Publications

activity regulation of the betaine transporter betp of corynebacterium glutamicum in response to osmotic compensation.	as a response to hyperosmotic stress bacterial cells accumulate compatible solutes by synthesis or by uptake. beside the instant activation of uptake systems after an osmotic upshift, transport systems show also a second, equally important type of regulation. in order to adapt the pool size of compatible solutes in the cytoplasm to the actual extent of osmotic stress, cells down-regulate solute uptake when the initial osmotic stress is compensated. here we describe the role of the betaine transp ...	2004	15581860
dynamics of glutamate synthesis and excretion fluxes in batch and continuous cultures of temperature-triggered corynebacterium glutamicum.	corynebacterium glutamicum 2262 strain, when triggered for glutamate excretion, experiences a rapid decrease in growth rate and increase in glutamate efflux. in order to gain a better quantitative understanding of the factors controlling the metabolic transition, the fermentation dynamics was investigated for a temperature-sensitive strain cultivated in batch and glucose-limited continuous cultures. for non-excreting cells at 33 degrees c, increasing the growth rate resulted in strong increases ...	2004	15614534
a high-resolution titrator: a new approach to studying binding sites of microbial biosorbents.	the high-resolution potentiometric titration was used as a physico-chemical method to study the acid properties of selected biosorbent materials in order to quantify the functional acidic groups for sorption and to determine their affinities by considering their partial or total ionization equilibrium reactions. the gran's method and the henderson-hasselbach's equation were employed in establishing the partition of the total acidity as associated with strong, weak and very weak acidic chemical a ...	2004	15707630
purification of l-lysine in simulated moving bed and fixed-bed chromatography.	l-lysine was produced by a microbial process utilizing a corynebacterium glutamicum (atcc 21799) strain. l-lysine was purified from the cultivated medium by fixed-bed and simulated moving bed (smb) chromatography. the separation conditions including ph, eluent concentration and lys+ and lys2+ adsorption isotherms were studied in batch adsorption. the column capacity, eluent flow rate and eluent concentration have been studied in fixed-bed chromatography. maximum purification rate of lysine was o ...	2004	15709427
cre/loxp-mediated deletion system for large genome rearrangements in corynebacterium glutamicum.	genome rearrangement is an increasingly important technique to facilitate the understanding of genome functions. a cre/loxp-mediated deletion system for large-scale genome rearrangements in corynebacterium glutamicum was developed. by comparative analysis of c. glutamicum r and c. glutamicum 13032 genomes, distinct 14.5-kb and 56-kb regions not essential for cell survival were identified and targeted for deletion. by homologous recombination, loxp sites were integrated at each end of the target ...	2004	15834716
[metabolic flux analysis of l-valine fermentation in corynebacterium glutamicum].	in industrial fermentation of amino acids the cells are often forced to synthesize the biochemicals excessive of their physiological needs. the knowledge of metabolic networks and their regulation relevant usually come from biochemical research, especially from enzymology, not from engineering study. to enrich the knowledge of metabolic sub-network of l-valine syntheses for higher production of l-valine, corynebacterium glutamicum as1.495 and its genetic derivatives aa361, aat231, aatv341 were u ...	2004	15971614
corynebacterium glutamicum glyceraldehyde-3-phosphate dehydrogenase isoforms with opposite, atp-dependent regulation.	corynebacterium glutamicum gapa and gapb encode glyceraldehyde-3-phosphate dehydrogenases (gapdhs) that differ in molecular weight and activity in the presence of atp. comparative genome analysis revealed that gapa, the product of gapa, represented the canonical gapdh that is highly conserved across the three major life forms. gapb, with an additional 110-residue-long sequence upstream of its gapdh-specific domain, was homologous only to select microbial putative gapdhs. upon gene disruption, th ...	2004	15925900
metabolic engineering of corynebacterium glutamicum for fuel ethanol production under oxygen-deprivation conditions.	the central metabolic pathway of corynebacterium glutamicum was engineered to produce ethanol. a recombinant strain which expressed the zymomonas mobilis genes coding for pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhb) was constructed. both genes placed under the control of the c. glutamicum ldha promoter were expressed at high levels in c. glutamicum, resulting, under oxygen-deprivation conditions, in a significant yield ofethanol from glucose in a process characterized by the abs ...	2004	16179801
the pep-pyruvate-oxaloacetate node as the switch point for carbon flux distribution in bacteria.	in many organisms, metabolite interconversion at the phosphoenolpyruvate (pep)-pyruvate-oxaloacetate node involves a structurally entangled set of reactions that interconnects the major pathways of carbon metabolism and thus, is responsible for the distribution of the carbon flux among catabolism, anabolism and energy supply of the cell. while sugar catabolism proceeds mainly via oxidative or non-oxidative decarboxylation of pyruvate to acetyl-coa, anaplerosis and the initial steps of gluconeoge ...	2004	16102602
seed-specific expression of a bacterial phosphoenolpyruvate carboxylase in vicia narbonensis increases protein content and improves carbon economy.	an ambitious aim in plant breeding and biotechnology is to increase the protein content of crop seeds used for food and feed. using an approach to manipulate assimilate partitioning, we succeeded in elevating the protein content in legume seeds up to 50%. transgenic bean plants were generated which express a corynebacterium glutamicum phosphoenolpyruvate carboxylase (pepc) in a seed-specific manner. the bacterial enzyme was not feedback inhibited by malate. transgenic seeds showed a higher [14c] ...	2004	17147612
lead biosorption by waste biomass of corynebacterium glutamicum generated from lysine fermentation process.	biomass waste, mainly corynebacterium glutamicum, is generated from large-scale lysine fermentation process. in this study, protonated c. glutamicum biomass was evaluated as a biosorbent for the removal of lead from synthetic wastewater. as pb2+ were bound to the biomass, the solution ph deceased, indicating that protons in the biomass were exchanged with lead ions. the corynebacterium biomass bound pb2+ at up to 2.74 mmol g(-1) at ph 5, where lead does not precipitate. compared with other bioso ...	2004	15055771
cloning of the o-acetylhomoserine sulfhydrylase gene from the ruminal bacterium selenomonas ruminantium hd4.	the o-acetylhomoserine sulfhydrylase (oahs) gene was cloned from a selenomonas ruminantium hd4 lambda zap ii genomic library by degenerative probe hybridization and complementation. sequence analysis revealed an 869-bp orf with a g + c content of 53%. the orf had significant homology with enzymes involved in homocysteine biosynthesis. a curablastn homology search showed that the orf has 63% nucleotide identity with the oahs of bacillus stearothermophilus, corynebacterium glutamicum, and acremoni ...	2004	15057458
characterization and chromosomal organization of the murd-murc-ftsq region of corynebacterium glutamicum atcc 13869.	the sequence of a 4.6-kb region of dna from corynebacterium glutamicum atcc 13869 lying upstream from the ftsq-ftsz region has been determined. the region contains four genes with high similarity to the murd, ftsw, murg, and murc genes from different microorganisms. the products of these mur genes probably catalyse several steps in the formation of the precursors for peptidoglycan synthesis in c. glutamicum, whereas ftsw might play also a role in the stabilisation of the ftsz ring during cell di ...	2004	15059630
cation specificity of osmosensing by the betaine carrier betp of corynebacterium glutamicum.	the na(+)/betaine carrier betp from corynebacterium glutamicum was purified and reconstituted in escherichia coli phospholipid liposomes and its osmosensory properties were studied with respect to the cation specificity of osmotic activation. to dissect the influence of the co-substrate na(+) on the energetics of uptake from its possible role as a putative trigger of osmolality-dependent betp activation, the internal na(+) concentration was varied without changing deltapna(+). studying betaine u ...	2004	15063732
production of a novel polygalacturonic acid bioflocculant rea-11 by corynebacterium glutamicum.	the production of a novel polygalacturonic acid bioflocculant rea-11 from a newly isolated strain, corynebacterium glutamicum cctcc m201005, was investigated. sucrose was chosen as a carbon source for rea-11 production. complex nitrogen sources containing urea and an organic nitrogen compound enhanced both bacterial growth and rea-11 production, among which urea plus corn steep liquor was shown to be the most efficient combination. a cost-effective medium for rea-11 production mainly comprised 1 ...	2004	15081493
ramb, a novel transcriptional regulator of genes involved in acetate metabolism of corynebacterium glutamicum.	the adaptation of corynebacterium glutamicum to acetate as a carbon and energy source involves transcriptional regulation of the pta-ack operon coding for the acetate-activating enzymes phosphotransacetylase and acetate kinase and of the acea and aceb genes coding for the glyoxylate cycle enzymes isocitrate lyase and malate synthase, respectively. deletion and mutation analysis of the respective promoter regions led to the identification of highly conserved 13-bp motifs (aa/gaactttgcaaa) as cis- ...	2004	15090522
heterologous expression of lactose- and galactose-utilizing pathways from lactic acid bacteria in corynebacterium glutamicum for production of lysine in whey.	the genetic determinants for lactose utilization from lactobacillus delbrueckii subsp. bulgaricus atcc 11842 and galactose utilization from lactococcus lactis subsp. cremoris mg 1363 were heterologously expressed in the lysine-overproducing strain corynebacterium glutamicum atcc 21253. the c. glutamicum strains expressing the lactose permease and beta-galactosidase genes of l. delbrueckii subsp. bulgaricus exhibited beta-galactosidase activity in excess of 1000 miller units/ml of cells and were ...	2004	15128544
glutamate as an inhibitor of phosphoenolpyruvate carboxylase activity in corynebacterium glutamicum.	the glutamate-producing bacterium, corynebacterium glutamicum is known to possess two anaplerotic enzymes: pyruvate carboxylase (pc) and phosphoenolpyruvate carboxylase (pepc). in vitro, this latter enzyme appeared to be inhibited by different glutamic acid salts, whereas ammonium-glutamate had no influence on pc activity. to investigate the in vivo relevance of pepc activity inhibition, the intracellular concentration of glutamate was determined throughout the glutamate-producing process. the i ...	2004	15133716
the c-terminal domain of the betaine carrier betp of corynebacterium glutamicum is directly involved in sensing k+ as an osmotic stimulus.	the glycine betaine carrier betp of corynebacterium glutamicum was recently shown to function both as an osmosensor and as an osmoregulator in proteoliposomes by sensing changes in the internal k(+) concentration as a measure of hyperosmotic stress. in vivo analysis of mutants carrying deletions at the c-terminal extension of betp indicated that this domain participates in osmostress-dependent activity regulation. to address the question, whether a putative k(+) sensor is located within the c-te ...	2004	15134432
utilization of creatinine as an alternative nitrogen source in corynebacterium glutamicum.	in order to utilize different nitrogen sources and to survive situations of nitrogen limitation, microorganisms have developed several mechanisms to adapt their metabolism to changes in the nitrogen supply. in this communication, the use of creatinine as an alternative nitrogen source in corynebacterium glutamicum, the identification of a membrane protein involved in creatinine uptake, the transcriptional regulation of the corresponding gene, and expression regulation of the gene encoding the cr ...	2004	15148566
identification and characterization of glxr, a gene involved in regulation of glyoxylate bypass in corynebacterium glutamicum.	a corynebacterial clone, previously isolated by scoring repression of laczya fused to the aceb promoter of corynebacterium glutamicum, was analyzed further. in the clone, an open reading frame designated glxr, consisting of 681 nucleotides and encoding a 24,957-da protein, was found. the molecular mass of a native glxr protein was estimated by gel filtration column chromatography to be 44,000 da, suggesting that the protein formed dimers. the predicted amino acid sequence contained both cyclic a ...	2004	15150232
evidence for an arginine exporter encoded by ygga (argo) that is regulated by the lysr-type transcriptional regulator argp in escherichia coli.	the anonymous open reading frame ygga of escherichia coli was identified in this study as a gene that is under the transcriptional control of argp (previously called icia), which encodes a lysr-type transcriptional regulator protein. strains with null mutations in either ygga or argp were supersensitive to the arginine analog canavanine, and ygga-lac expression in vivo exhibited argp(+)-dependent induction by arginine. lysine supplementation phenocopied the argp null mutation in that it virtuall ...	2004	15150242
high level expression of streptomyces mobaraensis transglutaminase in corynebacterium glutamicum using a chimeric pro-region from streptomyces cinnamoneus transglutaminase.	we previously observed secretion of native-type streptomyces mobaraensis transglutaminase (mtgase) in corynebacterium glutamicum by co-expressing the subtilisin-like protease sam-p45 from s. albogriseolus which processes the pro-region. in the present study, we have used a chimeric pro-region consisting of s. mobaraensis and streptomyces cinnamoneus transglutaminases for the production of mtgase in c. glutamicum. as a result, secretion of mtgase using the chimeric pro-region is increased compare ...	2004	15163512
evolutionary process of amino acid biosynthesis in corynebacterium at the whole genome level.	corynebacterium glutamicum, which is the closest relative of corynebacterium efficiens, is widely used for the large scale production of many kinds of amino acids, particularly glutamic acid and lysine, by fermentation. corynebacterium diphtheriae, which is well known as a human pathogen, is also closely related to these two species of corynebacteria, but it lacks such productivity of amino acids. it is an important and interesting question to ask how those closely related bacterial species have ...	2004	15163767
streptomyces lividans and brevibacterium lactofermentum as heterologous hosts for the production of x22 xylanase from aspergillus nidulans.	the aspergillus nidulans gene xlna coding for the fungal xylanase x22 has been cloned and expressed in two heterologous bacterial hosts: streptomyces lividans and brevibacterium lactofermentum. streptomyces strains yielded 10 units/ml of xylanase when the protein was produced with its own signal peptide, and 19 units/ml when its signal peptide was replaced by the one for xylanase xys1 from streptomyces halstedii. b. lactofermentum was also able to produce xylanase x22, affording 6 units/ml upon ...	2004	15168093
functional identification of the gene locus (ncg12319 and characterization of catechol 1,2-dioxygenase in corynebacterium glutamicum.	corynebacterium glutamicum assimilated phenol, benzoate, 4-hydroxybenzoate p-cresol and 3,4-dihydroxybenzoate. ring cleavage was by catechol 1,2-dioxygenase when phenol or benzoate was used and by protocatechuate 3,4-dioxygenase when the others were used as substrate. the locus ncg12319 of its genome was cloned and expressed in escherichia coli. enzyme assays showed that ncg12319 encodes a catechol 1,2-dioxygenase. this catechol 1,2-dioxygenase was purified and accepted catechol, 3-, or 4-methyl ...	2004	15168857
metabolic network analysis of lysine producing corynebacterium glutamicum at a miniaturized scale.	we present a straightforward approach comprising (13)c tracer experiments at 200-microl volume in 96-well microtiter plates with on-line measurement of dissolved oxygen for quantitative high-throughput metabolic network analysis at a miniaturized scale. this method was successfully applied for cultivation and (13)c metabolic flux analysis of two mutants of lysine producing corynebacterium glutamicum (atcc 13287 and atcc 21543). microtiter-plate cultivations showed excellent accordance in kinetic ...	2004	15211482
biochemical and molecular characterization of a ring fission dioxygenase with the ability to oxidize (substituted) salicylate(s) from pseudaminobacter salicylatoxidans.	the gene coding for a dioxygenase with the ability to cleave salicylate by a direct ring fission mechanism to 2-oxohepta-3,5-dienedioic acid was cloned from pseudaminobacter salicylatoxidans strain bn12. the deduced amino acid sequence encoded a protein with a molecular mass of 41,176 da, which showed 28 and 31% sequence identity, respectively, to a gentisate 1,2-dioxygenase from pseudomonas alcaligenes ncimb 9867 and a 1-hydroxy-2-naphthoate 1,2-dioxygenase from nocardioides sp. kp7. the highes ...	2004	15220336
inhibitor-associated transposition events in corynebacterium glutamicum.	in up to 100% of all bacteria grown in the presence of initially inhibitory concentrations of five diverse inhibitors, an extra copy of the resident insertion element is 31831 was found in specific chromosomal regions, the sites of which apparently depended on the inhibitor used. thus, in nine out of nine independently isolated cyanide-associated transpositions, the acquired copy was located within an orf encoding a protein related to the hypothetical but conserved protein yeih of escherichia co ...	2004	15221457
three-dimensional models and structure analysis of corynemycolyltransferases in corynebacterium glutamicum and corynebacterium efficiens.	the corynemycolyltransferase proteins were identified from corynebacterium glutamicum and corynebacterium efficiens genomes using computational tools available in the public domain. three-dimensional models were constructed for corynemycolyltransferases based on the crystal structures of related mycolyltransferases in mycobacterium tuberculosis using the comparative modeling methods. the corynemycolyltransferases share overall an alpha/beta-fold characteristic of the mycolyltransferases despite ...	2004	15225990
transcriptional analysis of the groes-groel1, groel2, and dnak genes in corynebacterium glutamicum: characterization of heat shock-induced promoters.	the appropriate conditions to switch on the heat shock promoters in corynebacterium glutamicum were defined by northern blot analysis. transcriptional patterns were characterized for the groel2 gene and the groes-groel1 and dnak operons. transcriptional start points of these genes were determined by primer extension analysis, allowing the identification of circe and hair boxes close to the -10 and -35 regions of the promoters. the presence of both circe and hair sequences within a single promote ...	2004	15231814
impact of heterologous expression of escherichia coli udp-glucose pyrophosphorylase on trehalose and glycogen synthesis in corynebacterium glutamicum.	trehalose is a disaccharide with a wide range of applications in the food industry. we recently proposed a strategy for trehalose production based on improved strains of the gram-positive bacterium corynebacterium glutamicum. this microorganism synthesizes trehalose through two major pathways, otsba and treyz, by using udp-glucose and adp-glucose, respectively, as the glucosyl donors. in this paper we describe improvement of the udp-glucose supply through heterologous expression in c. glutamicum ...	2004	15240254
osmotic stress response: quantification of cell maintenance and metabolic fluxes in a lysine-overproducing strain of corynebacterium glutamicum.	osmotic stress diminishes cell productivity and may cause cell inactivation in industrial fermentations. the quantification of metabolic changes under such conditions is fundamental for understanding and describing microbial behavior during bioprocesses. we quantified the gradual changes that take place when a lysine-overproducing strain of corynebacterium glutamicum is grown in continuous culture with saline gradients at different dilution rates. the use of compatible solutes depended on enviro ...	2004	15240305
identification of the anabaena sp. strain pcc7120 cyanophycin synthetase as suitable enzyme for production of cyanophycin in gram-negative bacteria like pseudomonas putida and ralstonia eutropha.	the cyanophycin synthetase gene cpha1 encoding the major cyanophycin synthetase (cpha) of anabaena sp. strain pcc7120 was expressed in escherichia coli conferring so far the highest specific cpha activity to e. coli (6.7 nmol arginine per min and mg protein). cpha1 and cpha genes of synechocystis sp. strains pcc6803 and pcc6308 and synechococcus strain ma19 were also expressed in wild types and polyhydroxyalkanoate-negative (pha) mutants of pseudomonas putida and ralstonia eutropha. recombinant ...	2004	15244482
purification and characterization of o-acetylserine sulfhydrylase of corynebacterium glutamicum.	we highly purified o-acetylserine sulfhydrylase from the glutamate-producing bacterium corynebacterium glutamicum. the molecular mass of the purified enzyme was 34,500 as determined by sds-polyacrylamide gel electrophoresis, and 70,800 as determined by gel filtration chromatography. it had an apparent km of 7.0 mm for o-acetylserine and a vmax of 435 micromol min-1 (mg x protein)-1. this is the first report of the cysteine biosynthetic enzyme of c. glutamicum in purified form.	2004	15277766
betp of corynebacterium glutamicum, a transporter with three different functions: betaine transport, osmosensing, and osmoregulation.	in order to circumvent deleterious effects of hypo- and hyperosmotic conditions in its environment, corynebacterium glutamicum has developed a number of mechanisms to counteract osmotic stress. the first response to an osmotic upshift is the activation of uptake mechanisms for the compatible solutes betaine, proline, or ectoine, namely betp, ectp, prop, lcop and putp. betp, the most important uptake system responds to osmotic stress by regulation at the level of both protein activity and gene ex ...	2004	15282171
classification of hyper-variable corynebacterium glutamicum surface-layer proteins by sequence analyses and atomic force microscopy.	the structural s-layer proteins of 28 different corynebacterium glutamicum isolates have been analyzed systematically. treatment of whole c. glutamicum cells with detergents resulted in the isolation of s-layer proteins with different apparent molecular masses, ranging in size from 55 to 66 kda. the s-layer genes analyzed were characterized by coding regions ranging from 1,473 to 1,533 nucleotides coding for s-layer proteins with a size of 490-510 amino acids. using pcr techniques, the correspon ...	2004	15288952
genomewide expression analysis in amino acid-producing bacteria using dna microarrays.	dna microarray technology has become an important research tool for biotechnology and microbiology. it is now possible to characterize genetic diversity and gene expression in a genomewide manner. dna microarrays have been applied extensively to study the biology of many bacteria including escherichia coli, but only recently have they been developed for the gram-positive corynebacterium glutamicum. both bacteria are widely used for biotechnological amino acid production. in this article, in addi ...	2004	15304751
acyl-coa carboxylases (accd2 and accd3), together with a unique polyketide synthase (cg-pks), are key to mycolic acid biosynthesis in corynebacterianeae such as corynebacterium glutamicum and mycobacterium tuberculosis.	the corynebacterianeae such as corynebacterium glutamicum and mycobacterium tuberculosis possess several unique and structurally diverse lipids, including the genus-specific mycolic acids. although the function of a number of genes involved in fatty acid and mycolic acid biosynthesis is known, information relevant to the initial steps within these biosynthetic pathways is relatively sparse. interestingly, the genomes of corynebacterianeae possess a high number of accd genes, whose gene products ...	2004	15308633
lcop, an osmoregulated betaine/ectoine uptake system from corynebacterium glutamicum.	in corynebacterium glutamicum, four uptake systems for compatible solutes have been characterized so far. dhpe (deltabetpdeltaputpdeltapropdeltaectp), a derivative of the c. glutamicum type strain atcc 13032 carrying deletions in the corresponding genes, still showed a low betaine uptake rate of 1.4 nmol/(min mg cdm). genome analyses revealed the presence of a putative carrier, named low capacity osmoregulated permease (lcop), which shows similarities to compatible solute transporters of the bet ...	2004	15327991
comparative genomics identified two conserved dna modules in a corynebacterial plasmid family present in clinical isolates of the opportunistic human pathogen corynebacterium jeikeium.	investigation of 62 clinical isolates of the opportunistic human pathogen corynebacterium jeikeium revealed that 17 possessed plasmids ranging in size from 7.6 to 14.9 kb. the plasmids formed four groups on dna restriction analysis. the complete nucleotide sequence of a representative from each group (pk43, pk64, pcj84, and pb85766) was subsequently determined. additionally, two plasmids (pco455 and pco420) were shown to be derivatives of pk43 and pk64 carrying insertion sequences of the is3 fam ...	2004	15336488
roles of pyruvate kinase and malic enzyme in corynebacterium glutamicum for growth on carbon sources requiring gluconeogenesis.	in many bacteria, pyruvate kinase serves a well-defined function in glycolysis, catalyzing an atp-generating reaction. however, its role during growth on carbon sources requiring glucoeneogenesis is less well investigated. we analyzed a defined pyruvate kinase gene (pyk) deletion mutant of corynebacterium glutamicum, which is unable to grow on ribose as sole carbon source. unexpectedly, the pyk deletion mutant was also unable to grow on acetate or citrate as sole carbon sources unless low amount ...	2004	15375646
comparative metabolic flux analysis of lysine-producing corynebacterium glutamicum cultured on glucose or fructose.	a comprehensive approach to (13)c tracer studies, labeling measurements by gas chromatography-mass spectrometry, metabolite balancing, and isotopomer modeling, was applied for comparative metabolic network analysis of lysine-producing corynebacterium glutamicum on glucose or fructose. significantly reduced yields of lysine and biomass and enhanced formation of dihydroxyacetone, glycerol, and lactate in comparison to those for glucose resulted on fructose. metabolic flux analysis revealed drastic ...	2004	14711646
a polyketide synthase catalyzes the last condensation step of mycolic acid biosynthesis in mycobacteria and related organisms.	mycolic acids are major and specific constituents of the cell envelope of corynebacterineae, a suborder of bacterial species including several important human pathogens such as mycobacterium tuberculosis, mycobacterium leprae, or corynebacterium diphtheriae. these long-chain fatty acids are involved in the unusual architecture and impermeability of the cell envelope of these bacteria. the condensase, the enzyme responsible for the final condensation step in mycolic acid biosynthesis, has remaine ...	2004	14695899
fluorescent phospholipid analogs as microscopic probes for detection of the mycolic acid-containing layer in corynebacterium glutamicum: detecting alterations in the mycolic acid-containing layer following ethambutol treatment.	corynebacterium glutamicum belongs to the mycolic acid-containing actinomycetes, which also include mycobacterium, nocardia, and rhodococcus. the cells of this group possess a cell wall with a thick outer layer composed primarily of mycolic acid, which functions as a permeability barrier. to investigate the mechanism of mycolic acid-containing layer (mycolate layer) formation, we have developed a fluorescence microscopic technique detecting the mycolate layer in situ. the staining specificity of ...	2005	16306684
the clgr protein regulates transcription of the clpp operon in bifidobacterium breve ucc 2003.	five clp genes (clpc, clpb, clpp1, clpp2, and clpx), representing chaperone- and protease-encoding genes, were previously identified in bifidobacterium breve ucc 2003. in the present study, we characterize the b. breve ucc 2003 clpp locus, which consists of two paralogous genes, designated clpp1 and clpp2, whose deduced protein products display significant similarity to characterized clpp peptidases. transcriptional analyses showed that the clpp1 and clpp2 genes are transcribed in response to mo ...	2005	16321946
class 1 integrons and tetracycline resistance genes in alcaligenes, arthrobacter, and pseudomonas spp. isolated from pigsties and manured soil.	the presence of tetracycline resistance (tc(r)) genes and class i integrons (in-1), and their ability to cotransfer were investigated in tc(r) gram-negative (185 strains) and gram-positive (72 strains) bacteria from danish farmland and pigsties. the isolates belonged to the groups or species escherichia coli, enterobacter spp., arthrobacter spp., alcaligenes spp., pseudomonas spp., and corynebacterium glutamicum. the 257 isolates were screened for in-1. eighty-one of the gram-negative isolates w ...	2005	16332771
new multiple-deletion method for the corynebacterium glutamicum genome, using a mutant lox sequence.	due to the difficulty of multiple deletions using the cre/loxp system, a simple, markerless multiple-deletion method based on a cre/mutant lox system combining a right-element (re) mutant lox site with a left-element (le) mutant lox site was employed for large-scale genome rearrangements in corynebacterium glutamicum. eight distinct genomic regions that had been identified previously by comparative analysis of c. glutamicum r and c. glutamicum 13032 genomes were targeted for deletion. by homolog ...	2005	16332837
amplified expression of fructose 1,6-bisphosphatase in corynebacterium glutamicum increases in vivo flux through the pentose phosphate pathway and lysine production on different carbon sources.	the overexpression of fructose 1,6-bisphosphatase (fbpase) in corynebacterium glutamicum leads to significant improvement of lysine production on different sugars. amplified expression of fbpase via the promoter of the gene encoding elongation factor tu (eftu) increased the lysine yield in the feedback-deregulated lysine-producing strain c. glutamicum lyscfbr by 40% on glucose and 30% on fructose or sucrose. additionally formation of the by-products glycerol and dihydroxyacetone was significantl ...	2005	16332851
e1 enzyme of the pyruvate dehydrogenase complex in corynebacterium glutamicum: molecular analysis of the gene and phylogenetic aspects.	the e1p enzyme is an essential part of the pyruvate dehydrogenase complex (pdhc) and catalyzes the oxidative decarboxylation of pyruvate with concomitant acetylation of the e2p enzyme within the complex. we analyzed the corynebacterium glutamicum acee gene, encoding the e1p enzyme, and constructed and characterized an e1p-deficient mutant. sequence analysis of the c. glutamicum acee gene and adjacent regions revealed that acee is not flanked by genes encoding other enzymes of the pdhc. transcrip ...	2005	16109942
identification and characterization of porh, a new cell wall channel of corynebacterium glutamicum.	the cell wall of corynebacterium glutamicum contains the cation-selective channel (porin) pora(c.glut) and the anion-selective channel porb(c.glut) for the passage of hydrophilic solutes. lipid bilayer experiments with organic solvent extracts of whole c. glutamicum cells cultivated in minimal medium suggested that also another cation-selective channel-forming protein, named porh(c.glut), is present in c. glutamicum. the protein was purified to homogeneity by fast-protein liquid chromatography a ...	2005	16112217
enhanced glutamic acid production by a h+-atpase-defective mutant of corynebacterium glutamicum.	previously we reported that a mutant of corynebacterium glutamicum atcc14067 with reduced h+-atpase activity, f172-8, showed an approximately two times higher specific rate of glucose consumption than the parent, but no glutamic acid productivity under the standard biotin-limited culture conditions, where biotin concentration was set at 5.5 microg/l in the production medium (sekine et al., appl. microbiol. biotechnol., 57, 534-540 (2001)). in this study, various culture conditions were tested to ...	2005	16116273
functional genomics and expression analysis of the corynebacterium glutamicum fpr2-cysixhdnyz gene cluster involved in assimilatory sulphate reduction.	corynebacterium glutamicum is a high-gc gram-positive soil bacterium of great biotechnological importance for the production of amino acids. to facilitate the rational design of sulphur amino acid-producing strains, the pathway for assimilatory sulphate reduction providing the necessary reduced sulfur moieties has to be known. although this pathway has been well studied in gram-negative bacteria like escherichia coli and low-gc gram-positives like bacillus subtilis, little is known for the actin ...	2005	16159395
the cgl2612 protein from corynebacterium glutamicum is a drug resistance-related transcriptional repressor: structural and functional analysis of a newly identified transcription factor from genomic dna analysis.	the emergence of antibiotic-resistant bacteria often causes serious clinical problems. the tetr family is one of the major transcription factor families that regulate expression of genes involved in bacterial antimicrobial resistance systems. cgl2612 protein is a transcription factor newly identified by genomic dna analysis on corynebacterium glutamicum, which belongs to the mycolic acid-containing actinomycetales, including the well known pathogens corynebacterium diphtheriae and mycobacterium ...	2005	16166084
the arac-type regulator ripa represses aconitase and other iron proteins from corynebacterium under iron limitation and is itself repressed by dtxr.	the mrna level of the aconitase gene acn of corynebacterium glutamicum is reduced under iron limitation. here we show that an arac-type regulator, termed ripa for "regulator of iron proteins a," is involved in this type of regulation. a c. glutamicum deltaripa mutant has a 2-fold higher aconitase activity than the wild type under iron limitation, but not under iron excess. comparison of the mrna profiles of the deltaripa mutant and the wild type revealed that the acn mrna level was increased in ...	2005	16179344
functional analysis of all aminotransferase proteins inferred from the genome sequence of corynebacterium glutamicum.	twenty putative aminotransferase (at) proteins of corynebacterium glutamicum, or rather pyridoxal-5'-phosphate (plp)-dependent enzymes, were isolated and assayed among others with l-glutamate, l-aspartate, and l-alanine as amino donors and a number of 2-oxo-acids as amino acceptors. one outstanding at identified is alat, which has a broad amino donor specificity utilizing (in the order of preference) l-glutamate > 2-aminobutyrate > l-aspartate with pyruvate as acceptor. another at is avta, which ...	2005	16267288
metabolic engineering of corynebacterium glutamicum for l-serine production.	although l-serine proceeds in just three steps from the glycolytic intermediate 3-phosphoglycerate, and as much as 8% of the carbon assimilated from glucose is directed via l-serine formation, previous attempts to obtain a strain producing l-serine from glucose have not been successful. we functionally identified the genes serc and serb from corynebacterium glutamicum, coding for phosphoserine aminotransferase and phosphoserine phosphatase, respectively. the overexpression of these genes, togeth ...	2005	16269752
characterization of bacterial drug antiporters homologous to mammalian neurotransmitter transporters.	multidrug transporters are ubiquitous proteins, and, based on amino acid sequence similarities, they have been classified into several families. here we characterize a cluster of archaeal and bacterial proteins from the major facilitator superfamily (mfs). one member of this family, the vesicular monoamine transporter (vmat) was previously shown to remove both neurotransmitters and toxic compounds from the cytoplasm, thereby conferring resistance to their effects. a blast search of the available ...	2005	16237035
the osmotic activation of transporter prop is tuned by both its c-terminal coiled-coil and osmotically induced changes in phospholipid composition.	transporter prop of escherichia coli (propec) senses extracellular osmolality and mediates osmoprotectant uptake when it is rising or high. a replica of the propec c terminus (asp468-arg497) forms an intermolecular alpha-helical coiled-coil. this structure is implicated in the osmoregulation of intact propec, in vivo. like that from corynebacterium glutamicum (propcg), the prop orthologue from agrobacterium tumefaciens (propat) sensed and responded to extracellular osmolality after expression in ...	2005	16239220
structure and function of the betaine uptake system betp of corynebacterium glutamicum: strategies to sense osmotic and chill stress.	the soil bacterium corynebacterium glutamicum has to cope with frequent fluctuations of the external osmolarity and temperature. the consequences of hyperosmotic and chill stress seem to differ, either causing dehydration of the cytoplasm or leading to impairment of cellular functions due to low temperature. nevertheless, a particular type of regulatory response, namely the accumulation of so-called compatible solutes, is induced under both conditions. compatible solutes are known to stabilize t ...	2005	16645311
structure determination of secondary transport proteins by electron crystallography: two-dimensional crystallization of the betaine uptake system betp.	structure determination at high resolution is still a challenge for membrane proteins in general, but in particular for secondary transporters due to their highly dynamic nature. x-ray structures of ten secondary transporters have recently been determined, but a thorough understanding of transport mechanisms necessitates structures at different functional states. electron cryo-microscopy of two-dimensional (2d) crystals offers an alternative to obtain structural information at intermediate resol ...	2005	16645315
the transcriptional regulator ssur activates expression of the corynebacterium glutamicum sulphonate utilization genes in the absence of sulphate.	in a recent study, the putative regulatory gene cg0012 was shown to belong to the regulon of mcbr, a global transcriptional regulator of sulphur metabolism in corynebacterium glutamicum atcc 13032. a deletion of cg0012, now designated ssur (sulphonate sulphur utilization regulator), led to the mutant strain c. glutamicum dk100, which was shown to be blocked in the utilization of sulphonates as sulphur sources. according to dna microarray hybridizations, transcription of the ssu and seu genes, en ...	2005	16194234
regulation of amtr-controlled gene expression in corynebacterium glutamicum: mechanism and characterization of the amtr regulon.	amtr, the master regulator of nitrogen control in corynebacterium glutamicum, represses transcription of a number of genes during nitrogen surplus. repression is released by an interaction of amtr with signal transduction protein glnk. as shown by pull-down assays and gel retardation experiments, only adenylylated glnk, which is present in the cells during nitrogen limitation, is able to bind to amtr. the amtr regulon was characterized in this study by a combination of bioinformatics, transcript ...	2005	16194241
biotechnological production of amino acids and derivatives: current status and prospects.	for almost 50 years now, biotechnological production processes have been used for industrial production of amino acids. market development has been particularly dynamic for the flavor-enhancer glutamate and the animal feed amino acids l: -lysine, l: -threonine, and l: -tryptophan, which are produced by fermentation processes using high-performance strains of corynebacterium glutamicum and escherichia coli from sugar sources such as molasses, sucrose, or glucose. but the market for amino acids in ...	2005	16195792
characterization of a corynebacterium glutamicum lactate utilization operon induced during temperature-triggered glutamate production.	gene expression changes of glutamate-producing corynebacterium glutamicum were identified in transcriptome comparisons by dna microarray analysis. during glutamate production induced by a temperature shift, c. glutamicum strain 2262 showed significantly higher mrna levels of the ncgl2816 and ncgl2817 genes than its non-glutamate-producing derivative 2262np. reverse transcription-pcr analysis showed that the two genes together constitute an operon. ncgl2816 putatively codes for a lactate permease ...	2005	16204505
role of the ssu and seu genes of corynebacterium glutamicum atcc 13032 in utilization of sulfonates and sulfonate esters as sulfur sources.	corynebacterium glutamicum atcc 13032 was found to be able to utilize a broad range of sulfonates and sulfonate esters as sulfur sources. the two gene clusters potentially involved in sulfonate utilization, ssud1cba and ssui-seuabc-ssud2, were identified in the genome of c. glutamicum atcc 13032 by similarity searches. while the ssu genes encode proteins resembling ssu proteins from escherichia coli or bacillus subtilis, the seu gene products exhibited similarity to the dibenzothiophene-degradin ...	2005	16204527
analysis of genes involved in arsenic resistance in corynebacterium glutamicum atcc 13032.	corynebacterium glutamicum is able to grow in media containing up to 12 mm arsenite and 500 mm arsenate and is one of the most arsenic-resistant microorganisms described to date. two operons (ars1 and ars2) involved in arsenate and arsenite resistance have been identified in the complete genome sequence of corynebacterium glutamicum. the operons ars1 and ars2 are located some distance from each other in the bacterial chromosome, but they are both composed of genes encoding a regulatory protein ( ...	2005	16204540
the whce gene of corynebacterium glutamicum is important for survival following heat and oxidative stress.	in this study, we have analyzed an orf from corynebacterium glutamicum, which codes for a homologue of the streptomyces coelicolor whib-family of proteins known to be involved in sporulation. this orf encoded a putative protein which harbors a helix-turn-helix dna-binding motif and a probable redox-sensing motif, and has been designated whce. we constructed a whce mutant strain and analyzed the strain under a variety of growth conditions. this mutant strain exhibited a prolonged lag phase and ea ...	2005	16212936
rational design of a corynebacterium glutamicum pantothenate production strain and its characterization by metabolic flux analysis and genome-wide transcriptional profiling.	a "second-generation" production strain was derived from a corynebacterium glutamicum pantothenate producer by rational design to assess its potential to synthesize and accumulate the vitamin pantothenate by batch cultivation. the new pantothenate production strain carries a deletion of the ilva gene to abolish isoleucine synthesis, the promoter down-mutation p-ilvem3 to attenuate ilve gene expression and thereby increase ketoisovalerate availability, and two compatible plasmids to overexpress t ...	2005	15933028
large-scale engineering of the corynebacterium glutamicum genome.	the engineering of corynebacterium glutamicum is important for enhanced production of biochemicals. to construct an improved c. glutamicum genome, we developed a precise genome excision method based on the cre/loxp recombination system and successfully deleted 11 distinct genomic regions identified by comparative analysis of c. glutamicum genomes. despite the loss of several predicted open reading frames, the mutant cells exhibited normal growth under standard laboratory conditions. with a total ...	2005	15933044
dna microarray analysis of the nitrogen starvation response of corynebacterium glutamicum.	nitrogen is an essential component of nearly all of the complex macromolecules in a bacterial cell, e.g. proteins, nucleic acids, and cell wall components. accordingly, most prokaryotes have developed elaborate control mechanisms to provide an optimal supply of nitrogen for cellular metabolism and to cope with situations of nitrogen limitation. in this communication, a global analysis of the corynebacterium glutamicum nitrogen starvation response by transcriptional profiling using dna microarray ...	2005	15935503
the individual and common repertoire of dna-binding transcriptional regulators of corynebacterium glutamicum, corynebacterium efficiens, corynebacterium diphtheriae and corynebacterium jeikeium deduced from the complete genome sequences.	the genus corynebacterium includes gram-positive microorganisms of great biotechnologically importance, such as corynebacterium glutamicum and corynebacterium efficiens, as well as serious human pathogens, such as corynebacterium diphtheriae and corynebacterium jeikeium. although genome sequences of the respective species have been determined recently, the knowledge about the repertoire of transcriptional regulators and the architecture of global regulatory networks is scarce. here, we apply a c ...	2005	15938759
heterologous expression of escherichia coli ppsa (phosphoenolpyruvate synthetase) and galu (udp-glucose pyrophosphorylase) genes in corynebacterium glutamicum, and its impact on trehalose synthesis.	trehalose is a disaccharide with a wide range of applications in the food industry. we recently proposed a strategy for trehalose production based on a corynebacterium glutamicum strain expressing the escherichia coli enzyme udp-glucose pyrophosphorylase (galu). biochemical network analysis suggest a further bottleneck for trehalose synthesis resulting from the coupling of phosphotransferase (pts) mediated glucose uptake, and glucose catabolism in c. glutamicum. to overcome this coupling, we pro ...	2005	15949962
strain improvement by metabolic engineering: lysine production as a case study for systems biology.	a central goal of systems biology is the elucidation of cell function and physiology through the integrated use of broad based genomic and physiological data. such systemic approaches have been employed extensively in the past, as they are a central element of metabolic flux analysis, the distribution of kinetic control in pathways, and the key differentiating characteristic of metabolic engineering. in one case study, these tools have been applied to the improvement of lysine-producing strains ...	2005	15961038
complete genome sequence and analysis of the multiresistant nosocomial pathogen corynebacterium jeikeium k411, a lipid-requiring bacterium of the human skin flora.	corynebacterium jeikeium is a "lipophilic" and multidrug-resistant bacterial species of the human skin flora that has been recognized with increasing frequency as a serious nosocomial pathogen. here we report the genome sequence of the clinical isolate c. jeikeium k411, which was initially recovered from the axilla of a bone marrow transplant patient. the genome of c. jeikeium k411 consists of a circular chromosome of 2,462,499 bp and the 14,323-bp bacteriocin-producing plasmid pkw4. the chromos ...	2005	15968079
vanillate metabolism in corynebacterium glutamicum.	corynebacterium glutamicum, a gram-positive soil bacterium belonging to the mycolic acids-containing actinomycetes, is able to use the lignin degradation products ferulate, vanillate, and protocatechuate as sole carbon sources. the gene cluster responsible for vanillate catabolism was identified and characterized. the vanab genes encoding vanillate demethylase are organized in an operon together with the vank gene, coding for a transport system most likely responsible for protocatechuate uptake. ...	2005	15971090
the transcriptional activator clgr controls transcription of genes involved in proteolysis and dna repair in corynebacterium glutamicum.	expression of the structural genes encoding the atp-dependent proteases clpcp and lon in corynebacterium glutamicum and streptomyces lividans is activated by the transcriptional regulator clgr in response to yet unknown environmental stimuli. as it was not known whether clgr controls expression of additional genes we used dna microarrays in order to comprehensively define the clgr regulon in c. glutamicum. the mrna levels of 16 genes decreased >/= 2-fold in a deltaclgrdeltaclpc mutant (clgr abse ...	2005	15978086
high expression with corynebacterium glutamicum for nuclear magnetic resonance sample preparation.		2005	15979559
lysine and glutamate production by corynebacterium glutamicum on glucose, fructose and sucrose: roles of malic enzyme and fructose-1,6-bisphosphatase.	in the biotechnological production of l-lysine and l-glutamate by corynebacterium glutamicum media based on glucose, fructose or sucrose are typically used. glutamate production by c. glutamicum was very similar on glucose, fructose, glucose plus fructose and sucrose. in contrast, lysine production of genetically defined c. glutamicum strains was significantly higher on glucose than on the other carbon sources. to test whether malic enzyme or fructose-1,6-bisphosphatase might limit growth and ly ...	2005	15979917
application of model discriminating experimental design for modeling and development of a fermentative fed-batch l-valine production process.	a model discriminating experimental design approach for fed-batch processes has been developed and applied to the fermentative production of l-valine by a genetically modified corynebacterium glutamicum strain possessing multiple auxotrophies as an example. being faced with the typical situation of uncertain model information based on preliminary experiments, model discriminating design was successfully applied to improve discrimination between five competing models. within the same modeling and ...	2005	15984033
the amrg1 gene is involved in the activation of acetate in corynebacterium glutamicum.	during growth of corynebacterium glutamicum on acetate as its carbon and energy source, the expression of the pta-ack operon is induced, coding for the acetate-activating enzymes, which are phosphotransacetylase (pta) and acetate kinase (ak). by transposon rescue, we identified the two genes amrg1 and amrg2 found in the deregulated transposon mutant c. glutamicum g25. the amrg1 gene (ncbi-accession: af532964) has a size of 732 bp, encoding a polypeptide of 243 amino acids and apparently is parti ...	2005	15986882
chill activation of compatible solute transporters in corynebacterium glutamicum at the level of transport activity.	the gram-positive soil bacterium corynebacterium glutamicum harbors four osmoregulated secondary uptake systems for compatible solutes, betp, ectp, lcop, and prop. when reconstituted in proteoliposomes, betp was shown to sense hyperosmotic conditions via the increase in luminal k(+) and to respond by instant activation. to study further putative ways of stimulus perception and signal transduction, we have investigated the responses of ectp, lcop, and betp, all belonging to the betaine-carnitine- ...	2005	15995189
two functional fas-i type fatty acid synthases in corynebacterium glutamicum.	the lipid-rich corynebacterianeae, to which corynebacterium glutamicum and mycobacterium species belong, produce both fatty acids and mycolic acids. compared with most other bacteria, c. glutamicum possesses two fatty acid synthases, encoded by fasa (8907 kb; fas-ia) and fasb (8988 kb; fas-ib). here, it was shown by mutational analyses that fasa is essential but fasb is not. however, in a fasa background, the fasb mutation results in a slightly reduced growth yield, l-glutamate production is inc ...	2005	16000732
porh, a new channel-forming protein present in the cell wall of corynebacterium efficiens and corynebacterium callunae.	corynebacterium callunae and corynebacterium efficiens are close relatives of the glutamate-producing mycolata species corynebacterium glutamicum. the properties of the pore-forming proteins, extracted by organic solvents, were studied. the cell extracts contained channel-forming proteins that formed ion-permeable channels with a single-channel conductance of about 2 to 3 ns in 1 m kcl in a lipid bilayer assay. the corresponding proteins from both corynebacteria were purified to homogeneity and ...	2005	16000733
functional identification of novel genes involved in the glutathione-independent gentisate pathway in corynebacterium glutamicum.	corynebacterium glutamicum used gentisate and 3-hydroxybenzoate as its sole carbon and energy source for growth. by genome-wide data mining, a gene cluster designated ncg12918-ncg12923 was proposed to encode putative proteins involved in gentisate/3-hydroxybenzoate pathway. genes encoding gentisate 1,2-dioxygenase (ncg12920) and fumarylpyruvate hydrolase (ncg12919) were identified by cloning and expression of each gene in escherichia coli. the gene of ncg12918 encoding a hypothetical protein (nc ...	2005	16000747
[glyoxylate cycle is required for the overproduction of glutamate but is not essential for corynebacterium glutamicum growth on glucose].	the glyoxylate cycle was hypothesed to be indispensable for glutamate overproduction in coryneform bacteria, for it was thought to fulfill anaplerotic functions and to supply energy during the growth phase. during glutamate overproduction phase, however, it has been noted that the high level of the cycle is detrimental to the glutamate production. in order to clarify the relationship between the glutamate production and the glyoxylate cycle, a chromosomal acea-disrupted mutant of wild-type c. gl ...	2005	16013488
deletion of cg-emb in corynebacterianeae leads to a novel truncated cell wall arabinogalactan, whereas inactivation of cg-ubia results in an arabinan-deficient mutant with a cell wall galactan core.	the cell wall of mycobacterium tuberculosis has a complex ultrastructure that consists of mycolic acids connected to peptidoglycan via arabinogalactan (ag) and abbreviated as the magp complex. the magp complex is crucial for the survival and pathogenicity of m. tuberculosis and is the target of several anti-tubercular agents. apart from sharing a similar magp and the availability of the complete genome sequence, corynebacterium glutamicum has proven useful in the study of orthologous m. tubercul ...	2005	16040600
the impact of fluid mechanical stress on corynebacterium glutamicum during continuous cultivation in an agitated bioreactor.	the effect of mechanical stresses generated by an extreme agitation intensity or a high aeration rate on growth parameters and cell physiology were studied during continuous cultivation of the gram-positive bacterium corynebacterium glutamicum. it is concluded that variations in agitation, aeration rate, or do2 concentrations down to about 1% of saturation do not damage the bacterial cells or cause a significant change in physiological response, as measured by flow cytometry, even though the cel ...	2005	16049736
altered morphology produced by ftsz expression in corynebacterium glutamicum atcc 13869.	corynebacterium glutamicum is a gram-positive bacterium that lacks the cell division ftsa protein and actin-like mreb proteins responsible for determining cylindrical cell shape. when the cell division ftsz gene from c. glutamicum (ftsz(cg)) was cloned in different multicopy plasmids, the resulting constructions could not be introduced into c. glutamicum; it was assumed that elevated levels of ftsz(cg) result in lethality. the presence of a truncated ftsz(cg) and a complete ftsz(cg) under the co ...	2005	16079335
phosphate starvation-inducible gene usha encodes a 5' nucleotidase required for growth of corynebacterium glutamicum on media with nucleotides as the phosphorus source.	phosphorus is an essential component of macromolecules, like dna, and central metabolic intermediates, such as sugar phosphates, and bacteria possess enzymes and control mechanisms that provide an optimal supply of phosphorus from the environment. udp-sugar hydrolases and 5' nucleotidases may play roles in signal transduction, as they do in mammals, in nucleotide salvage, as demonstrated for usha of escherichia coli, or in phosphorus metabolism. the corynebacterium glutamicum gene usha was found ...	2005	16085822
key enzymes of the protocatechuate branch of the beta-ketoadipate pathway for aromatic degradation in corynebacterium glutamicum.	although the protocatechuate branch of the beta-ketoadipate pathway in gram+ bacteria has been well studied, this branch is less understood in gram+ bacteria. in this study, corynebacterium glutamicum was cultivated with protocatechuate, p-cresol, vanillate and 4-hydroxybenzoate as sole carbon and energy sources for growth. enzymatic assays indicated that growing cells on these aromatic compounds exhibited protocatechuate 3,4-dioxygenase activities. data-mining of the genome of this bacterium re ...	2005	16092756
multiarray sensors with pattern recognition for the detection, classification, and differentiation of bacteria at subspecies and strain levels.	this work describes the integration of a fully autonomous electrochemical biosensor with pattern recognition techniques for the detection and classification of bacteria at subspecies and strain level. the system provides a continuous, real-time monitoring of bacteria activity upon exposure to antibiotics. the system utilizes 96-well-type electrodes array (dox-dissolved oxygen sensor) with principal component analysis (pca) for rapid and routine classification of different classes of bacteria and ...	2005	16351141
multiple large segment deletion method for corynebacterium glutamicum.	a precise and scarless genome excision method, employing the cre/loxp system in concert with double-strand break (dsb)-stimulated intramolecular recombination was developed. the dsbs were mediated by the restriction endonuclease, i-scei. it permitted multiple deletions of independent 14-, 43-, and 10-kb-long genomic regions on the corynebacterium glutamicum genome. accuracy of deletion was confirmed by the loss of marker genes, pcr, and sequencing of new genome joints. eleven, 58, and 4 genes we ...	2005	15843930
the mcbr repressor modulated by the effector substance s-adenosylhomocysteine controls directly the transcription of a regulon involved in sulphur metabolism of corynebacterium glutamicum atcc 13032.	in a recent proteomics study we have shown that the mcbr gene of corynebacterium glutamicum atcc 13032 most probably encodes a transcriptional repressor of the tetr type, which regulates the expression of at least six genes involved in the synthesis of sulphur-containing amino acids. by means of dna microarray hybridizations we detected 86 genes with enhanced transcription in an mcbr mutant when compared with the wild-type strain. bioinformatic analysis identified the inverted repeat 5'-tagac-n6 ...	2005	15853877
adaptation of corynebacterium glutamicum to ammonium limitation: a global analysis using transcriptome and proteome techniques.	theresponse of corynebacterium glutamicum to ammonium limitation was studied by transcriptional and proteome profiling of cells grown in a chemostat. our results show that ammonium-limited growth of c. glutamicum results in a rearrangement of the cellular transport capacity, changes in metabolic pathways for nitrogen assimilation, amino acid biosynthesis, and carbon metabolism, as well as a decreased cell division. since transcription at different growth rates was studied, it was possible to dis ...	2005	15870326
ethambutol, a cell wall inhibitor of mycobacterium tuberculosis, elicits l-glutamate efflux of corynebacterium glutamicum.	corynebacterium glutamicum is used for the large-scale production of l-glutamate, but the efflux of this amino acid is poorly understood. this study shows that addition of ethambutol (emb) to growing cultures of c. glutamicum causes l-glutamate efflux at rates of up to 15 nmol min(-1) (mg dry wt)(-1), whereas in the absence of emb, no efflux occurs. emb is used for the treatment of mycobacterium tuberculosis, and at a molecular level it targets a series of arabinosyltransferases (embcab). the si ...	2005	15870446
functional analysis of sigh expression in corynebacterium glutamicum.	the sigh gene of corynebacterium glutamicum encodes ecf sigma factor sigmah. the gene apparently plays an important role in other stress responses as well as heat stress response. in this study, we found that deleting the sigh gene made c. glutamicum cells sensitive to the thiol-specific oxidant diamide. in the sigh mutant strain, the activity of thioredoxin reductase markedly decreased, suggesting that the trxb gene encoding thioredoxin reductase is probably under the control of sigmah. the exp ...	2005	15883048
purification and characterization of succinate:menaquinone oxidoreductase from corynebacterium glutamicum.	succinate:menaquinone oxidoreductase from corynebacterium glutamicum, a high-g+c, gram-positive bacterium, was purified to homogeneity. the enzyme contained two heme b molecules and three polypeptides with apparent molecular masses of 67, 29 and 23 kda, which corresponded to sdha (flavoprotein), sdhb (iron-sulfur protein), and sdhc (membrane anchor protein), respectively. in non-denaturating polyacrylamide gel electrophoresis, the enzyme migrated as a single band with an apparent molecular mass ...	2005	15883782
characterization of methionine export in corynebacterium glutamicum.	corynebacterium glutamicum is known for its effective excretion of amino acids under particular metabolic conditions. concomitant activities of uptake and excretion systems would create an energy-wasting futile cycle; amino acid export systems are therefore tightly regulated. we have used a dna microarray approach to identify genes for membrane proteins which are overexpressed under conditions of elevated cytoplasmic concentrations of methionine. one of these genes was brnf, coding for the large ...	2005	15901702
the crucial role of trehalose and structurally related oligosaccharides in the biosynthesis and transfer of mycolic acids in corynebacterineae.	trehalose (alpha-d-glucopyranosyl-alpha'-d-glucopyranoside) is essential for the growth of the human pathogen mycobacterium tuberculosis but not for the viability of the phylogenetically related corynebacteria. to determine the role of trehalose in the physiology of these bacteria, the so-called corynebacterineae, mutant strains of corynebacterium glutamicum unable to synthesize trehalose due to the knock-out of the genes of the three pathways of trehalose biosynthesis, were biochemically analyz ...	2005	15901732