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characterization of the rna polymerase α subunit operon from corynebacterium ammoniagenes.the rpoa gene, which encodes the α subunit of rna polymerase, and the surrounding regions were cloned from corynebacterium ammoniagenes (atcc 6872), a parental strain of an industrial nucleotide producer in korea. this region encodes genes for the following proteins (in order): initiation factor if-1, the ribosomal proteins s13, s11 and s4, the α subunit of rna polymerase and the ribosomal protein l17. transcript mapping via reverse transcription polymerase chain reaction demonstrates that if1, ...201222806862
biochemical and molecular characterization of the gentisate transporter genk in corynebacterium glutamicum.gentisate (2,5-dihydroxybenzoate) is a key ring-cleavage substrate involved in various aromatic compounds degradation. corynebacterium glutamicum atcc13032 is capable of growing on gentisate and genk was proposed to encode a transporter involved in this utilization by its disruption in the restriction-deficient mutant res167. its biochemical characterization by uptake assay using [(14)c]-labeled gentisate has not been previously reported.201222808015
significance of the cgl1427 gene encoding cytidylate kinase in microaerobic growth of corynebacterium glutamicum.the cgl1427 gene was previously found to be relevant to the microaerobic growth of corynebacterium glutamicum (ikeda et al. biosci biotechnol biochem 73:2806-2808, 2009). in the present work, cgl1427 was identified as a cytidylate kinase gene (cmk) by homology analysis of its deduced amino acid sequence with that of other bacterial cytidylate kinases (cmp kinases) and on the basis of findings that deletion of cgl1427 results in loss of cmp kinase activity. deletion of the cmk gene significantly ...201322810301
isolated microbial single cells and resulting micropopulations grow faster in controlled environments.singularized cells of pichia pastoris, hansenula polymorpha, and corynebacterium glutamicum displayed specific growth rates under chemically and physically constant conditions that were consistently higher than those obtained in populations. this highlights the importance of single-cell analyses by uncoupling physiology and the extracellular environment, which is now possible using the envirostat 2.0 concept.201222820335
reductive whole-cell biotransformation with corynebacterium glutamicum: improvement of nadph generation from glucose by a cyclized pentose phosphate pathway using pfka and gapa deletion mutants.in this study, the potential of corynebacterium glutamicum for reductive whole-cell biotransformation is shown. the nadph-dependent reduction of the prochiral methyl acetoacetate (maa) to the chiral (r)-methyl 3-hydroxybutyrate (mhb) by an alcohol dehydrogenase from lactobacillus brevis (lbadh) was used as model reaction and glucose served as substrate for the regeneration of nadph. since nadph is mainly formed in the oxidative branch of the pentose phosphate pathway (ppp), c. glutamicum was eng ...201222851018
glucosamine as carbon source for amino acid-producing corynebacterium glutamicum.corynebacterium glutamicum grows with a variety of carbohydrates and carbohydrate derivatives as sole carbon sources; however, growth with glucosamine has not yet been reported. we isolated a spontaneous mutant (m4) which is able to grow as fast with glucosamine as with glucose as sole carbon source. glucosamine also served as a combined source of carbon, energy and nitrogen for the mutant strain. characterisation of the m4 mutant revealed a significantly increased expression of the nagb gene en ...201322854894
altered large-ring cyclodextrin product profile due to a mutation at tyr-172 in the amylomaltase of corynebacterium glutamicum.corynebacterium glutamicum amylomaltase (cgam) catalyzes the formation of large-ring cyclodextrins (lr-cds) with a degree of polymerization of 19 and higher. the cloned cgam gene was ligated into the pet-17b vector and used to transform escherichia coli bl21(de3). site-directed mutagenesis of tyr-172 in cgam to alanine (y172a) was performed to determine its role in the control of lr-cd production. both the recombinant wild-type (wt) and y172a enzymes were purified to apparent homogeneity and cha ...201222865069
increased lysine production by flux coupling of the tricarboxylic acid cycle and the lysine biosynthetic pathway--metabolic engineering of the availability of succinyl-coa in corynebacterium glutamicum.in this study, we demonstrate increased lysine production by flux coupling using the industrial work horse bacterium corynebacterium glutamicum, which was mediated by the targeted interruption of the tricarboxylic acid (tca) cycle at the level of succinyl-coa synthetase. the succinylase branch of the lysine production pathway functions as the bridging reaction to convert succinyl-coa to succinate in this aerobic bacterium. the mutant c. glutamicum δsuccd showed a 60% increase in the yield of lys ...201322871505
construction of in vitro transcription system for corynebacterium glutamicum and its use in the recognition of promoters of different classes.to facilitate transcription studies in corynebacterium glutamicum, we have developed an in vitro transcription system for this bacterium used as an industrial producer of amino acids and a model organism for actinobacteria. this system consists of c. glutamicum rna polymerase (rnap) core (α2, β, β'), a sigma factor and a promoter-carrying dna template, that is specifically recognized by the rnap holoenzyme formed. the rnap core was purified from the c. glutamicum strain with the modified rpoc ge ...201222885668
beyond growth rate 0.6: corynebacterium glutamicum cultivated in highly diluted environments.fast growth of industrial microorganisms, such as corynebacterium glutamicum, is a direct amplifier for the productivity of any growth coupled or decoupled production process. recently, it has been shown that c. glutamicum when grown in a novel picoliter bioreactor (plbr) exhibits a 50% higher growth rate compared to a 1 l batch cultivation [grünberger et al. (2012) lab chip]. we here compare growth of c. glutamicum with glucose as substrate at different scales covering batch cultivations in the ...201322890752
production of l-lysine on different silage juices using genetically engineered corynebacterium glutamicum.corynebacterium glutamicum, the best established industrial producer organism for lysine was genetically modified to allow the production of lysine on grass and corn silages. the resulting strain c. glutamicum lysc(fbr)dld(psod)pyc(psod)male(psod)fbp(psod)gapx(psod) was based on earlier work (neuner and heinzle, 2011). that mutant carries a point mutation in the aspartokinase (lysc) regulatory subunit gene as well as overexpression of d-lactate dehydrogenase (dld), pyruvate carboxylase (pyc) and ...201322898177
transcriptional cross-regulation between gram-negative and gram-positive bacteria, demonstrated using argp-argo of escherichia coli and lysg-lyse of corynebacterium glutamicum.the protein-gene pairs argp-argo of escherichia coli and lysg-lyse of corynebacterium glutamicum are orthologous, with the first member of each pair being a lysr-type transcriptional regulator and the second its target gene encoding a basic amino acid exporter. whereas lyse is an exporter of arginine (arg) and lysine (lys) whose expression is induced by arg, lys, or histidine (his), argo exports arg alone, and its expression is activated by arg but not lys or his. we have now reconstituted in e. ...201222904281
[construction and structural analysis of integrated cellular network of corynebacterium glutamicum].corynebacterium glutamicum is one of the most important traditional industrial microorganisms and receiving more and more attention towards a novel cellular factory due to the recently rapid development in genomics and genetic operation toolboxes for corynebacterium. however, compared to other model organisms such as escherichia coli, there were few studies on its metabolic regulation, especially a genome-scale integrated cellular network model currently missing for corynebacterium, which hinder ...201222916496
sialic acid utilization by the soil bacterium corynebacterium glutamicum.the ability to use the sialic acid, n-acetylneuraminic acid, neu5ac, as a nutrient has been characterized in a number of bacteria, most of which are human pathogens that encounter this molecule because of its presence on mucosal surfaces. the soil bacterium corynebacterium glutamicum also has a full complement of genes for sialic acid catabolism, and we demonstrate that it can use neu5ac as a sole source of carbon and energy and isolate mutants with a much reduced growth lag on neu5ac. disruptio ...201222924979
destabilized eyfp variants for dynamic gene expression studies in corynebacterium glutamicum.fluorescent reporter proteins are widely used for the non-invasive monitoring of gene expression patterns, but dynamic measurements are hampered by the extremely high stability of gfp and homologue proteins. in this study, we used ssra-mediated peptide tagging for the construction of unstable variants of the gfp derivative eyfp (enhanced yellow fluorescent protein) and applied those for transient gene expression analysis in the industrial platform organism corynebacterium glutamicum.201222938655
chemometric formulation of bacterial consortium-avs for improved decolorization of resonance-stabilized and heteropolyaromatic dyes.a bacterial consortium-avs, consisting of pseudomonas desmolyticum ncim 2112, kocuria rosea mtcc 1532 and micrococcus glutamicus ncim 2168 was formulated chemometrically, using the mixture design matrix based on the design of experiments methodology. the formulated consortium-avs decolorized acid blue 15 and methylene blue with a higher average decolorization rate, which is more rapid than that of the pure cultures. the uv-vis spectrophotometric, fourier transform infra red spectrophotometric an ...201222940340
alternating-access mechanism in conformationally asymmetric trimers of the betaine transporter betp.betaine and na(+) symport has been extensively studied in the osmotically regulated transporter betp from corynebacterium glutamicum, a member of the betaine/choline/carnitine transporter family, which shares the conserved leut-like fold of two inverted structural repeats. betp adjusts its transport activity by sensing the cytoplasmic k(+) concentration as a measure for hyperosmotic stress via the osmosensing carboxy-terminal domain. betp needs to be in a trimeric state for communication between ...201222940865
transcriptional regulation of the operon encoding stress-responsive ecf sigma factor sigh and its anti-sigma factor rsha, and control of its regulatory network in corynebacterium glutamicum.the expression of genes in corynebacterium glutamicum, a gram-positive non-pathogenic bacterium used mainly for the industrial production of amino acids, is regulated by seven different sigma factors of rna polymerase, including the stress-responsive ecf-sigma factor sigh. the sigh gene is located in a gene cluster together with the rsha gene, putatively encoding an anti-sigma factor. the aim of this study was to analyze the transcriptional regulation of the sigh and rsha gene cluster and the ef ...201222943411
single-step production of polyhydroxybutyrate from starch by using α-amylase cell-surface displaying system of corynebacterium glutamicum.direct polyhydroxybutyrate (phb) production from starch was for the first time achieved using engineered corynebacterium glutamicum expressing phb biosynthetic genes and displaying α-amylase on its cell surface. the engineered strain accumulated 6.4 wt% phb from starch which was higher than that obtained from glucose (4.9 wt%).201322959444
quantitation of intracellular purine intermediates in different corynebacteria using electrospray lc-ms/ms.intermediates of the purine biosynthesis pathway play key roles in cellular metabolism including nucleic acid synthesis and signal mediation. in addition, they are also of major interest to the biotechnological industry as several intermediates either possess flavor-enhancing characteristics or are applied in medical therapy. in this study, we have developed an analytical method for quantitation of 12 intermediates from the purine biosynthesis pathway including important nucleotides and their co ...201222960872
carotenoid biosynthesis and overproduction in corynebacterium glutamicum.corynebacterium glutamicum contains the glycosylated c50 carotenoid decaprenoxanthin as yellow pigment. starting from isopentenyl pyrophosphate, which is generated in the non-mevalonate pathway, decaprenoxanthin is synthesized via the intermediates farnesyl pyrophosphate, geranylgeranyl pyrophosphate, lycopene and flavuxanthin.201222963379
extensive exometabolome analysis reveals extended overflow metabolism in various microorganisms.overflow metabolism is well known for yeast, bacteria and mammalian cells. it typically occurs under glucose excess conditions and is characterized by excretions of by-products such as ethanol, acetate or lactate. this phenomenon, also denoted the short-term crabtree effect, has been extensively studied over the past few decades, however, its basic regulatory mechanism and functional role in metabolism is still unknown. here we present a comprehensive quantitative and time-dependent analysis of ...201222963408
a propionate-inducible expression system based on the corynebacterium glutamicum prpd2 promoter and prpr activator and its application for the redirection of amino acid biosynthesis pathways.a novel expression system for corynebacterium glutamicum, based on the transcriptional activator of propionate metabolism genes prpr and its target promoter/operator sequence, was developed and tested. the activator prpr is co-activated by propionate added to the growth medium. in a minimal medium a propionate concentration of only 1 mg l⁻¹ was found to be sufficient for full induction (up to 120-fold) of its native target, the propionate metabolism operon prpdbc2. then, an artificial transcript ...201322982516
implication of gluconate kinase activity in l-ornithine biosynthesis in corynebacterium glutamicum.with the purpose of generating a microbial strain for l-ornithine production in corynebacterium glutamicum, genes involved in the central carbon metabolism were inactivated so as to modulate the intracellular level of nadph, and to evaluate their effects on l-ornithine production in c. glutamicum. upon inactivation of the 6-phosphoglucoisomerase gene (pgi) in a c. glutamicum strain, the concomitant increase in intracellular nadph concentrations from 2.55 to 5.75 mmol g⁻¹ (dry cell weight) was ac ...201222987028
coordinated regulation of gnd, which encodes 6-phosphogluconate dehydrogenase, by the two transcriptional regulators gntr1 and rama in corynebacterium glutamicum.the transcriptional regulation of corynebacterium glutamicum gnd, encoding 6-phosphogluconate dehydrogenase, was investigated. two transcriptional regulators, gntr1 and rama, were isolated by affinity purification using gnd promoter dna. gntr1 was previously identified as a repressor of gluconate utilization genes, including gnd. involvement of rama in gnd expression had not been investigated to date. the level of gnd mrna was barely affected by the single deletion of rama. however, gnd expressi ...201223024346
the ppm operon is essential for acylation and glycosylation of lipoproteins in corynebacterium glutamicum.due to their contribution to bacterial virulence, lipoproteins and members of the lipoprotein biogenesis pathway represent potent drug targets. following translocation across the inner membrane, lipoprotein precursors are acylated by lipoprotein diacylglycerol transferase (lgt), cleaved off their signal peptides by lipoprotein signal peptidase (lsp) and, in gram-negative bacteria, further triacylated by lipoprotein n-acyl transferase (lnt). the existence of an active apolipoprotein n-acyltransfe ...201223029442
model-based analysis of an adaptive evolution experiment with escherichia coli in a pyruvate limited continuous culture with glycerol.: bacterial strains that were genetically blocked in important metabolic pathways and grown under selective conditions underwent a process of adaptive evolution: certain pathways may have been deregulated and therefore allowed for the circumvention of the given block. a block of endogenous pyruvate synthesis from glycerol was realized by a knockout of pyruvate kinase and phosphoenolpyruvate carboxylase in e. coli. the resulting mutant strain was able to grow on a medium containing glycerol and l ...201223033959
the two-component system chrsa is crucial for haem tolerance and interferes with hrrsa in haem-dependent gene regulation in corynebacterium glutamicum.we recently showed that the two-component system (tcs) hrrsa plays a central role in the control of haem homeostasis in the gram-positive soil bacterium corynebacterium glutamicum. here, we characterized the function of another tcs of this organism, chrsa, which exhibits significant sequence similarity to hrrsa, and provide evidence for cross-regulation of the two systems. in this study, chrsa was shown to be crucial for haem resistance of c. glutamicum by activation of the putative haem-detoxif ...201223038807
3' untranslated region-dependent degradation of the acea mrna, encoding the glyoxylate cycle enzyme isocitrate lyase, by rnase e/g in corynebacterium glutamicum.we previously reported that the corynebacterium glutamicum rnase e/g encoded by the rneg gene (ncgl2281) is required for the 5' maturation of 5s rrna. in the search for the intracellular target rnas of rnase e/g other than the 5s rrna precursor, we detected that the amount of isocitrate lyase, an enzyme of the glyoxylate cycle, increased in rneg knockout mutant cells grown on sodium acetate as the sole carbon source. rifampin chase experiments showed that the half-life of the acea mrna was about ...201223042181
corynebacterium glutamicum zur acts as a zinc-sensing transcriptional repressor of both zinc-inducible and zinc-repressible genes involved in zinc homeostasis.zur is a zinc-dependent transcriptional repressor of zinc uptake systems in bacteria. in the present study, we examined the role of corynebacterium glutamicum zur in the zinc-inducible expression of two genes: one encoding a cation diffusion facilitator (zrf) and the other a metal-translocating p-type atpase (zra). both genes were shown to be involved in zinc resistance. disruption of the zur gene encoding zur resulted in constitutive expression of zrf and zra mrnas. an electrophoretic mobility ...201223061624
analysis of corynebacterium glutamicum promoters and their applications.promoters are dna sequences which function as regulatory signals of transcription initiation catalyzed by rna polymerase. since promoters substantially influence levels of gene expression, they have become powerful tools in metabolic engineering. methods for their localization used in corynebacterium glutamicum and techniques for the analysis of their function are described in this review. c. glutamicum promoters can be classified according to the respective σ factors which direct rna polymerase ...201223080252
molecular mechanisms and metabolic engineering of glutamate overproduction in corynebacterium glutamicum.glutamate is a commercially important chemical. it is used as a flavor enhancer and is a major raw material for producing industrially useful chemicals. a coryneform bacterium, corynebacterium glutamicum, was isolated in 1956 by japanese researchers as a glutamate-overproducing bacterium and since then, remarkable progress in glutamate production has been made using this microorganism. currently, the global market for glutamate is over 2.5 million tons per year. glutamate overproduction by c. gl ...201223080255
sugar transport systems in corynebacterium glutamicum: features and applications to strain development.corynebacterium glutamicum uses the phosphoenolpyruvate-dependent sugar phosphotransferase system (pts) to take up and phosphorylate glucose, fructose, and sucrose, the major sugars from agricultural crops that are used as the primary feedstocks for industrial amino acid fermentation. this means that worldwide amino acid production using this organism has depended exclusively on the pts. recently, a better understanding not only of pts-mediated sugar uptake but also of global regulation associat ...201223081775
corynebacterium glutamicum csor acts as a transcriptional repressor of two copper/zinc-inducible p(1b)-type atpase operons.the mechanism of regulation of the expression of copa and copb, encoding putative copper-translocating p(1b)-type atpases in corynebacterium glutamicum, was investigated. the levels of copa and copb mrnas were upregulated in response to excess copper as well as excess zinc. disruption of csor, encoding a transcriptional regulator, resulted in constitutive expression of copa and copb. the csor protein bound to the promoter regions of the copa-csor and the cgr_0124-copb-cgr_0126 operon. in vitro d ...201223090582
the lipoprotein lpqw is essential for the mannosylation of periplasmic glycolipids in corynebacteria.phosphatidylinositol mannosides (pim), lipomannan (lm), and lipoarabinomannan (lam) are essential components of the cell wall and plasma membrane of mycobacteria, including the human pathogen mycobacterium tuberculosis, as well as the related corynebacterineae. we have previously shown that the lipoprotein, lpqw, regulates pim and lm/lam biosynthesis in mycobacteria. here, we provide direct evidence that lpqw regulates the activity of key mannosyltransferases in the periplasmic leaflet of the ce ...201223091062
metabolic engineering of the purine biosynthetic pathway in corynebacterium glutamicum results in increased intracellular pool sizes of imp and hypoxanthine.purine nucleotides exhibit various functions in cellular metabolism. besides serving as building blocks for nucleic acid synthesis, they participate in signaling pathways and energy metabolism. further, imp and gmp represent industrially relevant biotechnological products used as flavor enhancing additives in food industry. therefore, this work aimed towards the accumulation of imp applying targeted genetic engineering of corynebacterium glutamicum.201223092390
high-resolution detection of dna binding sites of the global transcriptional regulator glxr in corynebacterium glutamicum.the transcriptional regulator glxr has been characterized as a global hub within the gene-regulatory network of corynebacterium glutamicum. chromatin immunoprecipitation with a specific anti-glxr antibody and subsequent high-throughput sequencing (chip-seq) was applied to c. glutamicum to get new in vivo insights into the gene composition of the glxr regulon. in a comparative approach, c. glutamicum cells were grown with either glucose or acetate as the sole carbon source prior to immunoprecipit ...201323103979
identification of a had superfamily phosphatase, hdpa, involved in 1,3-dihydroxyacetone production during sugar catabolism in corynebacterium glutamicum.corynebacterium glutamicum produces 1,3-dihydroxyacetone (dha) as metabolite of sugar catabolism but the responsible enzyme is yet to be identified. using a transposon mutant library, the gene hdpa (cgr_2128) was shown to encode a haloacid dehalogenase superfamily member that catalyzes dephosphorylation of dihydroxyacetone phosphate to produce dha. inactivation of hdpa led to a drastic decrease in dha production from each of glucose, fructose, and sucrose, indicating that hdpa is the main enzyme ...201223108048
an automated workflow for enhancing microbial bioprocess optimization on a novel microbioreactor platform.high-throughput methods are widely-used for strain screening effectively resulting in binary information regarding high or low productivity. nevertheless achieving quantitative and scalable parameters for fast bioprocess development is much more challenging, especially for heterologous protein production. here, the nature of the foreign protein makes it impossible to predict the, e.g. best expression construct, secretion signal peptide, inductor concentration, induction time, temperature and sub ...201223113930
next-generation sequencing-based genome-wide mutation analysis of l-lysine-producing corynebacterium glutamicum atcc 21300 strain.in order to identify single nucleotide polymorphism and insertion/deletion mutations, we performed whole-genome re-sequencing of the enhanced l-lysine-producing corynebacterium glutamicum atcc 21300 strain. in total, 142 single nucleotide polymorphisms and 477 insertion/deletion mutations were identified in the atcc 21300 strain when compared to 3,434 predicted genes of the wild-type c. glutamicum atcc 13032 strain. among them, 110 transitions and 29 transversions of single nucleotide polymorphi ...201223124757
design and testing of a synthetic biology framework for genetic engineering of corynebacterium glutamicum.synthetic biology approaches can make a significant contribution to the advance of metabolic engineering by reducing the development time of recombinant organisms. however, most of synthetic biology tools have been developed for escherichia coli. here we provide a platform for rapid engineering of c. glutamicum, a microorganism of great industrial interest. this bacteria, used for decades for the fermentative production of amino acids, has recently been developed as a host for the production of ...201223134565
heterologous carotenoid-biosynthetic enzymes: functional complementation and effects on carotenoid profiles in escherichia coli.a limited number of carotenoid pathway genes from microbial sources have been studied for analyzing the pathway complementation in the heterologous host escherichia coli. in order to systematically investigate the functionality of carotenoid pathway enzymes in e. coli, the pathway genes of carotenogenic microorganisms (brevibacterium linens, corynebacterium glutamicum, rhodobacter sphaeroides, rhodobacter capsulatus, rhodopirellula baltica, and pantoea ananatis) were modified to form synthetic e ...201323144136
differential arabinan capping of lipoarabinomannan modulates innate immune responses and impacts t helper cell differentiation.toll-like receptors (tlrs) recognize pathogens by interacting with pathogen-associated molecular patterns, such as the phosphatidylinositol-based lipoglycans, lipomannan (lm) and lipoarabinomannan (lam). such structures are present in several pathogens, including mycobacterium tuberculosis, being important for the initiation of immune responses. it is well established that the interaction of lm and lam with tlr2 is a process dependent on the structure of the ligands. however, the implications of ...201223144457
secretory production of an fad cofactor-containing cytosolic enzyme (sorbitol-xylitol oxidase from streptomyces coelicolor) using the twin-arginine translocation (tat) pathway of corynebacterium glutamicum.carbohydrate oxidases are biotechnologically interesting enzymes that require a tightly or covalently bound cofactor for activity. using the industrial workhorse corynebacterium glutamicum as the expression host, successful secretion of a normally cytosolic fad cofactor-containing sorbitol-xylitol oxidase from streptomyces coelicolor was achieved by using the twin-arginine translocation (tat) protein export machinery for protein translocation across the cytoplasmic membrane. our results demonstr ...201323163932
accelerated pentose utilization by corynebacterium glutamicum for accelerated production of lysine, glutamate, ornithine and putrescine.because of their abundance in hemicellulosic wastes arabinose and xylose are an interesting source of carbon for biotechnological production processes. previous studies have engineered several corynebacterium glutamicum strains for the utilization of arabinose and xylose, however, with inefficient xylose utilization capabilities. to improve xylose utilization, different xylose isomerase genes were tested in c. glutamicum. the gene originating from xanthomonas campestris was shown to have the hig ...201223164409
influence of sigb inactivation on corynebacterium glutamicum protein secretion.the non-essential corynebacterium glutamicum sigma factor, sigb, modulates global gene expression during the transition from exponential growth to the stationary phase. utilizing a signal peptide derived from c. glutamicum r cgr_0949, a sigb disruption mutant able to secrete 3- to 5-fold more green fluorescence protein (gfp) and α-amylase than the wild type strain was isolated. the signal peptide selectively enabled the mutant to produce greater amounts of both proteins, which were in turn secre ...201323179627
ruthenium recovery from acetic acid waste water through sorption with bacterial biosorbent fibers.a fibrous bacterial biosorbent was developed to bind precious metal-organic complexes in batch and column processes. polyethylenimine (pei)-modified bacterial biosorbent fiber (pbbf) was prepared by spinning corynebacterium glutamicum biomass-chitosan blends, coating them with pei and cross-linking with glutaraldehyde. when an acetic acid waste solution containing 1822.9mg/l ru was used as a model waste solution, ru uptake by the pbbf was 16.5 times higher than that of the commercial ion exchang ...201323196218
bio-based production of organic acids with corynebacterium glutamicum.the shortage of oil resources, the steadily rising oil prices and the impact of its use on the environment evokes an increasing political, industrial and technical interest for development of safe and efficient processes for the production of chemicals from renewable biomass. thus, microbial fermentation of renewable feedstocks found its way in white biotechnology, complementing more and more traditional crude oil-based chemical processes. rational strain design of appropriate microorganisms has ...201323199277
analysis of in vivo function of predicted isoenzymes:a metabolomic approach.isoenzymes occur in all organisms. often, they are regulated with respect to environmental perturbations to assure metabolic flexibility. in bioinformatics-driven functional genome annotations, the presence of isoenzymes is frequently predicted. it is desirable to verify the functions of putative isoenzymes experimentally for a correct estimation of the metabolic capacities in a given organism. using metabolome analysis, we investigated two knockout mutants of putative shikimate dehydrogenases ( ...201223215805
picoliter ndep traps enable time-resolved contactless single bacterial cell analysis in controlled microenvironments.we present a lab-on-a-chip device, the envirostat 2.0, which allows for the first time contactless cultivation of a single bacterial cell by negative dielectrophoresis (ndep) in a precisely controllable microenvironment. stable trapping in perfusing growth medium was achieved by a miniaturization of octupole electrode geometries, matching the dimensions of bacteria. temperature sensitive fluorescent measurements showed that these reductions of microelectrode distances led to reduced joule heatin ...201323223864
engineering of corynebacterium glutamicum for high-yield l-valine production under oxygen deprivation conditions.we previously demonstrated efficient l-valine production by metabolically engineered corynebacterium glutamicum under oxygen deprivation. to achieve the high productivity, a nadh/nadph cofactor imbalance during the synthesis of l-valine was overcome by engineering nad-preferring mutant acetohydroxy acid isomeroreductase (ahair) and using nad-specific leucine dehydrogenase from lysinibacillus sphaericus. lactate as a by-product was largely eliminated by disrupting the lactate dehydrogenase gene l ...201323241971
metabolic engineering of industrial platform microorganisms for biorefinery applications--optimization of substrate spectrum and process robustness by rational and evolutive strategies.bio-based production promises a sustainable route to myriads of chemicals, materials and fuels. with regard to eco-efficiency, its future success strongly depends on a next level of bio-processes using raw materials beyond glucose. such renewables, i.e., polymers, complex substrate mixtures and diluted waste streams, often cannot be metabolized naturally by the producing organisms. this particularly holds for well-known microorganisms from the traditional sugar-based biotechnology, including esc ...201323260271
monitoring of population dynamics of corynebacterium glutamicum by multiparameter flow cytometry.phenotypic variation of microbial populations is a well-known phenomenon and may have significant impact on the success of industrial bioprocesses. flow cytometry (fc) and the large repertoire of fluorescent dyes bring the high-throughput analysis of multiple parameters in single bacterial cells into reach. in this study, we evaluated a set of different fluorescent dyes for suitability in fc single cell analysis of the biotechnological platform organism corynebacterium glutamicum. already simple ...201323279937
non-invasive online detection of microbial lysine formation in stirred tank bioreactors by using calorespirometry.non-invasive methods for online monitoring of biotechnological processes without compromising the integrity of the reactor system are very important to generate continuous data. even though calorimetry has been used in conventional biochemical analysis for decades, it has not yet been specifically applied for online detection of product formation at technical scale. thus, this article demonstrates a calorespirometric method for online detection of microbial lysine formation in stirred tank biore ...201323280310
adaptive evolution of corynebacterium glutamicum resistant to oxidative stress and its global gene expression profiling.corynebacterium glutamicum was adapted in a chemostat for 1,900 h with gradually increasing h2o2 stress to understand the oxidative stress response of an industrial host. after 411 generations of adaptation, c. glutamicum developed the ability to grow under stress of 10 mm h2o2, whereas the wild-type did not. the adapted strain also showed increased stress resistance to diamide and menadione, sds, tween 20, hcl, naoh, and ampicillin. a total of 1,180 genes in the rna-seq transcriptome analysis o ...201323288296
[overexpression of corynebacterium glutamicum nad kinase improves l-isoleucine biosynthesis].nad kinase catalyzes the phosphorylation of coenzyme i [nad(h)] to form coenzyme ii [nadp(h)], and nadph is an important cofactor in l-isoleucine biosynthesis. in order to improve nadph supply, ppnk, the gene encoding nad kinase in corynebacterium glutamicum was cloned and separately expressed in an l-isoleucine synthetic strain, brevibacterium lactofermentum jhi3-156, by an inducible expression vector pdxw-8 and a constitutive expression vector pdxw-9. compared with the control strain jhi3-156/ ...201223289306
recent advancements in various steps of ethanol, butanol, and isobutanol productions from woody materials.in this review, the recent advancements and technical challenges associated with the production of ethanol, butanol, and isobutanol via bioconversion routes from celluloses of woody materials are reviewed. physicochemical processes, e.g. steam explosion, seem to be the most viable process for pretreating woody materials. although enzymatic hydrolysis is selective, it is rather a slow process. acid hydrolysis is a relatively fast process with a high yield, but it produces inhibitory compounds of ...201323297056
transcriptional control of the f0f1-atp synthase operon of corynebacterium glutamicum: sigmah factor binds to its promoter and regulates its expression at different ph values.corynebacterium glutamicum used in the amino acid fermentation industries is an alkaliphilic microorganism. its f(0)f(1)-atpase operon (atpbefhagdc) is expressed optimally at ph 9.0 forming a polycistronic (7.5 kb) and a monocistronic (1.2 kb) transcripts both starting upstream of the atpb gene. expression of this operon is controlled by the sigmah factor. the sigmah gene (sigh) was cloned and shown to be co-transcribed with a small gene, cg0877, encoding a putative anti-sigma factor. a mutant d ...201323298179
corynebacterium glutamicum promoters: a practical approach.transcription initiation is the key step in gene expression in bacteria, and it is therefore studied for both theoretical and practical reasons. promoters, the traffic lights of transcription initiation, are used as construction elements in biotechnological efforts to coordinate 'green waves' in the metabolic pathways leading to the desired metabolites. detailed analyses of corynebacterium glutamicum promoters have already provided large amounts of data on their structures, regulatory mechanisms ...201323305350
characterization of shikimate dehydrogenase homologues of corynebacterium glutamicum.the function of three corynebacterium glutamicum shikimate dehydrogenase homologues, designated as qsud (cgr_0495), cgr_1216, and aroe (cgr_1677), was investigated. a disruptant of aroe required shikimate for growth, whereas a qsud-deficient strain did not grow in medium supplemented with either quinate or shikimate as sole carbon sources. there was no discernible difference in growth rate between wild-type and a cgr_1216-deficient strain. enzymatic assays showed that aroe both reduced 3-dehydro ...201323306642
recognition of microbial and mammalian phospholipid antigens by nkt cells with diverse tcrs.cd1d-restricted natural killer t (nkt) cells include two major subgroups. the most widely studied are vα14jα18(+) invariant nkt (inkt) cells that recognize the prototypical α-galactosylceramide antigen, whereas the other major group uses diverse t-cell receptor (tcr) α-and β-chains, does not recognize α-galactosylceramide, and is referred to as diverse nkt (dnkt) cells. dnkt cells play important roles during infection and autoimmunity, but the antigens they recognize remain poorly understood. he ...201323307809
glutamate efflux mediated by corynebacterium glutamicum msccg, escherichia coli mscs, and their derivatives.corynebacterium glutamicum is used in microbial biotechnology for the production of amino acids, in particular glutamate. the mechanism of glutamate excretion, however, is not yet fully understood. recently, evidence was provided that the ncgl1221 gene product from c. glutamicum atcc 13869, a mscs-type mechanosensitive efflux channel, is responsible for glutamate efflux [1]. the major difference of ncgl1221 and the homologous protein msccg of c. glutamicum atcc 13032 from escherichia coli mscs a ...201323313454
increasing l-isoleucine production in corynebacterium glutamicum by overexpressing global regulator lrp and two-component export system brnfe.to increase the l-isoleucine production in corynebacterium glutamicum by overexpressing the global regulator lrp and the two-component export system brnfe.201323331988
increasing the antibacterial activity of gentamicin in combination with extracted polyphosphate from bacillus megaterium.the aim of this research was production of polyphosphate (poly p) and study on its antibacterial effects.201323332009
synthesis, structural elucidation, and biochemical analysis of immunoactive glucuronosyl diacylglycerides of mycobacteria and corynebacteria.glucuronosyl diacylglycerides (glcagroac2) are functionally important glycolipids and membrane anchors for cell wall lipoglycans in the corynebacteria. here we describe the complete synthesis of distinct acyl-isoforms of glcagroac2 bearing both acylation patterns of (r)-tuberculostearic acid (c19:0) and palmitic acid (c16:0) and their mass spectral characterization. collision-induced fragmentation mass spectrometry identified characteristic fragment ions that were used to develop "rules" allowin ...201323343519
genr, an iclr-type regulator, activates and represses the transcription of gen genes involved in 3-hydroxybenzoate and gentisate catabolism in corynebacterium glutamicum.the genes required for 3-hydroxybenzoate and gentisate catabolism in corynebacterium glutamicum are closely clustered in three operons. genr, an iclr-type regulator, can activate the transcription of genkh and gendfm operons in response to 3-hydroxybenzoate and gentisate, and it can repress its own expression. footprinting analyses demonstrated that genr bound to four sites with different affinities. two genr-binding sites (dfmn01 and dfmn02) were found to be located between positions --41 and - ...201323354754
modification of histidine biosynthesis pathway genes and the impact on production of l-histidine in corynebacterium glutamicum.histidine biosynthesis in corynebacterium glutamicum is regulated not only by feedback inhibition by the first enzyme in the pathway, but also by repression control of the synthesis of the histidine enzymes. c. glutamicum histidine genes are located and transcribed in two unlinked loci, hiseg and hisdcb-orf1-orf2-hisha-impa-hisfi. we constructed plasmid pk18hisdptac to replace the native hisd promoter with the tac promoter, and overexpressed phosphoribosyl-atp-pyrophosphohydrolase, encoded by hi ...201323355034
evaluation of the food grade expression systems nice and psip for the production of 2,5-diketo-d-gluconic acid reductase from corynebacterium glutamicum.2,5-diketo-d-gluconic acid reductase (2,5-dkg reductase) catalyses the reduction of 2,5-diketo-d-gluconic acid (2,5-dkg) to 2-keto-l-gulonic acid (2-klg), a direct precursor (lactone) of l-ascorbic acid (vitamin c). this reaction is an essential step in the biocatalytic production of the food supplement vitamin c from d-glucose or d-gluconic acid. as 2,5-dkg reductase is usually produced recombinantly, it is of interest to establish an efficient process for 2,5-dkg reductase production that also ...201323356419
nrdh-redoxin of mycobacterium tuberculosis and corynebacterium glutamicum dimerizes at high protein concentration and exclusively receives electrons from thioredoxin reductase.nrdh-redoxins are small reductases with a high amino acid sequence similarity with glutaredoxins and mycoredoxins but with a thioredoxin-like activity. they function as the electron donor for class ib ribonucleotide reductases, which convert ribonucleotides into deoxyribonucleotides. we solved the x-ray structure of oxidized nrdh-redoxin from corynebacterium glutamicum (cg) at 1.5 å resolution. based on this monomeric structure, we built a homology model of nrdh-redoxin from mycobacterium tuberc ...201323362277
the salicylate 1,2-dioxygenase as a model for a conventional gentisate 1,2-dioxygenase: crystal structures of the g106a mutant and its adducts with gentisate and salicylate.the salicylate 1,2-dioxygenase (sdo) from the bacterium pseudaminobacter salicylatoxidans bn12 is a versatile gentisate 1,2-dioxygenase (gdo) that converts both gentisate (2,5-dihydroxybenzoate) and various monohydroxylated substrates. several variants of this enzyme were rationally designed based on the previously determined enzyme structure and sequence differences between the sdo and the 'conventional' gdo from corynebacterium glutamicum. this was undertaken in order to define the structural ...201323384287
phosphotransferase system-mediated glucose uptake is repressed in phosphoglucoisomerase-deficient corynebacterium glutamicum strains.corynebacterium glutamicum is particularly known for its industrial application in the production of amino acids. amino acid overproduction comes along with a high nadph demand, which is covered mainly by the oxidative part of the pentose phosphate pathway (ppp). in previous studies, the complete redirection of the carbon flux toward the ppp by chromosomal inactivation of the pgi gene, encoding the phosphoglucoisomerase, has been applied for the improvement of c. glutamicum amino acid production ...201323396334
involvement of regulatory interactions among global regulators glxr, sugr, and rama in expression of rama in corynebacterium glutamicum.the central carbon metabolism genes in corynebacterium glutamicum are under the control of a transcriptional regulatory network composed of several global regulators. it is known that the promoter region of rama, encoding one of these regulators, interacts with its gene product, rama, as well as with the two other regulators, glxr and sugr, in vitro and/or in vivo. although rama has been confirmed to repress its own expression, the roles of glxr and sugr in rama expression have remained unclear. ...201323396909
metabolic engineering of corynebacterium glutamicum to produce gdp-l-fucose from glucose and mannose.wild-type corynebacterium glutamicum was metabolically engineered to convert glucose and mannose into guanosine 5'-diphosphate (gdp)-l-fucose, a precursor of fucosyl-oligosaccharides, which are involved in various biological and pathological functions. this was done by introducing the gmd and wcag genes of escherichia coli encoding gdp-d-mannose-4,6-dehydratase and gdp-4-keto-6-deoxy-d-mannose-3,5-epimerase-4-reductase, respectively, which are known as key enzymes in the production of gdp-l-fuco ...201323404100
chromosome segregation impacts on cell growth and division site selection in corynebacterium glutamicum.spatial and temporal regulation of bacterial cell division is imperative for the production of viable offspring. in many rod-shaped bacteria, regulatory systems such as the min system and nucleoid occlusion ensure the high fidelity of midcell divisome positioning. however, regulation of division site selection in bacteria lacking recognizable min and nucleoid occlusion remains less well understood. here, we describe one such rod-shaped organism, corynebacterium glutamicum, which does not always ...201323405112
media optimization of corynebacterium glutamicum for succinate production under oxygen-deprived condition.corynebacterium glutamicum is one of the well-studied industrial strain that is used for the production of nucleotides and amino acids. recently, it has also been studied as a possible producer of organic acids such as succinic acid, based on its ability to produce organic acids under an oxygen deprivation condition. in this study, we conducted the optimization of medium components for improved succinate production from c. glutamicum under an oxygen deprivation condition by plackett-burman desig ...201323412064
conversion of corynebacterium glutamicum from an aerobic respiring to an aerobic fermenting bacterium by inactivation of the respiratory chain.in this study a comparative analysis of three corynebacterium glutamicum atcc 13032 respiratory chain mutants lacking either the cytochrome bd branch (δcydab), or the cytochrome bc1-aa3 branch (δqcr), or both branches was performed. the lack of cytochrome bd oxidase was inhibitory only under conditions of oxygen limitation, whereas the absence of a functional cytochrome bc1-aa3 supercomplex led to decreases in growth rate, biomass yield, respiration and proton-motive force (pmf) and a strongly i ...201323416842
proteome turnover in bacteria: current status for corynebacterium glutamicum and related bacteria.with the advent of high-resolution mass spectrometry together with sophisticated data analysis and interpretation algorithms, determination of protein synthesis and degradation rates (i.e. protein turnover) on a proteome-wide scale by employing stable isotope-labelled amino acids has become feasible. these dynamic data provide a deeper understanding of protein homeostasis and stress response mechanisms in microorganisms than well-established 'steady state' proteomics approaches. in this article, ...201323425033
systems metabolic engineering of xylose-utilizing corynebacterium glutamicum for production of 1,5-diaminopentane.the sustainable production of industrial platform chemicals is one of the great challenges facing the biotechnology field. ideally, fermentation feedstocks would rather rely on industrial waste streams than on food-based raw materials. corynebacterium glutamicum was metabolically engineered to produce the bio-nylon precursor 1,5-diaminopentane from the hemicellulose sugar xylose. comparison of a basic diaminopentane producer strain on xylose and glucose feedstocks revealed a 30% reduction in dia ...201323447448
interaction between dahp synthase and chorismate mutase endows new regulation on dahp synthase activity in corynebacterium glutamicum.previous research on corynebacterium glutamicum revealed that 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (dscg, formerly ds2098) interacts with chorismate mutase (cmcg, formerly cm0819). in this study, we investigated the interaction by means of structure-guided mutation and enzymatic assays. our results show that the interaction imparted a new mechanism for regulation of dahp activity: in the absence of cmcg, dscg activity was not regulated by prephenate, whereas in the presence of cm ...201323467831
kinetic and thermodynamic characterization of lysine production process in brevibacterium lactofermentum.detailed kinetic and thermodynamic parameters for lysine production from brevibacterium lactofermentum are investigated for the first time in this study. production of the essential amino acid, l-lysine, by b. lactofermentum was assessed in a flask and a continuously stirred tank fermentor (22 l). maximum lysine production was achieved after 40 h of growth and at 35 °c. the effect of different nitrogen sources such as nh(4)no(3), (nh(4))(2)so(4), (nh(4))(2)hpo(4), corn steep liquor, nano(3), and ...201323475286
enhanced production of l-phenylalanine in corynebacterium glutamicum due to the introduction of escherichia coli wild-type gene aroh.metabolic engineering is a powerful tool which has been widely used for producing valuable products. for improving l-phenylalanine (l-phe) accumulation in corynebacterium glutamicum, we have investigated the target genes involved in the biosynthetic pathways. the genes involved in the biosynthesis of l-phe were found to be strictly regulated genes by feedback inhibition. as a result, overexpression of the native wild-type genes arof, arog or phea resulted in a slight increase of l-phe. in contra ...201323526182
maltose uptake by the novel abc transport system musefgk2i causes increased expression of ptsg in corynebacterium glutamicum.the gram-positive corynebacterium glutamicum efficiently metabolizes maltose by a pathway involving maltodextrin and glucose formation by 4-α-glucanotransferase, glucose phosphorylation by glucose kinases, and maltodextrin degradation via maltodextrin phosphorylase and α-phosphoglucomutase. however, maltose uptake in c. glutamicum has not been investigated. interestingly, the presence of maltose in the medium causes increased expression of ptsg in c. glutamicum by an unknown mechanism, although ...201323543710
sample preparation, crystallization and structure solution of hisc from mycobacterium tuberculosis.histidinolphosphate aminotransferase (hisc; rv1600) from mycobacterium tuberculosis was overexpressed in m. smegmatis and purified to homogeneity using nickel-nitrilotriacetic acid metal-affinity and gel-filtration chromatography. diffraction-quality crystals suitable for x-ray analysis were grown by the hanging-drop vapour-diffusion technique using 30% polyethylene glycol monomethyl ether 2000 as the precipitant. the crystals belonged to the hexagonal space group p3221, with an unusual high sol ...201323545656
impact of carbon source and variable nitrogen conditions on bacterial biosynthesis of polyhydroxyalkanoates: evidence of an atypical metabolism in bacillus megaterium dsm 509.twenty bacterial strains were examined on their ability to produce polyhydroxyalkanoates (pha) from different carbon sources under rich and depleted nitrogen conditions. preliminary experiments with glucose as sole carbon source allowed to select pha producing bacteria using ftir spectroscopy. they were further tested with eight additional carbon substrates including organic, fatty acids or sugars. pha content and monomer composition of four chosen strains (pseudomonas putida mt-2, bacillus mega ...201323548274
crude glycerol-based production of amino acids and putrescine by corynebacterium glutamicum.corynebacterium glutamicum possesses genes for glycerol kinase and glycerol-3-phosphate dehydrogenase that were shown to support slow growth with glycerol only when overexpressed from a plasmid. pure glycerol and crude glycerol from biodiesel factories were tested for growth of recombinant strains expressing glpf, glpk and glpd from escherichia coli. some, but not all crude glycerol lots served as good carbon sources. although the inhibitory compound(s) present in these crude glycerol lots remai ...201323562176
crystal and solution studies reveal that the transcriptional regulator acnr of corynebacterium glutamicum is regulated by citrate-mg2+ binding to a non-canonical pocket.corynebacterium glutamicum is an important industrial bacterium as well as a model organism for the order corynebacteriales, whose citric acid cycle occupies a central position in energy and precursor supply. expression of aconitase, which isomerizes citrate into isocitrate, is controlled by several transcriptional regulators, including the dimeric aconitase repressor acnr, assigned by sequence identity to the tetr family. we report the structures of acnr in two crystal forms together with ligan ...201323589369
oxygen supply in disposable shake-flasks: prediction of oxygen transfer rate, oxygen saturation and maximum cell concentration during aerobic growth.dissolved oxygen plays an essential role in aerobic cultivation especially due to its low solubility. under unfavorable conditions of mixing and vessel geometry it can become limiting. this, however, is difficult to predict and thus the right choice for an optimal experimental set-up is challenging. to overcome this, we developed a method which allows a robust prediction of the dissolved oxygen concentration during aerobic growth. this integrates newly established mathematical correlations for t ...201323592306
engineering of acetate recycling and citrate synthase to improve aerobic succinate production in corynebacterium glutamicum.corynebacterium glutamicum lacking the succinate dehydrogenase complex can produce succinate aerobically with acetate representing the major byproduct. efforts to increase succinate production involved deletion of acetate formation pathways and overexpression of anaplerotic pathways, but acetate formation could not be completely eliminated. to address this issue, we constructed a pathway for recycling wasted carbon in succinate-producing c. glutamicum. the acetyl-coa synthetase from bacillus sub ...201323593275
development of a secretion system for the production of heterologous proteins in corynebacterium glutamicum using the porin b signal peptide.corynebacterium glutamicum is one of the useful hosts for the secretory production of heterologous proteins because of intrinsic attributes such as the presence of few endogenous proteins and proteases in culture medium. here, we report the development of a new secretory system for the production of heterologous proteins by using the porin b (porb) signal peptide in c. glutamicum. we examined two different endoxylanases and an antibody fragment (scfv) as model proteins for secretory production. ...201323597779
regulatory interaction of the corynebacterium glutamicum whc genes in oxidative stress responses.in this study, we analyzed the regulatory interaction of the corynebacterium glutamicum whc genes that play roles in oxidative stress responses. we found that whce and whca transcription was minimal in the whcb-deleted mutant (δwhcb). however, whcb and whca transcription increased in the δwhce mutant during the log phase, whereas their transcription decreased during the stationary phase. in addition, cells carrying the p180-whcb vector, which showed retarded growth due to uncontrolled whcb overe ...201323608553
recovery of high-purity metallic pd from pd(ii)-sorbed biosorbents by incineration.this work reports a direct way to recover metallic palladium with high purity from pd(ii)-sorbed polyethylenimine-modified corynebacterium glutamicum biosorbent using a combined method of biosorption and incineration. this study is focused on the incineration part which affects the purity of recovered pd. the incineration temperature and the amount of pd loaded on the biosorbent were considered as major factors in the incineration process, and their effects were examined. the results showed that ...201323611701
physiological roles of mycothiol in detoxification and tolerance to multiple poisonous chemicals in corynebacterium glutamicum.mycothiol (msh) plays important roles in maintaining cytosolic redox homeostasis and in adapting to reactive oxygen species in the high-(g + c)-content gram-positive actinobacteria. however, its physiological roles are ill defined compared to glutathione, the functional analog of msh in gram-negative bacteria and most eukaryotes. in this research, we explored the impact of intracellular msh on cellular physiology by using msh-deficient mutants in the model organism corynebacterium glutamicum. we ...201323615850
histidine biosynthesis, its regulation and biotechnological application in corynebacterium glutamicum.l-histidine biosynthesis is an ancient metabolic pathway present in bacteria, archaea, lower eukaryotes, and plants. for decades l-histidine biosynthesis has been studied mainly in escherichia coli and salmonella typhimurium, revealing fundamental regulatory processes in bacteria. furthermore, in the last 15 years this pathway has been also investigated intensively in the industrial amino acid-producing bacterium corynebacterium glutamicum, revealing similarities to e. coli and s. typhimurium, a ...201423617600
oxyr acts as a transcriptional repressor of hydrogen peroxide-inducible antioxidant genes in corynebacterium glutamicum r.oxyr, a lysr-type transcriptional regulator, has been established as a redox-responsive activator of antioxidant genes in bacteria. this study shows that oxyr acts as a transcriptional repressor of kata, dps, ftn and cyda in corynebacterium glutamicum r. kata encodes h2o2-detoxifing enzyme catalase, dps and ftn are implicated in iron homeostasis and cyda encodes a subunit of cytochrome bd oxidase. quantitative rt-pcr analyses revealed that expression of kata and dps, but not of ftn and cyda, was ...201323621709
proteome response of corynebacterium glutamicum to high concentration of industrially relevant c₄ and c₅ dicarboxylic acids.more than fifty years of industrial and scientific developments on the amino acid-producer strain corynebacterium glutamicum has generated an extremely huge knowledge highly applicable to the development of new products. despite the production of dicarboxylic acids has already been engineered in c. glutamicum, the effect caused by these acids at competitive industrial levels has not yet been described. thus, aspartic, fumaric, itaconic, malic and succinic acids have been tested on the growth of ...201323624027
recombineering in corynebacterium glutamicum combined with optical nanosensors: a general strategy for fast producer strain generation.recombineering in bacteria is a powerful technique for genome reconstruction, but until now, it was not generally applicable for development of small-molecule producers because of the inconspicuous phenotype of most compounds of biotechnological relevance. here, we establish recombineering for corynebacterium glutamicum using rect of prophage rac and combine this with our recently developed nanosensor technology, which enables the detection and isolation of productive mutants at the single-cell ...201323630315
isolation of fully synthetic promoters for high-level gene expression in corynebacterium glutamicum.corynebacterium glutamicum is an important industrial organism that is widely used in the production of amino acids, nucleotides and vitamins. to extend its product spectrum and improve productivity, c. glutamicum needs to undergo further engineering, including the development of applicable promoter system. here, we isolated new promoters from the fully synthetic promoter library consisting of 70-bp random sequences in c. glutamicum. using green fluorescent protein (gfp) as a reporter, highly fl ...201323633298
cord factor (trehalose 6,6'-dimycolate) forms fully stable and non-permeable lipid bilayers required for a functional outer membrane.cord factor (trehalose 6,6'-dimycolate, tdm) is the major lipid in the outer membrane of corynebacteria and mycobacteria. although its role is well recognized in the immune response phenomena, its membrane biophysical properties remained largely unexplored and tdm has often been described as a detergent. we purified the main components of the outer membrane from corynebacterium glutamicum and analyzed their membrane forming properties. in mixture with endogenous cardiolipin, but not alone, the s ...201323643889
l-glutamate secretion by the n-terminal domain of the corynebacterium glutamicum ncgl1221 mechanosensitive channel.the corynebacterium glutamicum ncgl1221 mechanosensitive channel mediates l-glutamate secretion by sensing changes in membrane tension caused by treatments such as biotin limitation and penicillin. the ncgl1221 protein has an n-terminal domain (1-286 a.a.) homologous to the escherichia coli mscs and a long c-terminal domain (287-533 a.a.) of unknown function. in order to investigate the role of the c-terminal domain in l-glutamate secretion, we constructed a series of c-terminally truncated muta ...201323649271
transcriptional analysis of the groes-groel1, groel2, and dnak genes in corynebacterium glutamicum: characterization of heat shock-induced promoters. 201323661686
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