chromosome.	a combined physical and genetic map of the corynebacterium glutamicum atcc 13032 chromosome was constructed using pulsed-field gel electrophoresis (pfge) and hybridizations with cloned gene probes. total genomic dna was digested with the meganucleases swai (5'-atttaaat-3'), paci (5'-ttaattaa-3'), and pmei (5'-gtttaaac-3') yielding 26,27, and 23 fragments, respectively. the chromosomal restriction fragments were then separated by pfge. by summing up the lengths of the fragments generated with eac ...	1996	8842145
expression, purification, and crystallization of meso-diaminopimelate dehydrogenase from corynebacterium glutamicum.	the gene encoding the meso-diaminopimelate dehydrogenase (dapdh) from corynebacterium glutamicum was over-expressed and purified to homogeneity. crystals of the binary dapdh-nadp+ complex were obtained from solutions of polyethylene glycol 8000, 100 mm sodium cacodylate, ph 6.5, and 150-300 mm mg(oac)2. the crystals diffract to 2.2 a, belong to the orthorhombic space group p2(1), and contain two molecules per asymmetric unit.	1996	8865347
construction of l-lysine-, l-threonine-, and l-isoleucine-overproducing strains of corynebacterium glutamicum.	the gram-negative bacterium corynebacterium glutamicum is used for the industrial production of amino acids, for example, of l-glutamate and l-lysine. by cloning and expressing the various genes of the l-lysine pathway in c. glutamicum, we would demonstrate that an increase of the flux of l-aspartate semialdehyde to l-lysine could be obtained in strains with increased dihydrodipicolinate synthase activity. recently we detected that in c. glutamicum two pathways exist for synthesis of d,l-diamino ...	1996	8659901
functional and genetic characterization of the (methyl)ammonium uptake carrier of corynebacterium glutamicum.	under nitrogen starvation conditions, corynebacterium glutamicum was found to take up methylammonium at a rate of 20 +/- 5 nmol.min-1.(mg dry weight)-1. the specific activity of this uptake was 10-fold lower when growing the cells under sufficient nitrogen supply, indicating a tight regulation on the expression level. the methylammonium uptake showed michaelis-menten kinetics with an km of 44 +/- 7 microm and was completely inhibited by the addition of 10 microm ammonium. this finding and the fa ...	1996	8621394
mutations in the corynebacterium glutamicum proline biosynthetic pathway: a natural bypass of th proa step.	two chromosomal loci containing the corynebacterium glutamicum atcc 17965 prob and proc genes were isolated by complementation of escherichia coli prob and proc auxotrophic mutants. together with a proa gene described earlier, these new genes describe the major c. glutamicum proline biosynthetic pathway. the prob and proa genes, closely linked in most bacteria, are in c. glutamicum separated by a 304-amino-acid open reading frame (unk) whose predicted sequence resembles that of the 2-hydroxy aci ...	1996	8755867
characterization of the plasmid pmb1 from bifidobacterium longum and its use for shuttle vector construction.	the nucleotide sequence of the 1847-bp bifidobacterium longum b2577 cryptic plasmid pmb1 was determined. the plasmid had a g+c content of 62.0%, and contained two open reading frames, orf1 and orf2, likely arranged in an operon. the proteins encoded by orf1 and orf2 show the highest degree of similarity with similarly arranged peptide sequences translated from corynebacterium glutamicum pxz10142 and mycobacterium fortuitum pal5000 plasmids. recombinant plasmids containing the pmb1 replicon were ...	1996	8761732
construction of l-lysine-overproducing strains of brevibacterium lactofermentum by targeted disruption of the hom and thrb genes.	the mobilization of plasmids from gram-negative escherichia coli to gram-positive brevibacterium lactofermentum, mediated by p-type transfer functions, was used to construct disrupted mutants blocked specifically in the homoserine branch of the aspartate pathway. the mutant strain b. lactofermentum r31 showed an efficiency of conjugal transfer two to three orders of magnitude higher than that of the wild-type strain b. lactofermentum atcc 13869. the hom- and thrb-disrupted mutants of b. lactofer ...	1996	9008889
analysis of nucleotide methylation in dna from corynebacterium glutamicum and related species.	plasmid dna (pcsl17) isolated from corynebacterium glutamicum transformed recipient mcrbc+ strains of escherichia coli with lower efficiency than mcrbc- strains, confirming a previous report by tauch et al. (fems microbiol. lett. 123 (1994) 343-348) which inferred that c. glutamicum dna contains methylcytidine. analysis of nucleotides in c. glutamicum-derived chromosomal and plasmid dna failed to detect significant levels of methylated adenosine, but methylated cytidine was readily detected. res ...	1996	8867385
three-dimensional structure of meso-diaminopimelic acid dehydrogenase from corynebacterium glutamicum.	diaminopimelate dehydrogenase catalyzes the nadph-dependent reduction of ammonia and l-2-amino-6-ketopimelate to form meso-diaminopimelate, the direct precursor of l-lysine in the bacterial lysine biosynthetic pathway. since mammals lack this metabolic pathway inhibitors of enzymes in this pathway may be useful as antibiotics or herbicides. diaminopimelate dehydrogenase catalyzes the only oxidative deamination of an amino acid of d configuration and must additionally distinguish between two chir ...	1996	8885833
cloning of the tryptophan operon of brevibacterium divaricatum and its expression in e. coli.	a genomic bank from brevibacterium divaricatum has been prepared using lambda embl3 as a vector. the genomic bank's titers are 2.2 x 10(6) pfu/micrograms. through screening by plaque hybridization, a 9.6 kb ncoi fragment which contains the entire trp operon has been isolated. polymerase chain reaction amplification and restriction endonuclease analysis of pcr fragments indicated that there is homology between the coryneform bacteria; however, some genetic diversity among the species still exists ...	1996	8956524
a new type of transporter with a new type of cellular function: l-lysine export from corynebacterium glutamicum.	we discovered that after deregulation of the l-lysine biosynthesis in corynebacterium glutamicum, l-lysine accumulated in the cytosol and the efflux properties of this amino acid in mutants used for l-lysine production were altered. in this study we describe the cloning and molecular identification of lyse, which encodes the translocator specifically exporting l-lysine from the cell. the lyse gene product does not display homology to any known transporter. it is only 236 amino acids in size, wit ...	1996	8971704
c3-carboxylation as an anaplerotic reaction in phosphoenolpyruvate carboxylase-deficient corynebacterium glutamicum.	phosphoenolpyruvate carboxylase (pepcx) has recently been found to be dispensable as an anaplerotic enzyme for growth and lysine production of corynebacterium glutamicum. to clarify the role of the glyoxylate cycle as a possible alternative anaplerotic sequence, defined pepcx- and isocitrate-lyase (icl)-negative double mutants of c. glutamicum wild-type and of the l-lysine-producing strain mh20-22b were constructed by disruption of the respective genes. analysis of these mutants revealed that th ...	1996	8661932
genetic characterization of site-specific integration functions of phi aau2 infecting "arthrobacter aureus" c70.	all the essential genetic determinants for site-specific integration of corynephage phi aau2 are contained within a 1,756-bp dna fragment, carried on the integrative plasmid p5510, and are shown to be functional in escherichia coli. one open reading frame, orf4, encoding a protein of 266 amino acids was shown to represent the phi aau2 integrase. the nucleotide sequence of the phi aau2 attachment site, attp, and the attb, attl, and attr sequences in the host "arthrobacter aureus" c70 were determi ...	1996	8606175
transport mutants and transport genes of corynebacterium glutamicum.		1996	8659896
isolation, characterization, and expression of the corynebacterium glutamicum betp gene, encoding the transport system for the compatible solute glycine betaine.	corynebacterium glutamicum accumulates glycine betaine under conditions of high osmolarity. previous work revealed the existence of a high-affinity glycine betaine permease which is osmotically regulated. in the present study, the corresponding gene was cloned. the betp gene, encoding the glycine betaine uptake carrier, was isolated by heterologous complementation of mutant strain escherichia coli mkh13. from sequence analysis it is predicted to encode a protein of 595 amino acids. this protein ...	1996	8752342
construction of new cloning vectors for brevibacterium lactofermentum.	two plasmid cloning vectors (pulmj55 and pulmj95) were constructed for brevibacterium lactofermentum using the origin of replication of the endogenous plasmid pbl1. plasmid pulmj55 is a replacement vector with transcriptional terminators from the b. lactofermentum trp operon flanking the bglii cloning sites. religation of the bglii digested vector without insert creates a 376 bp perfect palindrome that is not tolerated in b. lactofermentum, giving positive selection for recombinant plasmids with ...	1996	8935658
codon usage in the mycobacterium tuberculosis complex.	the usage of alternative synonymous codons in mycobacterium tuberculosis (and m. bovis) genes has been investigated. this species is a member of the high-g+c gram-positive bacteria, with a genomic g+c content around 65 mol%. this g+c-richness is reflected in a strong bias towards c- and g-ending codons for every amino acid: overall, the g+c content at the third positions of codons is 83%. however, there is significant variation in codon usage patterns among genes, which appears to be associated ...	1996	8936318
multiple sigma factor genes in brevibacterium lactofermentum: characterization of siga and sigb.	four rpod hybridizing signals have been identified in the chromosome of brevibacterium lactofermentum. two rpod-like genes, siga and sigb, have been cloned and sequenced, and they encode principal sigma factors of the rna polymerase. the deduced amino acid sequences of siga and sigb showed very high similarities to those of mycobacterium smegmatis mysa and mysb proteins, respectively, and also to those of hrdb proteins from different streptomyces species. siga and sigb maintain the conserved mot ...	1996	8550480
genes and enzymes of the acetyl cycle of arginine biosynthesis in corynebacterium glutamicum: enzyme evolution in the early steps of the arginine pathway.	a cluster of arginine biosynthetic genes of corynebacterium glutamicum atcc 13032, comprising argj, argb and argd as well as part of argc and argf, has been cloned by heterologous complementation of an escherichia coli arge mutant. the gene order has been established as argcjbdf by sequencing the entire 4.4 kb cloned dna fragment. the c. glutamicum argb gene can be transcribed in e. coli cells from an internal promoter located in the coding part of the preceding argj gene, whereas transcription ...	1996	8581175
cloning of m-fluorophenylalanine-resistant gene and mutational analysis of feedback-resistant prephenate dehydratase from corynebacterium glutamicum.	corynebacterium glutamicum was mutated by nitrosoguanidine and five m-fluorophenylalanine (mfp)-resistant mutants were isolated. the mutants were resistant to phenylalanine-mediated feedback inhibition of the prephenate dehydratase activity. cloning and characterization of the mfp-resistant gene revealed that mutant prephenate dehydratase, encoded by the phe a gene, confers the mfp-resistant phenotype upon c. glutamicum. to determine the amino acid residues to which variation may result in the f ...	1996	8605023
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 ...	1996	8917070
molecular cloning of a novel gene, dtsr, which rescues the detergent sensitivity of a mutant derived from brevibacterium lactofermentum.	several strains of corynebacterium and brevibacterium are known for their ability to secrete large amounts of amino acids, especially l-glutamate. we focused on the mechanism of l-glutamate secretion triggered by a detergent, namely polyoxyethylenesorbitan monopalmitate (pesp). a mutant strain, aj11060, derived from brevibacterium lactofermentum atcc 13869 indicates the sensitivity to pesp. a multicopy suppresser gene that compliments the sensitivity of aj11060 to the detergent was derived from ...	1996	8987652
molecular cloning of the corynebacterium glutamicum ('brevibacterium lactofermentum' aj12036) odha gene encoding a novel type of 2-oxoglutarate dehydrogenase.	the corynebacterium glutamicum ('brevibacterium lactofermentum' aj12036) odha gene, encoding 2-oxoglutarate dehydrogenase (e1o subunit of the 2-oxoglutarate dehydrogenase complex), has been isolated and identified as an homologous counterpart of the escherichia coll suca and bacillus subtilis odha genes. the nucleotide sequence of a 4394 bp chromosomal fragment containing the c. glutamicum odha gene was determined. the odha gene comprised 3771 bp (1257 codons, including the initiation codon) and ...	1996	9004499
the gale gene encoding the udp-galactose 4-epimerase of brevibacterium lactofermentum is coupled transcriptionally to the dmdr gene.	the gale gene of brevibacterium lactofermentum, encoding udp-galactose 4-epimerase (ec 5.1.3.2), has been identified by dna sequencing downstream from the orf1-sigb-dmdr region. the arrangement of the sigb-dtxr-gale cluster is also conserved in corynebacterium diphtheriae. the deduced gale product was a protein of 329 aa residues (35.4 kda) that shared a high degree of identity to known udp-galactose 4-epimerase proteins from gram-positive microorganisms (streptomyces lividans and streptococcus ...	1996	8921853
a brevibacterium linens prbl1 replicon functional in corynebacterium glutamicum.	brevibacterium linens rbl strain cryptic plasmid prbl1 (8.0 kb) is described. a region involved in prbl1 autonomous replication in corynebacterium glutamicum was identified by insertion and deletion mapping and partially sequenced. this region encodes for a hypothetical 310-amino acid (aa) protein closely related to the replicases of plasmids pxz10142 (c. glutamicum) and pal5000 (mycobacterium fortuitum). the 310-aa protein also shows significant homology to proteins of pcole5-099 (shigella sonn ...	1996	8938050
effects of altered phosphoenolpyruvate carboxylase activities on transgenic c3 plant solanum tuberosum.	phosphoenolpyruvate carboxylase (pepc) genes from corynebacterium glutamicum (cppc), escherichia coli (eppc) or flaveria trinervia (fppc) were transferred to solanum tuberosum. plant regenerants producing foreign pepc were identified by western blot analysis. maximum pepc activities measured in eppc and fppc plants grown in the greenhouse were doubled compared to control plants. for cppc a transgenic plant line could be selected which exhibited a fourfold increase in pepc activity. in the presen ...	1996	8980535
a corynebacterium glutamicum gene encoding a two-domain protein similar to biotin carboxylases and biotin-carboxyl-carrier proteins.	following the analysis of transposon tn5432-induced mutants of corynebacterium glutamicum atcc 13032, a gene encoding a protein with a biotin-binding motif was cloned. the dna sequence of this gene revealed an open reading frame encoding 591 amino acids with a calculated mol. mass of 63.4 kda. the protein is composed of two domains, an n-terminal biotin carboxylase and a c-terminal biotin-carboxyl-carrier protein, that are highly similar to corresponding subunits from prokaryotic and eukaryotic ...	1996	8772169
a cryptic plasmid pbl1 from brevibacterium lactofermentum causes growth inhibition and filamentation in escherichia coli.	a shuttle vector composed of pbl1, a 4.46-kb cryptic plasmid from coryneform bacterium brevibacterium lactofermentum, and escherichia coli vector phsg298 was found to inhibit growth and cause cell filamentation in e coli. after construction of several deletions of pbl1, a 1.23-kb acci-hindiii fragment was found responsible. dna sequence analysis showed that this fragment contained a 726-bp-long open reading frame (orf3), encoding a protein with 242 amino acid residues. corresponding to orf3, a 2 ...	1996	8938054
a corynebacterium glutamicum gene conferring multidrug resistance in the heterologous host escherichia coli.	a chromosomal dna fragment from the erythromycin-sensitive bacterium corynebacterium glutamicum atcc 13032 was shown to mediate resistance against erythromycin, tetracycline, puromycin, and bleomycin in escherichia coli. multicopy cloning of the fragment did not cause a resistance phenotype in c. glutamicum. the corresponding gene encodes a hydrophobic protein with 12 potential transmembrane-spanning ex-helical segments showing similarity to drug-h+ antiporters.	1997	9079937
plasmid pga1 from corynebacterium glutamicum codes for a gene product that positively influences plasmid copy number.	the complete nucleotide sequence (4,826 bp) of the cryptic plasmid pga1 from corynebacterium glutamicum was determined. dna sequence analysis revealed four putative coding regions (open reading frame a [orfa], orfa2, orfb, and orfc). orfc was identified as a rep gene coding for an initiator of plasmid replication (rep) according to the high level of homology of its deduced amino acid sequence with the rep proteins of plasmids psr1 (from c. glutamicum) and png2 (from corynebacterium diphtheriae). ...	1997	9045809
accurate determination of 13c enrichments in nonprotonated carbon atoms of isotopically enriched amino acids by 1h nuclear magnetic resonance.	a method for the accurate determination of 13c enrichments in nonprotonated carbon atoms of organic compounds that makes use of unresolved 13c satellites of proton(s) bonded to the vicinal carbon atom was developed. using glutamate as a model molecule, this 1h nuclear magnetic resonance (nmr) inverse spin-echo difference spectroscopy method was calibrated for inversion efficiency and relaxation effects which were then shown to cause only a minor loss of the measured 13c satellite amplitude (2% f ...	1997	9056211
[the serological properties of saprophytic corynebacteria studied by immunoenzyme analysis].	the degree of serological similarity of saprophytic corynebacteria have been studied using immunoassay elisa analysis, that is seven collection strains, belonging to corynebacterium glutamicum (3 strains), c. ammoniagenes (1 strain), c. vitarumen (1 strain), c. variabilis (2 strains) and three industrial strains-lysine producers. intact and heated bacteria cells have been used as antigens. it has been shown that industrial strain c. glutamicum 22l and collection strains c. glutamicum imv ac-715, ...	1997	9480013
a dtsr gene-disrupted mutant of brevibacterium lactofermentum requires fatty acids for growth and efficiently produces l-glutamate in the presence of an excess of biotin.	a dtsr gene encoding a homolog of the beta subunit of some biotin-containing enzymes suppresses a detergent-sensitive mutation of brevibacterium lactofermentum (e. kimura et al., 1996, biosci. biotech. biochem. 60, 1565-1570), which has been used for the fermentative production of l-glutamate. when the dtsr gene was disrupted, the organism exhibited strict fatty acid auxotrophy; oleate or oleate ester, but not palmitate ester or stearate ester, supported the growth of the delta dtsr mutant. immu ...	1997	9168981
efflux of compatible solutes in corynebacterium glutamicum mediated by osmoregulated channel activity.	bacteria respond to hypoosmotic stress by releasing low-molecular-mass solutes in order to maintain constant turgor pressure. we have studied the function of osmoregulated channel(s) in corynebacterium glutamicum, which are responsible for efflux of various solutes upon sudden decrease in osmotic pressure. the channels preferentially mediated efflux of compatible solutes such as glycine betaine and proline. the release of molecules of similar size, e.g. glutamate or lysine, was restricted, atp w ...	1997	9266699
isolation of the putp gene of corynebacterium glutamicum and characterization of a low-affinity uptake system for compatible solutes.	corynebacterium glutamicum accumulates the compatible solutes proline, glycine betaine, and ectoine under conditions of high osmolality. uptake of proline is mediated by both a high-affinity and a low-affinity secondary transport system. the low-affinity uptake system also accepts glycine betaine and ectoine as substrates. in the present study, the gene encoding the high-affinity proline uptake system putp was isolated by heterologous complementation of escherichia coli mutant strain wg389, whic ...	1997	9238106
relationship between the glutamate production and the activity of 2-oxoglutarate dehydrogenase in brevibacterium lactofermentum.	enzyme activities of 2-oxoglutarate dehydrogenase complex and glutamate dehydrogenase of wild type brevibacterium lactofermentum, one of the typical glutamate-producing coryneform bacteria, were investigated by using cells cultured under glutamate-productive and glutamate-non-productive conditions. significant reduction of the former enzyme activity was observed in the cells under the several glutamate-productive conditions, namely, in the cells cultured in media containing a) limited concentrat ...	1997	9255973
the s-layer protein of corynebacterium glutamicum is anchored to the cell wall by its c-terminal hydrophobic domain.	ps2 is the s-layer protein of corynebacterium glutamicum. the s-layer may be detached from the cell as organized sheets by detergents at room temperature. the solubilization of ps2 in the form of monomers requires detergent treatment at high temperature (70 degrees c), conditions under which the protein is denatured. treatment of the cells with proteinase k or trypsin results in the detachment of the organized s-layer, which remains organized. because we show that trypsin cleaves the c-terminal ...	1997	9044282
isolation of the corynebacterium glutamicum glna gene encoding glutamine synthetase i.	the corynebacterium glutamicum glutamine synthetase i (gsi) structural gene glna was cloned by a pcr approach using oligonucleotide primers derived from conserved amino acid sequences of the gsi proteins from various bacteria. disruption or deletion of this gene in c. glutamicum led to a glutamine auxotrophic phenotype and complete loss of glutamine synthetase activity, indicating the key role of this enzyme in nitrogen metabolism. additionally, indications for a second glutamine synthetase, gsi ...	1997	9297824
sequence analysis of functional regions of homoserine dehydrogenase genes from l-lysine and l-threonine-producing mutants of brevibacterium lactofermentum.	the homoserine dehydrogenase (hd) genes from brevibacterium lactofermentum lysine- and threonine-producing mutants were cloned, using the polymerase chain reaction, and sequenced. we found the amino acid substitutions, val104ile in the lysine-producing mutants in which hd may cause leaky mutation and ser393phe in the threonine-producing mutant with feedback-insensitive hd.	1997	9362124
the corynebacterium glutamicum cglim gene encoding a 5-cytosine methyltransferase enzyme confers a specific dna methylation pattern in an mcrbc-deficient escherichia coli strain.	the cglim gene of the coryneform soil bacterium corynebacterium glutamicum atcc 13032 has been cloned and characterized. the coding region comprises 1092 nucleotides and specifies a protein of 363 amino acid residues with a deduced mr of 40700. the amino acid sequence showed striking similarities to methyltransferase enzymes generating 5-methylcytosine residues, especially to m x ngovii from neisseria gonorrhoeae recognizing the sequence gcsgc. the cglim gene is organized in an unusual operon wh ...	1997	9426239
the monovalent cation requirement of rabbit muscle pyruvate kinase is eliminated by substitution of lysine for glutamate 117.	the crystal structure of rabbit muscle pyruvate kinase complexed with mn2+, k+, and pyruvate revealed a binding site of k+ [t. m. larsen, l. t. laughlin, h. m. holden, i. rayment, and g. h. reed (1994) biochemistry 33, 6301-6309]. sequence comparisons of rabbit muscle pyruvate kinase and pyruvate kinases from corynebacterium glutamicum and escherichia coli, which do not exhibit a requirement for activation by monovalent cations, indicate that the only substitutions in the k+ binding site are con ...	1997	9434737
ultrastructure of the corynebacterium glutamicum cell wall.	the cell wall structure of the gram-positive corynebacterium glutamicum was evaluated by electron microscopy of thin sections after freeze-substitution and conventional fixation with glutaraldehyde. for the cell wall an overall thickness of approximately 32 nm was determined, with 8.5 nm corresponding to an outer layer, 6.5 nm to an electron translucent region (etr) as found in mycobacteria and 17 nm to the peptidoglycan. knob-like surface structures previously observed in freeze-fracture experi ...	1997	9442270
drug extrusion in corynebacterium glutamicum.	we selected a mutant of corynebacterium glutamicum, ebr, which can grow in a medium containing cytotoxic ethidium bromide (etbr) at a high concentration of 100 microm. the resistance to etbr in the mutant was reversed by 2 microm reserpine, a potent inhibitor of mammalian p-glycoprotein and bacterial multidrug resistance (mdr) transporter, whereas reserpine alone had a minimal effect on cell growth. the mutant showed a much higher efflux rate of etbr than wild-type cells, and the efflux was comp ...	1997	9442486
transcriptional analysis of the siga and sigb genes of brevibacterium lactofermentum.	transcription of the siga and sigb genes of brevibacterium lactofermentum, encoding the principal sigma factors of rna polymerase, has been investigated by northern blot and primer extension analysis. northern hybridizations revealed that siga is transcribed as a monocistronic mrna of 1.7 kb and sigb gives two transcripts of 1.1 and 1.5 kb. similar transcription patterns of siga and sigb genes in nutrient-rich medium were observed; transcripts of both genes occurred simultaneously throughout the ...	1997	9252580
regulation of acetate metabolism in corynebacterium glutamicum: transcriptional control of the isocitrate lyase and malate synthase genes.	in the amino-acid-producing microorganism corynebacterium glutamicum, the specific activities of the acetate-activating enzymes acetate kinase and phosphotransacetylase and those of the glyoxylate cycle enzymes isocitrate lyase and malate synthase were found to be high when the cells were grown on acetate (0.8, 2.9, 2.1, and 1.8 u/mg protein, respectively). when the cells were grown on glucose or on other carbon sources such as lactate, succinate, or glutamate, the specific activities were two- ...	1997	9297462
heterologous expression of the mycobacterium tuberculosis gene encoding antigen 85a in corynebacterium glutamicum.	by using appropriate corynebacterium glutamicum-escherichia coli shuttle plasmids, the gene encoding the fibronectin-binding protein 85a (85a) from mycobacterium tuberculosis was expressed in c. glutamicum, also an actinomycete and nonsporulating gram-positive rod bacterium, which is widely used in industrial amino acid production. the 85a gene was weakly expressed in c. glutamicum under the control of the ptac promoter from e. coli, but it was produced efficiently under the control of the promo ...	1997	9361426
molecular biology of s-layers.	in this chapter we report on the molecular biology of crystalline surface layers of different bacterial groups. the limited information indicates that there are many variations on a common theme. sequence variety, antigenic diversity, gene expression, rearrangements, influence of environmental factors and applied aspects are addressed. there is considerable variety in the s-layer composition, which was elucidated by sequence analysis of the corresponding genes. in corynebacterium glutamicum one ...	1997	9276928
cloning, sequencing and expression of the gene encoding elongation factor p in the amino-acid producer brevibacterium lactofermentum (corynebacterium glutamicum atcc 13869).	the brevibacterium lactofermentum ef-p gene, encoding the elongation factor protein p, was cloned and sequenced. according to dna sequence analysis of this gene, the b. lactofermentum ef-p protein consists of 187 amino acids with a calculated molecular weight of 20,584. southern hybridization of an internal fragment of the ef-p gene from b. lactofermentum with chromosomal dnas from different microorganisms reveals that it is a unique gene product in b. lactofermentum and corynebacterium glutamic ...	1997	9370284
an on-line physiological state recognition system for the lysine fermentation process based on a metabolic reaction model.	a metabolic reaction model was developed for the lysine fermentation process by corynebacterium glutamicum aj-3462 to estimate the physiological state of the cells-that is, the growth and production activity, and the flux distribution of metabolites-from on-line measurable rates only. first, the extended kalman filter was applied to eliminate noise in the measured rates. then, using the metabolic reaction model, the lysine production rate and flux distribution were calculated. the estimation res ...	1997	18636455
bidirectional reaction steps in metabolic networks: ii. flux estimation and statistical analysis.	metabolic carbon labelling experiments enable a large amount of extracellular fluxes and intracellular carbon isotope enrichments to be measured. since the relation between the measured quantities and the unknown intracellular metabolic fluxes is given by bilinear balance equations, flux determination from this data set requires the numerical solution of a nonlinear inverse problem. to this end, a general algorithm for flux estimation from metabolic carbon labelling experiments based on the leas ...	1997	18636450
response of the central metabolism of corynebacterium glutamicum to different flux burdens.	to evaluate the importance of reactions within the central metabolism under different flux burdens the fluxes within the pentose phosphate pathway (ppp), as well as the other reactions of the central metabolism, were intensively analyzed and quantitated. for this purpose, corynebacterium glutamicum was grown with [1-(13)c]glucose to metabolic and isotopic steady state and the fractional enrichments in precursor metabolites (e.g., pentose 5-phosphate) were quantified. matrix calculus was used to ...	1997	18636622
metabolic and physiological studies of corynebacterium glutamicum mutants.	the physiology and central carbon metabolism of corynebacterium glutamicum was investigated through the study of specific disruption mutants. mutants deficient in phosphoenolpyruvate carboxylase (ppc) and/or pyruvate kinase (pk) activity were constructed by disrupting the corresponding gene(s) via transconjugation. standard batch fermentations were carried out with these mutants and results were evaluated in the context of intracellular flux analysis. the following were determined. (a) there is ...	1997	18636597
in situ global method for measurement of oxygen demand and mass transfer.	two aerobic microorganisms, saccharomycopsis lipolytica and brevibacterium lactofermentum, have been used in a study of mass transfer and oxygen uptake from a global perspective, using a closed gas system. oxygen concentrations in the gas and liquid were followed using oxygen electrodes; the results allowed for easy calculation of in situ oxygen transport. the cell yields on oxygen for s. lipolytica and b. lactofermentum were 1.01 and 1.53 g/g, respectively. the mass transfer coefficient was est ...	1998	18576018
osmo-sensing by n- and c-terminal extensions of the glycine betaine uptake system betp of corynebacterium glutamicum.	the major uptake carrier for the compatible solute glycine betaine in corynebacterium glutamicum is the secondary transport system betp. it is effectively regulated by the external osmolality both on the level of expression and of activity. betp carries highly charged domains both at the n and at the c terminus. we investigated the role of these extensions in the regulatory response to hyperosmotic stress. mutants of the betp gene coding for proteins with truncated n- and c-terminal extensions w ...	1998	9446558
improved l-lysine yield with corynebacterium glutamicum: use of dapa resulting in increased flux combined with growth limitation.	the amino acid l-lysine is produced on a large scale using mutants of corynebacterium glutamicum. however, as yet recombinant dna techniques have not succeed in improving strains selected for decades by classic mutagenesis for high productivity. we here report that seven biosynthetic enzymes were assayed and oversynthesis of the dihydrodipicolinate synthase resulted in an increase of lysine accumulation from 220 mm to 270 mm. the synthase, encoded by dapa, is located at the branch point of metab ...	1998	9487706
substrate and inhibitor binding sites in corynebacterium glutamicum diaminopimelate dehydrogenase.	the three-dimensional structures of corynebacterium glutamicum diaminopimelate dehydrogenase as a binary complex with the substrate meso-diaminopimelate (meso-dap) and a ternary complex with nadp+ and an isoxazoline inhibitor [abbot, s.d., lane-bell, p., kanwar, p.s.s., and vederas, j. c. (1994) j. am. chem. soc. 116, 6513-6520] have been solved and refined against x-ray diffraction data to 2.2 a. diaminopimelate dehydrogenase is a homodimer of approximately 35,000 molecular weight subunits and ...	1998	9521647
a bacterial cytokine.	viable cells of micrococcus luteus secrete a factor, which promotes the resuscitation and growth of dormant, nongrowing cells of the same organism. the resuscitation-promoting factor (rpf) is a protein, which has been purified to homogeneity. in picomolar concentrations, it increases the viable cell count of dormant m. luteus cultures at least 100-fold and can also stimulate the growth of viable cells. rpf also stimulates the growth of several other high g+c gram-positive organisms, including my ...	1998	9671779
isoleucine uptake in corynebacterium glutamicum atcc 13032 is directed by the brnq gene product.	by complementation analysis of an isoleucine-uptake-deficient escherichia coli strain, it was shown that a 1.6-kb hindiii-stui fragment of corynebacterium glutamicum atcc 13032, located downstream of the aecd gene, encodes an isoleucine uptake system. sequence analysis revealed that the complementing fragment carried an open reading frame, termed brnq, that encodes a protein with sequence similarities to branched-chain amino acid carriers of gram-positive and gram-negative bacteria. the brnq gen ...	1998	9531631
effect of reversible reactions on isotope label redistribution--analysis of the pentose phosphate pathway.	the pentose phosphate pathway plays several key roles in metabolism including supply of biosynthetic carbon skeletons and reducing power. previous research has focused on determining the fluxes through the reactions of this pathway using carbon-labeled substrates and models that make certain assumptions about the reversibility of the transketolase and transaldolase reactions in the nonoxidative pathway. these assumptions, however, have resulted in inconsistencies between the predicted carbon lab ...	1998	9546650
[construction and characteristics of a cosmid library of genes of the bacterium cornyebacterium glutamicum atss13032].	a representative genomic library of the corynebacterium glutamicum atcc 13032 genes in a cosmid vector lorist6 was created. the cosmids contain inserts of bacterial dna obtained by partial digestion with the sau3a i restrictase. five hundred and thirty individual primary recombinant clones were transferred into the wells of microtiter plates, where they are now being preserved. the average size of the bacterial dna inserts determined via a sum of restriction fragment sizes of recombinant molecul ...	1998	9589870
ppc, the gene for phosphoenolpyruvate carboxylase from an extremely thermophilic bacterium, rhodothermus obamensis: cloning, sequencing and overexpression in escherichia coli.	the ppc gene, which encodes phosphoenolpyruvate carboxylase (pepc) of an extremely thermophilic bacterium, rhodothermus obamensis, was directly sequenced by the thermal asymmetric interlaced (tail) pcr method. an orf for a 937 amino acid polypeptide was found in the gene. the ppc gene had a high g+c content (66.2 mol%) and the third position of the codon exhibited strong preference for g or c usage (85.0 mol%). the calculated molecular mass was 107,848 da, which was consistent with the molecular ...	1998	9611816
different modes of diaminopimelate synthesis and their role in cell wall integrity: a study with corynebacterium glutamicum.	in eubacteria, there are three slightly different pathways for the synthesis of m-diaminopimelate (m-dap), which is one of the key linking units of peptidoglycan. surprisingly, for unknown reasons, some bacteria use two of these pathways together. an example is corynebacterium glutamicum, which uses both the succinylase and dehydrogenase pathways for m-dap synthesis. in this study, we clone dapd and prove by enzyme experiments that this gene encodes the succinylase (m(r) = 24082), initiating the ...	1998	9620966
new shuttle vectors for rhodococcus sp. r312 (formerly brevibacterium sp. r312), a nitrile hydratase producing strain.	two shuttle vectors named prc52 (10.7 kb) and prk52 (12.2 kb) carrying chloramphenicol (cm) and chloramphenicol plus kanamycin (km) resistance genes, respectively, were constructed by fusion of a cryptic plasmid pbl13869 (replicon pbl1, 5.8 kb) from brevibacterium lactofermentum atcc13869 with pbr328 e. coli plasmid. transformation of rhodococcus sp. r312 (formerly brevibacterium sp. r312) protoplasts was realised with an efficiency of 28 transformants per micrograms of dna.	1998	9637010
cloning of the histidine biosynthetic genes of corynebacterium glutamicum: organization and sequencing analysis of the hisa, impa, and hisf gene cluster.	the hisa and hisf genes of corynebacterium glutamicum were cloned by transforming histidine auxotrophic escherichia coli with the genomic dna library. they are two of the eight genes that participate in the histidine biosynthetic pathway. cloned dna fragments containing the genes can also complement hish and hisi auxotrophs of escherichia coli, suggesting that the four genes are clustered in the genome. we determined the nucleotide sequences of the minimal fragment containing the hisa and hisf g ...	1998	9647764
cloning and characterization of 3-deoxy-d-manno-oct-2-ulosonic acid (kdo) transferase genes (kdta) from acinetobacter baumannii and acinetobacter haemolyticus.	3-deoxy-d-manno-oct-2-ulosonic acid (kdo) transferases (kdta) are multifunctional glycosyltransferases with primary structures of low similarity. totally degenerated primers were deduced from two stretches of identical amino acids between known kdta sequences and used to amplify by pcr a kdta-specific fragment from acinetobacter baumannii atcc 15308 dna which was then applied as a probe for the cloning and sequencing of the complete kdo transferase gene. with conserved pcr primers for this struc ...	1998	9660198
the role of the corynebacterium glutamicum rel gene in (p)ppgpp metabolism.	to investigate the metabolism of (p)ppgpp in amino-acid-producing coryneform bacteria, a pcr-based strategy using degenerate consensus oligonucleotides was applied to isolate the rel gene of corynebacterium glutamicum atcc 13032. the gene consists of 2283 nucleotides and encodes a protein of 760 amino acids with a molecular mass of 84.4 kda. the amino acid sequence revealed extensive similarities to the related proteins rela and spot of escherichia coli, which are known to be involved in (p)ppgp ...	1998	9695918
a novel system with positive selection for the chromosomal integration of replicative plasmid dna in corynebacterium glutamicum.	a simple system has been developed for generating corynebacterium glutamicum strains containing stable replicative plasmids integrated into the chromosome via homologous recombination. the system is based upon extremely strong incompatibility between two plasmids, which cannot be co-maintained even under antibiotic selective pressure. integration of the resident plasmid that contained the trpd gene of c. glutamicum was achieved by introduction of a second plasmid and subsequent selection for the ...	1998	9695919
analysis of the leub gene from corynebacterium glutamicum.	the leub gene of corynebacterium glutamicum was found to be present on a 2.2-kb bamhi-saci chromosomal fragment which complemented the leub mutation of escherichia coli. the activity of 3-isopropylmalate dehydrogenase (ec 1.1.1.85), encoded by the leub gene, was significantly increased in c. glutamicum cells harbouring a plasmid containing the 2.2-kb fragment. the nucleotide sequence of the c. glutamicum leub coding region (an open reading frame, orf, of 1020 bp encoding a polypeptide of 340 ami ...	1998	9720199
pyruvate carboxylase from corynebacterium glutamicum: characterization, expression and inactivation of the pyc gene.	in addition to phosphoenolpyruvate carboxylase (pepcx), pyruvate carboxylase (pcx) has recently been found as an anaplerotic enzyme in the amino-acid-producing bacterium corynebacterium glutamicum. using oligonucleotides designed according to conserved regions of pcx amino acid sequences from other organisms, a 200 bp fragment central to the c. glutamicum pcx gene (pyc) was amplified from genomic dna by pcr. this fragment was then used to identify and to subclone the entire c. glutamicum pyc gen ...	1998	9579065
biochemical and genetic characterization of the membrane-associated malate dehydrogenase (acceptor) from corynebacterium glutamicum.	in addition to a cytoplasmic, nad-dependent malate dehydrogenase (ec 1.1.1.37), corynebacterium glutamicum possesses a highly active membrane-associated malate dehydrogenase (acceptor) (ec 1.1.99.16). this enzyme also takes part in the citric acid cycle. it oxidizes l-malate to oxaloacetate and donates electrons to ubiquinone-1 and other artificial acceptors or, via the electron transfer chain, to oxygen. nad is not an acceptor and the natural direct acceptor for the enzyme is most likely a quin ...	1998	9660197
sequence of the corynebacterium glutamicum pyruvate carboxylase gene.	pyruvate carboxylase is an important anaplerotic enzyme replenishing oxaloacetate consumed for biosynthesis during growth, or lysine and glutamic acid production in industrial fermentations. we used regions of homology from pyruvate carboxylase sequences of 12 different species (corresponding to the atp- and pyruvate-binding sites), to design polymerase chain reaction (pcr) primers for amplifying a fragment of the pyruvate carboxylase (pc) gene from c. glutamicum genomic dna. this 850-base-pair ...	1998	9802220
corynebacterium striatum chloramphenicol resistance transposon tn5564: genetic organization and transposition in corynebacterium glutamicum.	the clinical isolate corynebacterium striatum m82b (formerly corynebacterium xerosis m82b) carries the 50-kb r-plasmid ptp10 conferring resistance to the antibiotics chloramphenicol, erythromycin, kanamycin, and tetracycline. dna sequence analysis of the chloramphenicol resistance region revealed the presence of the 4155-bp transposable element tn5564. the ends of tn5564 are identical 22-bp inverted repeats flanked by a 6-bp target site duplication. the central region of tn5564 encodes the chlor ...	1998	9735314
urea uptake and urease activity in corynebacterium glutamicum.	when corynebacterium glutamicum is grown with a sufficient nitrogen supply, urea crosses the cytoplasmic membrane by passive diffusion. a permeability coefficient for urea diffusion of 9 x 10(-7) cm s-1 was determined. under conditions of nitrogen starvation, an energy-dependent urea uptake system was synthesized. carrier-mediated urea transport was catalyzed by a secondary transport system linked with proton motive force. with a km for urea of 9 microm, the affinity of this uptake system was mu ...	1998	9560422
identification, characterization, and chromosomal organization of the ftsz gene from brevibacterium lactofermentum.	the ftsz gene was cloned from the chromosomal dna of brevibacterium lactofermentum by the polymerase chain reaction (pcr) using two oligonucleotides designed from two conserved regions found in most of the previously cloned and sequenced ftsz genes from other microorganisms. ftsz is a single-copy gene in corynebacteria and is located downstream from ftsq and murc, indicating linkage between genes involved in peptidoglycan synthesis (mur genes) and genes involved in cell division (fts genes). the ...	1998	9738885
biochemical and biophysical characterization of the cell wall porin of corynebacterium glutamicum: the channel is formed by a low molecular mass polypeptide.	the cell wall of the gram-positive bacterium corynebacterium glutamicum contains a channel (porin) for the passage of hydrophilic solutes. the channel-forming protein was identified, by lipid bilayer experiments, in the cell envelope fractions isolated by sucrose-density centrifugations and in organic solvent of whole cells. it was purified to homogeneity by fast-protein liquid chromatography across a mono-q column. the pure protein had a rather low molecular mass of about 5 kda as judged by sds ...	1998	9790664
carbon-flux distribution in the central metabolic pathways of corynebacterium glutamicum during growth on fructose.	growth of corynebacterium glutamicum on fructose was significantly less than that obtained on glucose, despite similar rates of substrate uptake. this was in part due to the production of overflow metabolites (dihydroxyacetone and lactate) but also to the increased production of co2 during growth on fructose. these differences in carbon-metabolite accumulation are indicative of a different pattern of carbon-flux distribution through the central metabolic pathways. growth on glucose has been prev ...	1998	9652400
corynebacterium glutamicum is equipped with four secondary carriers for compatible solutes: identification, sequencing, and characterization of the proline/ectoine uptake system, prop, and the ectoine/proline/glycine betaine carrier, ectp.	gram-positive soil bacterium corynebacterium glutamicum uses the compatible solutes glycine betaine, proline, and ectoine for protection against hyperosmotic shock. osmoregulated glycine betaine carrier betp and proline permease putp have been previously characterized; we have identified and characterized two additional osmoregulated secondary transporters for compatible solutes in c. glutamicum, namely, the proline/ectoine carrier, prop, and the ectoine/glycine betaine/proline carrier, ectp. a ...	1998	9811661
molecular cloning and analysis of the argc gene from corynebacterium glutamicum.	the argc gene encoding n-acetylglutamate 5-semialdehyde dehydrogenase has been cloned from corynebacterium glutamicum by transforming escherichia coli arginine auxotroph with the genomic dna library. based on the restriction map of the cloned dna, we have subcloned and sequenced the minimal dna fragment complementing the e. coli argc mutant. the coding region of the cloned gene is 1041 nucleotides long with a predicted molecular mass of about 38 kda polypeptide. enzyme activity and size of the e ...	1998	9818083
experimental determination of group flux control coefficients in metabolic networks	grouping of reactions around key metabolite branch points can facilitate the study of metabolic control of complex metabolic networks. this top-down metabolic control analysis is exemplified through the introduction of group (flux, as well as concentration) control coefficients whose magnitudes provide a measure of the relative impact of each reaction group on the overall network flux, as well as on the overall network stability, following enzymatic amplification. in this article, we demonstrate ...	1998	10191384
an integron of class 1 is present on the plasmid pcg4 from gram-positive bacterium corynebacterium glutamicum.	the streptomycin/spectinomycin resistance determinant of the 29-kb plasmid pcg4 from corynebacterium glutamicum was found to be a part of a typical class 1 integron. the sequence analysis revealed that the integron (designated incg) identified in this gram-positive bacterium is almost identical to the integron inc present on the plasmid psa1700 from the gram-negative bacterium pseudomonas aeruginosa. differences in only two base pairs were found in the 3.8-kb sequence. one base substitution (g-- ...	1998	9868786
isolation and analysis of meta, a methionine biosynthetic gene encoding homoserine acetyltransferase in corynebacterium glutamicum.	the meta gene encoding homoserine acetyltransferase, the first enzyme of the methionine biosynthetic pathway, was isolated from a pmt1-based corynebacterium glutamicum gene library via complementation of an escherichia coli meta mutant. a dna-sequence analysis of the cloned dna is identified an open-reading frame of 1,137 bp which encodes a protein with the molecular weight of 41,380 comprising 379 amino acids. the putative protein product showed good amino acid-sequence homology to its counterp ...	1998	9666465