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development of a new platform for secretory production of recombinant proteins in corynebacterium glutamicum.corynebacterium glutamicum, which has been for long an industrial producer of various l-amino acids, nucleic acids, and vitamins, is now also regarded as a potential host for the secretory production of recombinant proteins. to harness its potential as an industrial platform for recombinant protein production, the development of an efficient secretion system is necessary. particularly, regarding protein production in large-scale bioreactors, it would be appropriate to develop a secretory express ...201626134574
structure and function of amtr in mycobacterium smegmatis: implications for post-transcriptional regulation of urea metabolism through a small antisense rna.soil-dwelling bacteria of the phylum actinomycetes generally harbor either glnr or amtr as a global regulator of nitrogen metabolism. mycobacterium smegmatis harbors both of these canonical regulators; glnr regulates the expression of key genes involved in nitrogen metabolism, while the function and signal transduction pathway of amtr in m. smegmatis remains largely unknown. here, we report the structure and function of the m. smegmatis amtr and describe the role of amtr in the regulation of nit ...201627640309
corrigendum: cell division in corynebacterineae.[this corrects the article on p. 132 in vol. 5, pmid: 24782835.].201627990143
structure-function profile of mmpl3, the essential mycolic acid transporter from mycobacterium tuberculosis.the mmpl family of proteins translocates complex (glyco)lipids and siderophores across the cell envelope of mycobacteria and closely related corynebacteriaceae and plays important roles in the biogenesis of the outer membrane of these organisms. despite their significance in the physiology and virulence of mycobacterium tuberculosis, and from the perspective of developing novel antituberculosis agents, little is known about their structure and mechanism of translocation. in this study, the essen ...201627737557
assembling of the mycobacterium tuberculosis cell wall core.the unique cell wall of mycobacteria is essential to their viability and the target of many clinically used anti-tuberculosis drugs and inhibitors under development. despite intensive efforts to identify the ligase(s) responsible for the covalent attachment of the two major heteropolysaccharides of the mycobacterial cell wall, arabinogalactan (ag) and peptidoglycan (pg), the enzyme or enzymes responsible have remained elusive. we here report on the identification of the two enzymes of mycobacter ...201627417139
3-methyl-1-butanol biosynthesis in an engineered corynebacterium glutamicum.biofuel offers a promising solution to the adverse environmental problems and depletion in reserves of fossil fuels. higher alcohols including 3-methyl-1-butanol were paid much more attention as fuel substitute in recent years, due to its similar properties to gasoline. in the present work, 3-methyl-1-butanol production in engineered corynebacterium glutamicum was studied. α-ketoisovalerate decarboxylase gene (kivd) from lactococcus lactis combined with alcohol dehydrogenase gene (adh2, adha, an ...201626961908
structural insight into dihydrodipicolinate reductase from corybebacterium glutamicum for lysine biosynthesis.dihydrodipicolinate reductase is an enzyme that converts dihydrodipicolinate to tetrahydrodipicolinate using an nad(p)h cofactor in l-lysine biosynthesis. to increase the understanding of the molecular mechanisms of lysine biosynthesis, we determined the crystal structure of dihydrodipicolinate reductase from corynebacterium glutamicum (cgdapb). cgdapb functions as a tetramer, and each protomer is composed of two domains, an nterminal domain and a c-terminal domain. the n-terminal domain mainly ...201626502738
structural basis for cytokinin production by log from corynebacterium glutamicum."lonely guy" (log) has been identified as a cytokinin-producing enzyme in plants and plant-interacting fungi. the gene product of cg2612 from the soil-dwelling bacterium corynebacterium glutamicum was annotated as an ldc. however, the facts that c. glutamicum lacks an ldc and cg2612 has high amino acid similarity with log proteins suggest that cg2612 is possibly an log protein. to investigate the function of cg2612, we determined its crystal structure at a resolution of 2.3 å. cg2612 functions a ...201627507425
production of orotic acid by a klura3δ mutant of kluyveromyces lactis.we demonstrated that a klura3δ, mutant of the yeast kluyveromyces lactis is able to produce and secrete into the growth medium considerable amounts of orotic acid. using yeast extract-peptone-glucose (ypd) based media we optimized production conditions in flask and bioreactor cultures. with cells grown in ypd 5% glucose medium, the best production in flask was obtained with a 1:12.5 ratio for flask: culture volume, 180 rpm, 28°c and 200 mm mops for ph stabilization at neutral values (initial cul ...201626707627
impact of lytr-cpsa-psr proteins on cell wall biosynthesis in corynebacterium glutamicum.proteins of the lcp (lytr, cpsa, psr) family have been shown to inherit important roles in bacterial cell wall biosynthesis. however, their exact function in the formation of the complex cell wall structures of the corynebacteriales, including the prominent pathogens mycobacterium tuberculosis and corynebacterium diphtheriae, remains unclear. here, we analyzed the role of the lcp proteins lcpa and lcpb of corynebacterium glutamicum, both of which localize at regions of nascent cell wall biosynth ...201627551018
caenorhabditis elegans star formation and negative chemotaxis induced by infection with corynebacteria.caenorhabditis elegans is one of the major model systems in biology based on advantageous properties such as short life span, transparency, genetic tractability and ease of culture using an escherichia coli diet. in its natural habitat, compost and rotting plant material, this nematode lives on bacteria. however, c. elegans is a predator of bacteria, but can also be infected by nematopathogenic coryneform bacteria such microbacterium and leucobacter species, which display intriguing and diverse ...201626490043
expression of recombinant protein using corynebacterium glutamicum: progress, challenges and applications.corynebacterium glutamicum (c. glutamicum) is a highly promising alternative prokaryotic host for recombinant protein expression, as it possesses several significant advantages over escherichia coli (e. coli), the currently leading bacterial protein expression system. during the past decades, several experimental techniques and vector components for genetic manipulation of c. glutamicum have been developed and validated, including strong promoters for tightly regulating target gene expression, v ...201625714007
branched-chain amino acids.branched-chain amino acids (bcaas), viz., l-isoleucine, l-leucine, and l-valine, are essential amino acids that cannot be synthesized in higher organisms and are important nutrition for humans as well as livestock. they are also valued as synthetic intermediates for pharmaceuticals. therefore, the demand for bcaas in the feed and pharmaceutical industries is increasing continuously. traditional industrial fermentative production of bcaas was performed using microorganisms isolated by random muta ...201627872960
biotechnological production of aromatic compounds of the extended shikimate pathway from renewable biomass.aromatic chemicals that contain an unsaturated ring with alternating double and single bonds find numerous applications in a wide range of industries, e.g. paper and dye manufacture, as fuel additives, electrical insulation, resins, pharmaceuticals, agrochemicals, in food, feed and cosmetics. their chemical production is based on petroleum (btx; benzene, toluene, and xylene), but they can also be obtained from plants by extraction. due to petroleum depletion, health compliance, or environmental ...201627871872
exporters for production of amino acids and other small molecules.microbes are talented catalysts to synthesize valuable small molecules in their cytosol. however, to make full use of their skills - and that of metabolic engineers - the export of intracellularly synthesized molecules to the culture medium has to be considered. this step is as essential as is each step for the synthesis of the favorite molecule of the metabolic engineer, but is frequently not taken into account. to export small molecules via the microbial cell envelope, a range of different typ ...201627832297
lysine fermentation: history and genome breeding.lysine fermentation by corynebacterium glutamicum was developed in 1958 by kyowa hakko kogyo co. ltd. (current kyowa hakko bio co. ltd.) and is the second oldest amino acid fermentation process after glutamate fermentation. the fundamental mechanism of lysine production, discovered in the early stages of the process's history, gave birth to the concept known as "metabolic regulatory fermentation," which is now widely applied to metabolite production. after the development of rational metabolic e ...201627832296
increased production of food-grade d-tagatose from d-galactose by permeabilized and immobilized cells of corynebacterium glutamicum, a gras host, expressing d-galactose isomerase from geobacillus thermodenitrificans.the generally recognized as safe microorganism corynebacterium glutamicum expressing geobacillus thermodenitrificans d-galactose isomerase (d-gai) was an efficient host for the production of d-tagatose, a functional sweetener. the d-tagatose production at 500 g/l d-galactose by the host was 1.4-fold higher than that by escherichia coli expressing d-gai. the d-tagatose-producing activity of permeabilized c. glutamicum (pcg) cells treated with 1% (w/v) triton x-100 was 2.1-fold higher than that of ...201627734668
development of a genetically engineered escherichia coli strain for plasmid transformation in corynebacterium glutamicum.gene disruption and replacement in corynebacterium glutamicum is dependent upon a high transformation efficiency. the cglir-cgiir restriction system is a major barrier to introduction of foreign dna into corynebacterium glutamicum cells. to improve the transformation efficiency of c. glutamicum, the cglim gene encoding methyltransferase in the cglir-cgliir-cglim restriction-modification system of c. glutamicum atcc 13032 was chromosomally integrated and expressed in escherichia coli, resulting i ...201627793586
a new metabolic route for the production of gamma-aminobutyric acid by corynebacterium glutamicum from glucose.gamma-aminobutyric acid (gaba), a non-protein amino acid widespread in nature, is a component of pharmaceuticals, foods, and the biodegradable plastic polyamide 4. corynebacterium glutamicum shows great potential for the production of gaba from glucose. gaba added to the growth medium hardly affected growth of c. glutamicum, since a half-inhibitory concentration of 1.1 m gaba was determined. as alternative to gaba production by glutamate decarboxylation, a new route for the production of gaba vi ...201627289384
overexpression of methionine adenosyltransferase in corynebacterium glutamicum for production of s-adenosyl-l-methionine.two genes encoding methionine adenosyltransferase, sam2 from saccharomyces cerevisiae and metk from corynebacterium glutamicum, were individually cloned into pdxw-8, the shuttle vector between escherichia coli and c. glutamicum, and overexpressed in e. coli dh5α and c. glutamicum atcc13032. in dh5α, both genes were overexpressed and their protein products showed the activity of methionine adenosyltransferase. in atcc13032, metk was overexpressed, its product metk showed the enzyme activity and c ...201626238196
stereospecificity of corynebacterium glutamicum 2,3-butanediol dehydrogenase and implications for the stereochemical purity of bioproduced 2,3-butanediol.the stereochemistry of 2,3-butanediol (2,3-bd) synthesis in microbial fermentations is important for many applications. in this work, we showed that corynebacterium glutamicum endowed with the lactococcus lactis genes encoding α-acetolactate synthase and decarboxylase activities produced meso-2,3-bd as the major end product, meaning that (r)-acetoin is a substrate for endogenous 2,3-butanediol dehydrogenase (bdh) activity. this is curious in view of the reported absolute stereospecificity of c. ...201627687994
the topology of the l-arginine exporter argo conforms to an nin-cout configuration in escherichia coli: requirement for the cytoplasmic n-terminal domain, functional helical interactions, and an aspartate pair for argo function.argo and lyse are members of the lyse family of exporter proteins and ordinarily mediate the export of l-arginine (arg) in escherichia coli and l-lysine (lys) and arg in corynebacterium glutamicum, respectively. under certain conditions, argo also mediates lys export. to delineate the arrangement of argo in the cytoplasmic membrane of e. coli, we have employed a combination of cysteine accessibility in situ, alkaline phosphatase fusion reporters, and protein modeling to arrive at a topological m ...201627645388
time-resolved, single-cell analysis of induced and programmed cell death via non-invasive propidium iodide and counterstain perfusion.conventional propidium iodide (pi) staining requires the execution of multiple steps prior to analysis, potentially affecting assay results as well as cell vitality. in this study, this multistep analysis method has been transformed into a single-step, non-toxic, real-time method via live-cell imaging during perfusion with 0.1 μm pi inside a microfluidic cultivation device. dynamic pi staining was an effective live/dead analytical tool and demonstrated consistent results for single-cell death in ...201627580964
high-yield anaerobic succinate production by strategically regulating multiple metabolic pathways based on stoichiometric maximum in escherichia coli.succinate has been identified by the u.s. department of energy as one of the top 12 building block chemicals, which can be used as a specialty chemical in the agricultural, food, and pharmaceutical industries. escherichia coli are now one of the most important succinate producing candidates. however, the stoichiometric maximum succinate yield under anaerobic conditions through the reductive branch of the tca cycle is restricted by nadh supply in e. coli.201627520031
the flexible feedstock concept in industrial biotechnology: metabolic engineering of escherichia coli, corynebacterium glutamicum, pseudomonas, bacillus and yeast strains for access to alternative carbon sources.most biotechnological processes are based on glucose that is either present in molasses or generated from starch by enzymatic hydrolysis. at the very high, million-ton scale production volumes, for instance for fermentative production of the biofuel ethanol or of commodity chemicals such as organic acids and amino acids, competing uses of carbon sources e.g. in human and animal nutrition have to be taken into account. thus, the biotechnological production hosts e. coli, c. glutamicum, pseudomona ...201627491712
d-allulose production from d-fructose by permeabilized recombinant cells of corynebacterium glutamicum cells expressing d-allulose 3-epimerase flavonifractor plautii.a d-allulose 3-epimerase from flavonifractor plautii was cloned and expressed in escherichia coli and corynebacterium glutamicum. the maximum activity of the enzyme purified from recombinant e. coli cells was observed at ph 7.0, 65°c, and 1 mm co2+ with a half-life of 40 min at 65°c, km of 162 mm, and kcat of 25280 1/s. for increased d-allulose production, recombinant c. glutamicum cells were permeabilized via combined treatments with 20 mg/l penicillin and 10% (v/v) toluene. under optimized con ...201627467527
progress in the microbial production of s-adenosyl-l-methionine.s-adenosyl-l-methionine (sam), which exists in all living organisms, serves as an activated group donor in a range of metabolic reactions, including trans-methylation, trans-sulfuration and trans-propylamine. compared with its chemical synthesis and enzyme catalysis production, the microbial production of sam is feasible for industrial applications. the current clinical demand for sam is constantly increasing. therefore, vast interest exists in engineering the sam metabolism in cells for increas ...201627465853
engineering a glycerol utilization pathway in corynebacterium glutamicum for succinate production under o2 deprivation.to explore the glycerol utilization pathway in corynebacterium glutamicum for succinate production under o2 deprivation.201627395064
enhanced succinate production from glycerol by engineered escherichia coli strains.in this study, an engineered strain escherichia coli mlb (ldha(-)pflb(-)) was constructed for production of succinate from glycerol. the succinate yield was 0.37mol/mol in anaerobic culture, however, the growth and glycerol consumption rates were very slow, resulting in a low succinate level. two-stage fermentation was performed in flasks, and the succinate yield reached 0.93mol/mol, but the succinate titer was still low. hence, overexpression of malate dehydrogenase, malic enzyme, phosphoenolpy ...201627371794
roles of export genes cgma and lyse for the production of l-arginine and l-citrulline by corynebacterium glutamicum.l-arginine is a semi-essential amino acid with application in cosmetic, pharmaceutical, and food industries. metabolic engineering strategies have been applied for overproduction of l-arginine by corynebacterium glutamicum. lyse was the only known l-arginine exporter of this bacterium. however, an l-arginine-producing strain carrying a deletion of lyse still accumulated about 10 mm l-arginine in the growth medium. overexpression of the putative putrescine and cadaverine export permease gene cgma ...201627350619
three-steps in one-pot: whole-cell biocatalytic synthesis of enantiopure (+)- and (-)-pinoresinol via kinetic resolution.pinoresinol is a high-value plant-derived lignan with multiple health supporting effects. enantiomerically pure pinoresinol can be isolated from natural sources, but with low efficiency. most chemical and biocatalytic approaches that have been described for the synthesis of pinoresinol furnish the racemic mixture. in this study we devised a three-step biocatalytic cascade for the production of enantiomerically pure pinoresinol from the cheap compound eugenol. two consecutive oxidations of eugeno ...201627160378
updates on industrial production of amino acids using corynebacterium glutamicum.l-amino acids find various applications in biotechnology. l-glutamic acid and its salts are used as flavor enhancers. other l-amino acids are used as food or feed additives, in parenteral nutrition or as building blocks for the chemical and pharmaceutical industries. l-amino acids are synthesized from precursors of central carbon metabolism. based on the knowledge of the biochemical pathways microbial fermentation processes of food, feed and pharma amino acids have been developed. production str ...201627116971
properties of cassava starch modified by amylomaltase from corynebacterium glutamicum.amylomaltase (α-1,4-glucanotransferase, am; ec 2.4.1.25) from corynebacterium glutamicum expressed in escherichia coli was used to prepare the enzyme-modified cassava starch for food application. about 5% to 15% (w/v) of cassava starch slurries were incubated with 1, 3, or 5 units of amylomaltase/g starch. apparent amylose, amylopectin chain length distribution, thermal properties, freeze-thaw stability, thermo-reversibility, and gel strength of the obtained modified starches were measured. the ...201627105125
influence of nanomechanical stress induced by zno nanoparticles of different shapes on the viability of cells.there is growing interest in nanostructures interacting with living organisms. however, there are still no general rules for the design of biocompatible nanodevices. here, we present a step towards understanding the interactions between nanostructures and living cells. we study the influence of nanomechanical stress induced by zinc oxide (zno) nanostructures of different shapes on the viability of both prokaryotic (gram-negative bacteria: escherichia coli and enterobacter aerogenes, and gram-pos ...201627074722
dihydroxyacetone production in an engineered escherichia coli through expression of corynebacterium glutamicum dihydroxyacetone phosphate dephosphorylase.dihydroxyacetone (dha) has several industrial applications such as a tanning agent in tanning lotions in the cosmetic industry; its production via microbial fermentation would present a more sustainable option for the future. here we genetically engineered escherichia coli (e. coli) for dha production from glucose. deletion of e. coli triose phosphate isomerase (tpia) gene was carried out to accumulate dihydroxyacetone phosphate (dhap), for use as the main intermediate or precursor for dha produ ...201626992791
production of succinic acid by metabolically engineered microorganisms.succinic acid (sa) has been recognized as one of the most important bio-based building block chemicals due to its numerous potential applications. for the economical bio-based production of sa, extensive research works have been performed on developing microbial strains by metabolic engineering as well as fermentation and downstream processes. here we review metabolic engineering strategies applied for bio-based production of sa using representative microorganisms, including saccharomyces cerevi ...201626990278
pathway construction and metabolic engineering for fermentative production of ectoine in escherichia coli.ectoine is a protective agent and stabilizer whose synthesis pathway exclusively exists in select moderate halophiles. a novel established process called "bacterial milking" efficiently synthesized ectoine in moderate halophiles, however, this method places high demands on equipment and is cost prohibitive. in this study, we constructed an ectoine producing strain by introducing the ectoine synthesis pathway into escherichia coli and improved its production capacity. firstly, the ectabc gene clu ...201626969253
a new strategy for production of 5-aminolevulinic acid in recombinant corynebacterium glutamicum with high yield.5-aminolevulinic acid (ala), a nonprotein amino acid involved in tetrapyrrole synthesis, has been widely applied in agriculture, medicine, and food production. many engineered metabolic pathways have been constructed; however, the production yields are still low. in this study, several 5-aminolevulinic acid synthases (alass) from different sources were evaluated and compared with respect to their ala production capacities in an engineered corynebacterium glutamicum cgs1 strain that can accumulat ...201626921424
microfluidic screening of electric fields for electroporation.electroporation is commonly used to deliver molecules such as drugs, proteins, and/or dna into cells, but the mechanism remains poorly understood. in this work a rapid microfluidic assay was developed to determine the critical electric field threshold required for inducing bacterial electroporation. the microfluidic device was designed to have a bilaterally converging channel to amplify the electric field to magnitudes sufficient to induce electroporation. the bacterial cells are introduced into ...201626893024
strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression.with the availability of the whole genome sequence of escherichia coli or corynebacterium glutamicum, strategies for directed dna manipulation have developed rapidly. dna manipulation plays an important role in understanding the function of genes and in constructing novel engineering bacteria according to requirement. dna manipulation involves modifying the autologous genes and expressing the heterogenous genes. two alternative approaches, using electroporation linear dna or recombinant suicide ...201626834010
web application for genetic modification flux with database to estimate metabolic fluxes of genetic mutants.computational analysis of metabolic fluxes is essential in understanding the structure and function of a metabolic network and in rationally designing genetically modified mutants for an engineering purpose. we had presented the genetic modification flux (gmf) that predicts the flux distribution of a broad range of genetically modified mutants. to enhance the feasibility and usability of gmf, we have developed a web application with a metabolic network database to predict a flux distribution of ...201626777238
in vitro functional characterization of the na+/h+ antiporters in corynebacterium glutamicum.corynebacterium glutamicum, typically used as industrial workhorse for amino acid production, is a moderately salt-alkali-tolerant microorganism with optimal growth at ph 7-9. however, little is known about the mechanisms of salt-alkali tolerance in c. glutamicum. here, the catalytic capacity of three putative na(+)/h(+) antiporters from c. glutamicum (designated as cg-mrp1, cg-mrp2 and cg-nhap) were characterized in an antiporter-deficient escherichia coli knabc strain. only cg-mrp1 was able to ...201626667218
enhanced production of 3-hydroxypropionic acid from glucose via malonyl-coa pathway by engineered escherichia coli.in this study, production of 3-hp via malonyl-coa was investigated by using metabolically engineered escherichia coli carrying heterogeneous acetyl-coa carboxylase (acc) from corynebacterium glutamicum and codon-optimized malonyl-coa reductase (mcr) from chloroflexus aurantiacus. three engineered e. coli strains with different host-vector systems were constructed and investigated. the results indicated that the combination of e. coli bl21(de3) and pet28a was the most efficient host-vector system ...201626606325
fudc, a protein primarily responsible for furfural detoxification in corynebacterium glutamicum.lignocellulosic hydrolysates contain compounds that inhibit microbial growth and fermentation, thereby decreasing the productivity of biofuel and biochemical production. in particular, the heterocyclic aldehyde furfural is one of the most toxic compounds found in these hydrolysates. we previously demonstrated that corynebacterium glutamicum converts furfural into the less toxic compounds furfuryl alcohol and 2-furoic acid. to date, however, the genes involved in these oxidation and reduction rea ...201626541332
the impact of the c-terminal domain on the gating properties of msccg from corynebacterium glutamicum.the mechanosensitive (ms) channel msccg from the soil bacterium corynebacterium glutamicum functions as a major glutamate exporter. msccg belongs to a subfamily of the bacterial mscs-like channels, which play an important role in osmoregulation. to understand the structural and functional features of msccg, we investigated the role of the carboxyl-terminal domain, whose relevance for the channel gating has been unknown. the chimeric channel mscs-(c-msccg), which is a fusion protein between the c ...201626494188
production of protocatechuic acid by corynebacterium glutamicum expressing chorismate-pyruvate lyase from escherichia coli.protocatechuic acid (3,4-dihydroxybenzoic acid; pca) serves as a building block for polymers and pharmaceuticals. in this study, the biosynthetic pathway for pca from glucose was engineered in corynebacterium glutamicum. the pathway to pca-employed elements of the chorismate pathway by using chorismate-pyruvate lyase (cpl) and 4-hydroxybenzoate hydroxylase (4-hba hydroxylase). as c. glutamicum has the potential to synthesize the aromatic amino acid intermediate chorismate and possesses 4-hba hyd ...201626392137
use of a sec signal peptide library from bacillus subtilis for the optimization of cutinase secretion in corynebacterium glutamicum.technical bulk enzymes represent a huge market, and the extracellular production of such enzymes is favorable due to lowered cost for product recovery. protein secretion can be achieved via general secretion (sec) pathway. specific sequences, signal peptides (sps), are necessary to direct the target protein into the translocation machinery. for example, >150 sec-specific sps have been identified for bacillus subtilis alone. as the best sp for a target protein of choice cannot be predicted a prio ...201627927208
engineering corynebacterium glutamicum for fast production of l-lysine and l-pipecolic acid.the gram-positive corynebacterium glutamicum is widely used for fermentative production of amino acids. the world production of l-lysine has surpassed 2 million tons per year. glucose uptake and phosphorylation by c. glutamicum mainly occur by the phosphotransferase system (pts) and to lesser extent by inositol permeases and glucokinases. heterologous expression of the genes for the high-affinity glucose permease from streptomyces coelicolor and bacillus subtilis glucokinase fully compensated fo ...201627345060
identification of d-amino acid dehydrogenase as an upstream regulator of the autoinduction of a putative acyltransferase in corynebacterium glutamicum.expression of a putative acyltransferase encoded by ncgl- 0350 of corynebacterium glutamicum is induced by cell-free culture fluids obtained from stationary-phase growth of both c. glutamicum and pseudomonas aeruginosa, providing evidence for interspecies communication. here, we further confirmed that such communication occurs by showing that acyltransferase expression is induced by culture fluid obtained from diverse gram-negative and -positive bacterial strains, including escherichia coli, sal ...201627225460
diverse effects of a biosurfactant from rhodococcus ruber iegm 231 on the adhesion of resting and growing bacteria to polystyrene.this study evaluated the effects of a trehalolipid biosurfactant produced by rhodococcus ruber iegm 231 on the bacterial adhesion and biofilm formation on the surface of polystyrene microplates. the adhesion of gram-positive (arthrobacter simplex, bacillus subtilis, brevibacterium linens, corynebacterium glutamicum, micrococcus luteus) and gram-negative (escherichia coli, pseudomonas fluorescencens) bacteria correlated differently with the cell hydrophobicity and surface charge. in particular, e ...201626888203
production of the marine carotenoid astaxanthin by metabolically engineered corynebacterium glutamicum.astaxanthin, a red c40 carotenoid, is one of the most abundant marine carotenoids. it is currently used as a food and feed additive in a hundred-ton scale and is furthermore an attractive component for pharmaceutical and cosmetic applications with antioxidant activities. corynebacterium glutamicum, which naturally synthesizes the yellow c50 carotenoid decaprenoxanthin, is an industrially relevant microorganism used in the million-ton amino acid production. in this work, engineering of a genome-r ...201627376307
engineering of corynebacterium glutamicum for xylitol production from lignocellulosic pentose sugars.xylitol is a non-fermentable sugar alcohol used as sweetener. corynebacterium glutamicum atcc13032 was metabolically engineered for xylitol production from the lignocellulosic pentose sugars xylose and arabinose. direct conversion of xylose to xylitol was achieved through the heterologous expression of nad(p)h-dependent xylose reductase (xr) gene from rhodotorula mucilaginosa. xylitol synthesis from arabinose was attained through polycistronic expression of l-arabinose isomerase (araa), d-psicos ...201627184428
tatc-dependent translocation of pyoverdine is responsible for the microbial growth suppression.infections are often not caused by a colonization of pseudomonas aeruginosa alone but by a consortium of other bacteria. little is known about the impact of p. aeruginosa on the growth of other bacteria upon coinfection. here, cell-ree culture supernatants obtained from p. aeruginosa suppressed the growth of a number of bacterial strains such as corynebacterium glutamicum, bacillus subtilis, staphylococcus aureus, and agrobacterium tumefaciens, but had little effect on the growth of escherichia ...201626832668
metabolic engineering of corynebacterium glutamicum for enhanced production of 5-aminovaleric acid.5-aminovaleric acid (5ava) is an important five-carbon platform chemical that can be used for the synthesis of polymers and other chemicals of industrial interest. enzymatic conversion of l-lysine to 5ava has been achieved by employing lysine 2-monooxygenase encoded by the davb gene and 5-aminovaleramidase encoded by the dava gene. additionally, a recombinant escherichia coli strain expressing the davb and dava genes has been developed for bioconversion of l-lysine to 5ava. to use glucose and xy ...201627717386
systems metabolic engineering of corynebacterium glutamicum for the production of the carbon-5 platform chemicals 5-aminovalerate and glutarate.the steadily growing world population and our ever luxurious life style, along with the simultaneously decreasing fossil resources has confronted modern society with the issue and need of finding renewable routes to accommodate for our demands. shifting the production pipeline from raw oil to biomass requires efficient processes for numerous platform chemicals being produced with high yield, high titer and high productivity.201627618862
transcriptome and gene ontology (go) enrichment analysis reveals genes involved in biotin metabolism that affect l-lysine production in corynebacterium glutamicum.corynebacterium glutamicum is widely used for amino acid production. in the present study, 543 genes showed a significant change in their mrna expression levels in l-lysine-producing c. glutamicum atcc21300 than that in the wild-type c. glutamicum atcc13032. among these 543 differentially expressed genes (degs), 28 genes were up- or downregulated. in addition, 454 degs were functionally enriched and categorized based on blast sequence homologies and gene ontology (go) annotations using the blast ...201627005618
fast mechanically driven daughter cell separation is widespread in actinobacteria.dividing cells of the coccoid gram-positive bacterium staphylococcus aureus undergo extremely rapid (millisecond) daughter cell separation (dcs) driven by mechanical crack propagation, a strategy that is very distinct from the gradual, enzymatically driven cell wall remodeling process that has been well described in several rod-shaped model bacteria. to determine if other bacteria, especially those in the same phylum (firmicutes) or with similar coccoid shapes as s. aureus, might use a similar m ...201627578753
engineering corynebacterium glutamicum for violacein hyper production.corynebacterium glutamicum was used as a metabolic engineering chassis for production of crude violacein (mixture of violacein and deoxyviolacein) due to corynebacterium's gras status and advantages in tryptophan fermentation. the violacein is a commercially potential compound with various applications derived from l-tryptophan.201627557730
regulation of coenzyme a biosynthesis in the hyperthermophilic bacterium thermotoga maritima.regulation of coenzyme a (coa) biosynthesis in bacteria and eukaryotes occurs through feedback inhibition targeting type i and type ii pantothenate kinase (pank), respectively. in contrast, the activity of type iii pank is not affected by coa. as the hyperthermophilic bacterium thermotoga maritima harbors only a single type iii pank (tm-pank), here we examined the mechanisms that regulate coa biosynthesis in this organism. we first examined the enzyme responsible for the ketopantoate reductase ( ...201627161115
rapid electron transfer within the iii-iv supercomplex in corynebacterium glutamicum.complex iii in c. glutamicum has an unusual di-heme cyt. c1 and it co-purifies with complex iv in a supercomplex. here, we investigated the kinetics of electron transfer within this supercomplex and in the cyt. aa3 alone (cyt. bc1 was removed genetically). in the reaction of the reduced cyt. aa3 with o2, we identified the same sequence of events as with other a-type oxidases. however, even though this reaction is associated with proton uptake, no ph dependence was observed in the kinetics. for t ...201627682138
structural insights into a novel class of aspartate aminotransferase from corynebacterium glutamicum.aspartate aminotransferase from corynebacterium glutamicum (cgaspat) is a plp-dependent enzyme that catalyzes the production of l-aspartate and α-ketoglutarate from l-glutamate and oxaloacetate in l-lysine biosynthesis. in order to understand the molecular mechanism of cgaspat and compare it with those of other aspartate aminotransferases (aspats) from the aminotransferase class i, we determined the crystal structure of cgaspat. cgaspat functions as a dimer, and the cgaspat monomer consists of t ...201627355211
regulons of global transcription factors in corynebacterium glutamicum.corynebacterium glutamicum, a high gc content gram-positive soil bacterium in actinobacteria, has been used for the industrial production of amino acids and engineered to produce various compounds, including polymer building blocks and biofuels. since its genome sequence was first published, its versatile metabolic pathways and their genetic components and regulatory mechanisms have been extensively studied. previous studies on transcriptional factors, including two-component systems and σ facto ...201626496920
biosensor-driven adaptive laboratory evolution of l-valine production in corynebacterium glutamicum.adaptive laboratory evolution has proven a valuable strategy for metabolic engineering. here, we established an experimental evolution approach for improving microbial metabolite production by imposing an artificial selective pressure on the fluorescent output of a biosensor using fluorescence-activated cell sorting. cells showing the highest fluorescent output were iteratively isolated and (re-)cultivated. the l-valine producer corynebacterium glutamicum δacee was equipped with an l-valine-resp ...201526453945
metabolic engineering of corynebacterium glutamicum strain atcc13032 to produce l-methionine.l-methionine-producing strain qw102/pjyw-4-hom(m) -lysc(m) -brnfe was developed from corynebacterium glutamicum strain atcc13032, using metabolic engineering strategies. these strategies involved (i) deletion of the gene thrb encoding homoserine kinase to increase the precursor supply, (ii) deletion of the gene mcbr encoding the regulator mcbr to release the transcriptional repression to various genes in the l-methionine biosynthetic pathway, (iii) overexpression of the gene lysc(m) encoding fee ...201525196586
effect of tween 40 and dtsr1 on l-arginine overproduction in corynebacterium crenatum.l-glutamate is an important precursor in the l-arginine (l-arg) biosynthetic pathway. various methods, including polyoxyethylene sorbitan monopalmitate (tween 40) addition and dtsr1 disruption, have been widely used to induce l-glutamate overproduction in corynebacterium glutamicum. in this study, a novel strategy for l-arg overproduction through tween 40 trigger and δdtsr1 mutant were proposed in corynebacterium crenatum.201526264811
regulation of the expression of de novo pyrimidine biosynthesis genes in corynebacterium glutamicum.expression of pyrimidine de novo biosynthesis is downregulated by an exogenous uracil in many bacteria. in this study, we show that a putative binding motif sequence of pyrr is required for uracil-mediated repression of pyrr-lacz translational fusion. however, the uracil response was still observed in the strain with the pyrr gene deleted, implying the existence of a uracil response factor other than pyrr which also acts through the pyrr binding loop region. deletion of rho, encoding the transcr ...201526260458
exploring the role of sigma factor gene expression on production by corynebacterium glutamicum: sigma factor h and fmn as example.bacteria are known to cope with environmental changes by using alternative sigma factors binding to rna polymerase core enzyme. sigma factor is one of the targets to modify transcription regulation in bacteria and to influence production capacities. in this study, the effect of overexpressing each annotated sigma factor gene in corynebacterium glutamicum wt was assayed using an iptg inducible plasmid system and different iptg concentrations. it was revealed that growth was severely decreased whe ...201526257719
trna-dependent alanylation of diacylglycerol and phosphatidylglycerol in corynebacterium glutamicum.aminoacyl-phosphatidylglycerol synthases (aapgss) are membrane proteins that utilize aminoacylated trnas to modify membrane lipids with amino acids. aminoacylation of membrane lipids alters the biochemical properties of the cytoplasmic membrane and enables bacteria to adapt to changes in environmental conditions. aapgss utilize alanine, lysine and arginine as modifying amino acids, and the primary lipid recipients have heretofore been defined as phosphatidylglycerol (pg) and cardiolipin. here we ...201526235234
live cell imaging of sos and prophage dynamics in isogenic bacterial populations.almost all bacterial genomes contain dna of viral origin, including functional prophages or degenerated phage elements. a frequent but often unnoted phenomenon is the spontaneous induction of prophage elements (spi) even in the absence of an external stimulus. in this study, we have analyzed spi of the large, degenerated prophage cgp3 (187 kbp), which is integrated into the genome of the gram-positive corynebacterium glutamicum atcc 13032. time-lapse fluorescence microscopy of fluorescent report ...201526235130
engineering microbial cell factories: metabolic engineering of corynebacterium glutamicum with a focus on non-natural products.corynebacterium glutamicum is the workhorse of biotechnological amino acid production. for more than 50 years amino acid producing strains of this actinomycete have been improved by classical breeding, metabolic engineering and systems and synthetic biology approaches. this review focusses mainly on recent developments on c. glutamicum strain development for non-natural products. recently, metabolite sensors have accelerated classical strain breeding. synthetic pathways for access to alternative ...201526216246
co-production of s-adenosyl-l-methionine and l-isoleucine in corynebacterium glutamicum.in this study, production of s-adenosyl-l-methionine in corynebacterium glutamicum was investigated by overexpressing genes metk and vgb. compared with vector control, overexpression of metk alone in c. glutamicum atcc13032 and iwj001 increased sam production 5.11 and 11.65 times, respectively; while overexpression of metk and vgb in c. glutamicum atcc13032 and iwj001 increased sam production 5.83 and 14.95 times, respectively. further studies on iwj001/pdxw-8-metk-vgb showed that the limiting f ...201526215341
metabolic engineering of corynebacterium glutamicum for the production of l-ornithine.l-ornithine is a non-essential amino acid for various industrial applications in food industry. in this study, corynebacterium glutamicum atcc 13032 was metabolically engineered for the production of l-ornithine. first, the prob and argf genes were deleted to block the competitive branch pathway and to block the conversion of l-ornithine to citrulline, respectively. in addition, the argr gene encoding the regulatory repressor of the l-arginine operon was also deleted. the resulting strain produc ...201525163446
rational design of allosteric regulation of homoserine dehydrogenase by a nonnatural inhibitor l-lysine.allosteric proteins, which can sense different signals, are interesting biological parts for synthetic biology. in particular, the design of an artificial allosteric enzyme to sense an unnatural signal is both challenging and highly desired, for example, for a precise and dynamical control of fluxes of growth-essential but byproduct pathways in metabolic engineering of industrial microorganisms. in this work, we used homoserine dehydrogenase (hsdh) of corynebacterium glutamicum, which is natural ...201524344690
exploring signal transduction in heteromultimeric protein based on energy dissipation model.dynamic intersubunit interactions are key elements in the regulation of many biological systems. a better understanding of how subunits interact with each other and how their interactions are related to dynamic protein structure is a fundamental task in biology. in this paper, a heteromultimeric allosteric protein, corynebacterium glutamicum aspartokinase, is used as a model system to explore the signal transduction involved in intersubunit interactions and allosteric communication with an empha ...201524279729
chassis organism from corynebacterium glutamicum--a top-down approach to identify and delete irrelevant gene clusters.for synthetic biology applications, a robust structural basis is required, which can be constructed either from scratch or in a top-down approach starting from any existing organism. in this study, we initiated the top-down construction of a chassis organism from corynebacterium glutamicum atcc 13032, aiming for the relevant gene set to maintain its fast growth on defined medium. we evaluated each native gene for its essentiality considering expression levels, phylogenetic conservation, and knoc ...201525139579
metabolic engineering of corynebacterium glutamicum for methanol metabolism.methanol is already an important carbon feedstock in the chemical industry, but it has found only limited application in biotechnological production processes. this can be mostly attributed to the inability of most microbial platform organisms to utilize methanol as a carbon and energy source. with the aim to turn methanol into a suitable feedstock for microbial production processes, we engineered the industrially important but nonmethylotrophic bacterium corynebacterium glutamicum toward the ut ...201525595770
functional characterization of a vanillin dehydrogenase in corynebacterium glutamicum.vanillin dehydrogenase (vdh) is a crucial enzyme involved in the degradation of lignin-derived aromatic compounds. herein, the vdh from corynebacterium glutamicum was characterized. the relative molecular mass (mr) determined by sds-page was ~51 kda, whereas the apparent native mr values revealed by gel filtration chromatography were 49.5, 92.3, 159.0 and 199.2 kda, indicating the presence of dimeric, trimeric and tetrameric forms. moreover, the enzyme showed its highest level of activity toward ...201525622822
biosynthesis of l-sorbose and l-psicose based on c-c bond formation catalyzed by aldolases in an engineered corynebacterium glutamicum strain.the property of loose stereochemical control at aldol products from aldolases helped to synthesize multiple polyhydroxylated compounds with nonnatural stereoconfiguration. in this study, we discovered for the first time that some fructose 1,6-diphosphate aldolases (frua) and tagatose 1,6-diphosphate (taga) aldolases lost their strict stereoselectivity when using l-glyceraldehyde and synthesized not only l-sorbose but also a high proportion of l-psicose. among the aldolases tested, taga from baci ...201525888171
ohr protects corynebacterium glutamicum against organic hydroperoxide induced oxidative stress.ohr, a bacterial protein encoded by the organic hydroperoxide resistance (ohr) gene, plays a critical role in resistance to organic hydroperoxides. in the present study, we show that the cys-based thiol-dependent ohr of corynebacterium glutamicum decomposes organic hydroperoxides more efficiently than hydrogen peroxide. replacement of either of the two cys residues of ohr by a ser residue resulted in drastic loss of activity. the electron donors supporting regeneration of the peroxidase activity ...201526121694
interaction sites of diviva and roda from corynebacterium glutamicum.elongation growth in actinobacteria is localized at the cell poles. this is in contrast to many classical model organisms where insertion of new cell wall material is localized around the lateral site. we previously described a role of roda from corynebacterium glutamicum in apical cell growth and morphogenesis. deletion of roda had drastic effects on morphology and growth, likely a result from misregulation of penicillin-binding proteins and cell wall precursor delivery. we identified the inter ...201525709601
redox regulation by reversible protein s-thiolation in bacteria.low molecular weight (lmw) thiols function as thiol-redox buffers to maintain the reduced state of the cytoplasm. the best studied lmw thiol is the tripeptide glutathione (gsh) present in all eukaryotes and gram-negative bacteria. firmicutes bacteria, including bacillus and staphylococcus species utilize the redox buffer bacillithiol (bsh) while actinomycetes produce the related redox buffer mycothiol (msh). in eukaryotes, proteins are post-translationally modified to s-glutathionylated proteins ...201525852656
engineering the glycolytic pathway: a potential approach for improvement of biocatalyst performance.the glycolytic pathway is a main driving force in the fermentation process as it produces energy, cell component precursors, and fermentation products. given its importance, the glycolytic pathway can be considered as an attractive target for the metabolic engineering of industrial microorganisms. however, many attempts to enhance glycolytic flux, by overexpressing homologous or heterologous genes encoding glycolytic enzymes, have been unsuccessful. in contrast, significant enhancement in glycol ...201526513591
metabolic engineering of an atp-neutral embden-meyerhof-parnas pathway in corynebacterium glutamicum: growth restoration by an adaptive point mutation in nadh dehydrogenase.corynebacterium glutamicum uses the embden-meyerhof-parnas pathway of glycolysis and gains 2 mol of atp per mol of glucose by substrate-level phosphorylation (slp). to engineer glycolysis without net atp formation by slp, endogenous phosphorylating nad-dependent glyceraldehyde-3-phosphate dehydrogenase (gapdh) was replaced by nonphosphorylating nadp-dependent glyceraldehyde-3-phosphate dehydrogenase (gapn) from clostridium acetobutylicum, which irreversibly converts glyceraldehyde-3-phosphate (g ...201525576602
identification of two mutations increasing the methanol tolerance of corynebacterium glutamicum.methanol is present in most ecosystems and may also occur in industrial applications, e.g. as an impurity of carbon sources such as technical glycerol. methanol often inhibits growth of bacteria, thus, methanol tolerance may limit fermentative production processes.201526474849
reactions of cg10062, a cis-3-chloroacrylic acid dehalogenase homologue, with acetylene and allene substrates: evidence for a hydration-dependent decarboxylation.cg10062 is a cis-3-chloroacrylic acid dehalogenase (cis-caad) homologue from corynebacterium glutamicum with an unknown function and an uninformative genomic context. it shares 53% pairwise sequence similarity with cis-caad including the six active site amino acids (pro-1, his-28, arg-70, arg-73, tyr-103, and glu-114) that are critical for cis-caad activity. however, cg10062 is a poor cis-caad: it lacks catalytic efficiency and isomer specificity. two acetylene compounds (propiolate and 2-butyno ...201525894805
unraveling the kinetic diversity of microbial 3-dehydroquinate dehydratases of shikimate pathway.3-dehydroquinate dehydratase (dhqase) catalyzes the conversion of 3-dehydroquinic acid to 3-dehydroshikimic acid of the shikimate pathway. in this study, 3180 prokaryotic genomes were examined and 459 dhqase sequences were retrieved. based on sequence analysis and their original hosts, 38 dhqase genes were selected for chemical synthesis. the selected dhqases were translated into new dna sequences according to the genetic codon usage bias by both escherichia coli and corynebacterium glutamicum. ...201525852984
screening currency notes for microbial pathogens and antibiotic resistance genes using a shotgun metagenomic approach.fomites are a well-known source of microbial infections and previous studies have provided insights into the sojourning microbiome of fomites from various sources. paper currency notes are one of the most commonly exchanged objects and its potential to transmit pathogenic organisms has been well recognized. approaches to identify the microbiome associated with paper currency notes have been largely limited to culture dependent approaches. subsequent studies portrayed the use of 16s ribosomal rna ...201526035208
single-domain peptidyl-prolyl cis/trans isomerase fkpa from corynebacterium glutamicum improves the biomass yield at increased growth temperatures.peptidyl-prolyl cis/trans isomerases (ppiases) catalyze the rate-limiting protein folding step at peptidyl bonds preceding proline residues and were found to be involved in several biological processes, including gene expression, signal transduction, and protein secretion. representative enzymes were found in almost all sequenced genomes, including corynebacterium glutamicum, a facultative anaerobic gram-positive and industrial workhorse for the production of amino acids. in c. glutamicum, a pre ...201526341203
metabolic engineering of corynebacterium glutamicum for the production of itaconate.the capability of corynebacterium glutamicum for glucose-based synthesis of itaconate was explored, which can serve as building block for production of polymers, chemicals, and fuels. c. glutamicum was highly tolerant to itaconate and did not metabolize it. expression of the aspergillus terreus cad1 gene encoding cis-aconitate decarboxylase (cad) in strain atcc13032 led to the production of 1.4mm itaconate in the stationary growth phase. fusion of cad with the escherichia coli maltose-binding pr ...201526100077
metabolic engineering of itaconate production in escherichia coli.interest in sustainable development has led to efforts to replace petrochemical-based monomers with biomass-based ones. itaconic acid, a c5-dicarboxylic acid, is a potential monomer for the chemical industry with many prospective applications. cis-aconitate decarboxylase (cada) is the key enzyme of itaconate production, converting the citric acid cycle intermediate cis-aconitate into itaconate. heterologous expression of cada from aspergillus terreus in escherichia coli resulted in low cada acti ...201525277412
high-level production of bacillus cereus phospholipase c in corynebacterium glutamicum.enzymatic oil degumming (removal of phospholipids) using phospholipase c (plc) is a well-established and environmentally friendly process for vegetable oil refining. in this work, we report the production of recombinant bacillus cereus plc in corynebacterium glutamicum atcc 13869 in a high cell density fermentation process and its performance in soybean oil degumming. a final concentration of 5.5g/l of the recombinant enzyme was achieved when the respective gene was expressed from the tac promot ...201526519562
a prophage-encoded actin-like protein required for efficient viral dna replication in bacteria.in host cells, viral replication is localized at specific subcellular sites. viruses that infect eukaryotic and prokaryotic cells often use host-derived cytoskeletal structures, such as the actin skeleton, for intracellular positioning. here, we describe that a prophage, cgp3, integrated into the genome of corynebacterium glutamicum encodes an actin-like protein, alpc. biochemical characterization confirms that alpc is a bona fide actin-like protein and cell biological analysis shows that alpc f ...201525916847
corynebacterium glutamicum methionine sulfoxide reductase a uses both mycoredoxin and thioredoxin for regeneration and oxidative stress resistance.oxidation of methionine leads to the formation of the s and r diastereomers of methionine sulfoxide (meto), which can be reversed by the actions of two structurally unrelated classes of methionine sulfoxide reductase (msr), msra and msrb, respectively. although msras have long been demonstrated in numerous bacteria, their physiological and biochemical functions remain largely unknown in actinomycetes. here, we report that a corynebacterium glutamicum methionine sulfoxide reductase a (cgmsra) tha ...201525681179
engineering corynebacterium glutamicum for the production of 2,3-butanediol.2,3-butanediol is an important bulk chemical with a wide range of applications. in bacteria, this metabolite is synthesised from pyruvate via a three-step pathway involving α-acetolactate synthase, α-acetolactate decarboxylase and 2,3-butanediol dehydrogenase. thus far, the best producers of 2,3-butanediol are pathogenic strains, hence, the development of more suitable organisms for industrial scale fermentation is needed. herein, 2,3-butanediol production was engineered in the generally regarde ...201526511723
metabolic pathway engineering for production of 1,2-propanediol and 1-propanol by corynebacterium glutamicum.production of the versatile bulk chemical 1,2-propanediol and the potential biofuel 1-propanol is still dependent on petroleum, but some approaches to establish bio-based production from renewable feed stocks and to avoid toxic intermediates have been described. the biotechnological workhorse corynebacterium glutamicum has also been shown to be able to overproduce 1,2-propanediol by metabolic engineering. additionally, c. glutamicum has previously been engineered for production of the biofuels e ...201526110019
[characterization of l-aspartate-α-decarboxylase from bacillus subtilis].as an important material in pharmaceutical and chemical industry, β-alanine was mainly produced by chemical methods. l-aspartate-α-decarboxylase could catalyze the α-decarboxylation from l-aspartate to β-alanine. determinations for specific activities of pands from escherichia coli, corynebacterium glutamicum and bacillus subtilis were performed in this study (0.98 u/mg, 7.52 u/mg and 8.4 u/mg respectively). the optimal temperature and ph of pands from c. glutamicum and b. subtilis were 65 degre ...201526762040
a tatabc-type tat translocase is required for unimpaired aerobic growth of corynebacterium glutamicum atcc13032.the twin-arginine translocation (tat) system transports folded proteins across the cytoplasmic membrane of bacteria and the thylakoid membrane of plant chloroplasts. escherichia coli and other gram-negative bacteria possess a tatabc-type tat translocase in which each of the three inner membrane proteins tata, tatb, and tatc performs a mechanistically distinct function. in contrast, low-gc gram-positive bacteria, such as bacillus subtilis, use a tatac-type minimal tat translocase in which the tat ...201525837592
exploring lysine riboswitch for metabolic flux control and improvement of l-lysine synthesis in corynebacterium glutamicum.riboswitch, a regulatory part of an mrna molecule that can specifically bind a metabolite and regulate gene expression, is attractive for engineering biological systems, especially for the control of metabolic fluxes in industrial microorganisms. here, we demonstrate the use of lysine riboswitch and intracellular l-lysine as a signal to control the competing but essential metabolic by-pathways of lysine biosynthesis. to this end, we first examined the natural lysine riboswitches of eschericia co ...201525575181
metabolic engineering of the mixed-acid fermentation pathway of escherichia coli for anaerobic production of glutamate and itaconate.itaconic acid, an unsaturated c5-dicarboxylic acid, is a biobased building block for the polymer industry. the purpose of this study was to establish proof of principle for an anaerobic fermentation process for the production of itaconic acid by modification of the mixed acid fermentation pathway of e. coli. e. coli bw25113 (de3) and the phosphate acetyltransferase (pta) and lactate dehydrogenase (ldha) deficient strain e. coli bw25113 (de3) δpta-δldha were used to study anaerobic itaconate prod ...201526384341
engineering of corynebacterium glutamicum for growth and succinate production from levoglucosan, a pyrolytic sugar substrate.thermochemical processing provides continuous production of bio-oils from lignocellulosic biomass. levoglucosan, a pyrolytic sugar substrate c6h10o5 in a bio-oil, has been used for ethanol production using engineered escherichia coli. here we provide the first example for succinate production from levoglucosan with corynebacterium glutamicum, a well-known industrial amino acid producer. heterologous expression of a gene encoding a sugar kinase from lipomyces starkeyi, gibberella zeae or pseudomo ...201526363018
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