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Sample records for black-pigmented corynebacterium aurimucosum

  1. Complete genome sequence and lifestyle of black-pigmented Corynebacterium aurimucosum ATCC 700975 (formerly C. nigricans CN-1 isolated from a vaginal swab of a woman with spontaneous abortion

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    Gartemann Karl-Heinz

    2010-02-01

    Full Text Available Abstract Background Corynebacterium aurimucosum is a slightly yellowish, non-lipophilic, facultative anaerobic member of the genus Corynebacterium and predominantly isolated from human clinical specimens. Unusual black-pigmented variants of C. aurimucosum (originally named as C. nigricans continue to be recovered from the female urogenital tract and they are associated with complications during pregnancy. C. aurimucosum ATCC 700975 (C. nigricans CN-1 was originally isolated from a vaginal swab of a 34-year-old woman who experienced a spontaneous abortion during month six of pregnancy. For a better understanding of the physiology and lifestyle of this potential urogenital pathogen, the complete genome sequence of C. aurimucosum ATCC 700975 was determined. Results Sequencing and assembly of the C. aurimucosum ATCC 700975 genome yielded a circular chromosome of 2,790,189 bp in size and the 29,037-bp plasmid pET44827. Specific gene sets associated with the central metabolism of C. aurimucosum apparently provide enhanced metabolic flexibility and adaptability in aerobic, anaerobic and low-pH environments, including gene clusters for the uptake and degradation of aromatic amines, L-histidine and L-tartrate as well as a gene region for the formation of selenocysteine and its incorporation into formate dehydrogenase. Plasmid pET44827 codes for a non-ribosomal peptide synthetase that plays the pivotal role in the synthesis of the characteristic black pigment of C. aurimucosum ATCC 700975. Conclusions The data obtained by the genome project suggest that C. aurimucosum could be both a resident of the human gut and possibly a pathogen in the female genital tract causing complications during pregnancy. Since hitherto all black-pigmented C. aurimucosum strains have been recovered from female genital source, biosynthesis of the pigment is apparently required for colonization by protecting the bacterial cells against the high hydrogen peroxide concentration in

  2. Black-pigmented sputum

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    Rafael Martínez-Girón

    2013-01-01

    Full Text Available Black-pigmented sputum, also called "melanoptysis," is a symptom that may be observed in certain pathologies such us coal workers′ pneumoconiosis (anthracosis. The cavitation and liquefaction of a fibrosed mass by an infectious process (tuberculosis, infections by anaerobes, etc. or by ischemic necrosis may cause expectoration of a blackish secretion. We report the case of a patient with labor precedents as a coal worker, from whom abundant black sputum was obtained in the course of an incidental expectoration. Cyto-histological findings are shown and a differential diagnosis is established.

  3. In vitro activity of tigecycline and 10 other antimicrobials against clinical isolates of the genus Corynebacterium.

    Science.gov (United States)

    Fernandez-Roblas, R; Adames, H; Martín-de-Hijas, N Z; Almeida, D García; Gadea, I; Esteban, J

    2009-05-01

    We studied the in vitro activity of tigecycline and 10 other commonly used antibiotics against 135 clinical isolates of non-diphtheria Corynebacterium spp. using the Etest system. Tigecycline minimum inhibitory concentrations for 50% and 90% of the organisms (MIC(50) and MIC(90) values, respectively, in mg/L) were: Corynebacterium urealyticum, 0.094 and 0.125; Corynebacterium amycolatum, 0.125 and 2; Corynebacterium jeikeium, 0.094 and 0.75; Corynebacterium coyleae, 0.064 and 0.064; Corynebacterium striatum, 0.064 and 1; Corynebacterium aurimucosum, 0.094 and 0.125; and Corynebacterium afermentans, 0.064 and 0.094. The activities of all other antimicrobials were variable, with good activity of glycopeptides, linezolid, quinupristin/dalfopristin and daptomycin and with resistance to macrolides in a high number of isolates. Tigecycline is a good alternative for the therapy of infections caused by non-diphtheria corynebacteria. PMID:19153032

  4. Characterization of ancient Chinese pottery decorated with a black pigment

    International Nuclear Information System (INIS)

    The Yangshao type pottery, made about 6000 yrs ago, was investigated by X-ray diffraction (XRD), and confirmed to be composed of quartz, feldspar, muscovite and calcite. A black pigment on it was assumed to be (Mn, Fe)3O4 from Particle Induced X-ray Emission (PIXE) and XRD experiments. Firing temperature of the pottery was assumed to be less than 600 deg. C from a heating experiment of the fragment of the pottery

  5. Long-wave UV light fluorescence for identification of black-pigmented Bacteroides spp.

    OpenAIRE

    Slots, J; Reynolds, H S

    1982-01-01

    Black-pigmented Bacteroides strains were grown on blood agar, and the colonies were evaluated for fluorescence from long-wave UV light. Most test strains of Bacteroides melaninogenicus subsp. intermedius exhibited a brilliant red fluorescence. B. melaninogenicus subsp. melaninogenicus fluoresced mostly red-orange. Bacteroides asaccharolyticus showed a yellow or red fluorescence. The intensity of the Bacteroides fluorescence weakened when the black pigment of the colonies developed. In contras...

  6. Synthesis and Application of Carbon–Iron Oxide Microspheres’ Black Pigments in Electrophoretic Displays

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    Meng Xianwei

    2010-01-01

    Full Text Available Abstract Carbon–iron oxide microspheres’ black pigments (CIOMBs had been prepared via ultrasonic spray pyrolysis of aqueous solutions containing ferrous chloride and glucose. Due to the presence of carbon, CIOMBs not only exhibited remarkably acid resistance, but also could be well dispersed in both polar solvents and nonpolar solvent. Finally, dispersions of hollow CIOMBs in tetrachloroethylene had successfully been applied in electrophoretic displays.

  7. Black pigments of rock art: identification of inorganic and organic components by combining analytical techniques

    International Nuclear Information System (INIS)

    Archaeological samples are complex in composition since they generally comprise a mixture of materials submitted to deterioration factors largely dependent on the environmental conditions. Therefore, the integration of analytical tools such as TXRF, FT-IR and GC-MS can maximize the amount of information provided by the sample. Recently, two black rock art samples of camelid figures at Alero Hornillos 2, an archaeological site located near the town of Susques (Jujuy Province, Argentina), were investigated. TXRF technique, selected for inorganic information, as well as FT-IR and GC-MS were employed in order to discover inorganic and organic composition of the black pigments. (author)

  8. New research on the preparation of the black pigment from vaccinium bracteatum thunb%关于乌饭树黑色素制备的新思路

    Institute of Scientific and Technical Information of China (English)

    魏国华; 邵斌; 许新德

    2011-01-01

    乌饭树黑色素的研究虽然已经进行了多年,但其制备过程却一直没有受到应有的重视,从而使其成为制约乌饭树黑色素产业化的一个瓶颈.本文从乌饭树黑色素制备过程中的浸提、氧化发酵、纯化和干燥四个方面进行了必要的分析,并提出了一些较为新颖的观点:第一,氧化发酵过程是乌饭树黑色素形成的必要条件;第二,树脂是纯化乌饭树黑色素的更好选择;第三,不能用黄酮类物质的含量来度量乌饭树黑色素的含量.希望这些观点能为乌饭树黑色素的产业化提供一点新思路.%The study on the black pigment from Vaccinium Bracteatum Thunb has been investigated for many years. However, the preparation has not been given sufficient attention, thus prevents its industrialization. The process of preparation has been analyzed in this paper. It showed that; the oxidation was the necessary condition for preparing black pigment from Vaccinium Bracteatum Thunb; moreover, the resin is a good choice to purify the black pigment from Vaccinium Bracteatum Thunb; and the last, it was not appropriate to use the content of total flavonoids to evaluate the content of black pigment from Vaccinium Bracteatum Thunb.

  9. Skin Infections Due to Corynebacterium

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    Meltem Türkmen

    2010-03-01

    Full Text Available Corynebacteria are Gram-positive, non-sporulated, non-capsulated, aerobic diphtheroid bacteria accounting for nearly 50%of the natural skin biocene. This bacterial family is responsible for various skin diseases such as cutaneous diphteria, cromhydrosis, bromhydrosis but the most common of them are pitted keratolysis, trichobacteriosis and erythrasma. A warm and moist environment and poor hygiene are the predisposition factors for these three diseases. Although this skin diseases are seen more frequently, they usually mistaken for a mycotic infection by general practitioners, with subsequent antimycotic treatment. Here skin diseases compromised with Corynebacterium are presented with their demographic features and discussed on the basis of a literature review.

  10. Intrapleural Corynebacterium parvum for malignant pleural effusions

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    R. Felletti; Ravazzoni, C

    1983-01-01

    A pilot study of topical (intrapleural) treatment with Corynebacterium parvum was carried out in 10 patients with malignant pleural effusions complicating primary or secondary neoplasms and necessitating frequent thoracocentesis for symptomatic relief. The method was aspiration of all intrapleural fluid except a small portion left for dilution, and then injection of 7 mg of a preparation of Corynebacterium parvum suspended in 20 ml of normal saline solution. The treatment was repeated in each...

  11. Production of diphtheria toxin by selected isolates of Corynebacterium ulcerans and Corynebacterium pseudotuberculosis.

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    Wong, T P; Groman, N

    1984-01-01

    We determined the diphtheria toxin phenotype of specially selected isolates of Corynebacterium ulcerans and Corynebacterium pseudotuberculosis (C. ovis). All produced proteins similar in size and immunological structure to diphtheria toxin. As with diphtheria toxin, they exhibited ADP-ribosylating activity, and their synthesis was regulated by iron.

  12. Spontaneous corynebacterium spondylodiskitis in an immunocompetent patient

    DEFF Research Database (Denmark)

    Sharifi, Guive; Hosseinzadeh Bakhtevari, Mehrdad; Nabizadeh, Naveed;

    2016-01-01

    Background Spontaneous Corynebacterium spondylodiskitis is an unusual diagnosis of spondylodiskitis, especially in healthy patients without any significant past medical history. Materials and methods We describe the case of a 78-year-old man with progressive low back pain for 3 months, irradiating...... down the lower limbs through L5 and S1 root pathways, associated with distal muscle weakness in both lower limbs. He had no history of trauma or medical problems. Laboratory investigation revealed elevated serum C-reactive protein (CRP) and erythrocyte sedimentation rate, without leukocytosis...... Corynebacterium infection. Results One month after surgery, the pain diminished dramatically and the CRP titer diminished significantly. Conclusion Although cases are very rare, spontaneous Corynebacterium spondylodiskitis, with substantial invasion of the spine, may develop in patients lacking any history...

  13. Corynebacterium haemolyticum infections in Sri Lanka.

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    Wickremesinghe, R. S.

    1981-01-01

    Corynebacterium haemolyticum infections are described for the first time in Sri Lanka. In a period of 2 years from 1978-80 C. haemolyticum was isolated from the pharynx of 9 patients wih tonsillitis and from local septic lesions in 7 other patients. Association with other pathogens was common. No patients had a rash. The properties of the isolates are described.

  14. Corynebacterium macginleyi isolated from a corneal ulcer

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    Kathryn Ruoff

    2010-02-01

    Full Text Available We report the isolation of Corynebacterium macginleyi from the corneal ulcer culture of a patient, later enrolled in the Steroids for Corneal Ulcer Trial (SCUT. To our knowledge this is the first published report from North America of the recovery of C. macginleyi from a serious ocular infection.

  15. Corynebacterium ulcerans, an emerging human pathogen.

    Science.gov (United States)

    Hacker, Elena; Antunes, Camila A; Mattos-Guaraldi, Ana L; Burkovski, Andreas; Tauch, Andreas

    2016-09-01

    While formerly known infections of Corynebacterium ulcerans are rare and mainly associated with contact to infected cattle, C. ulcerans has become an emerging pathogen today. In Western Europe, cases of respiratory diphtheria caused by C. ulcerans have been reported more often than infections by Corynebacterium diphtheria, while systemic infections are also increasingly reported. Little is known about factors that contribute to host colonization and virulence of this zoonotic pathogen. Research in this field has received new impetus by the publication of several C. ulcerans genome sequences in the past years. This review gives a comprehensive overview of the basic knowledge of C. ulcerans, as well as the recent advances made in the analysis of putative virulence factors.

  16. Corynebacterium ulcerans, an emerging human pathogen.

    Science.gov (United States)

    Hacker, Elena; Antunes, Camila A; Mattos-Guaraldi, Ana L; Burkovski, Andreas; Tauch, Andreas

    2016-09-01

    While formerly known infections of Corynebacterium ulcerans are rare and mainly associated with contact to infected cattle, C. ulcerans has become an emerging pathogen today. In Western Europe, cases of respiratory diphtheria caused by C. ulcerans have been reported more often than infections by Corynebacterium diphtheria, while systemic infections are also increasingly reported. Little is known about factors that contribute to host colonization and virulence of this zoonotic pathogen. Research in this field has received new impetus by the publication of several C. ulcerans genome sequences in the past years. This review gives a comprehensive overview of the basic knowledge of C. ulcerans, as well as the recent advances made in the analysis of putative virulence factors. PMID:27545005

  17. Corynebacterium glutamicum promoters: a practical approach

    OpenAIRE

    Pátek, M. (Miroslav); Holátko, J. (Jiří); Busche, T.; Kalinowski, J; Nešvera, J. (Jan)

    2013-01-01

    Summary 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...

  18. Synthetic promoter libraries for Corynebacterium glutamicum

    DEFF Research Database (Denmark)

    Rytter, Jakob Vang; Helmark, Søren; Chen, Jun;

    2014-01-01

    The ability to modulate gene expression is an important genetic tool in systems biology and biotechnology. Here, we demonstrate that a previously published easy and fast PCR-based method for modulating gene expression in lactic acid bacteria is also applicable to Corynebacterium glutamicum. We...... promoter library (SPL) technology is convenient for modulating gene expression in C. glutamicum and should have many future applications, within basic research as well as for optimizing industrial production organisms....

  19. Kontrolle des Aminozuckerstoffwechsels in Corynebacterium glutamicum

    OpenAIRE

    Uhde, Andreas

    2014-01-01

    Aminozucker, wie Glucosamin oder N-Acetylglucosamin, stellen für biotechno-logische Prozesse mit Corynebacterium glutamicum eine alternativer Kohlenstoffquelle dar. Als Bestandteil von Chitin sind sie weit verbreitet, können jedoch von C. glutamicum nur sehr schlecht oder, wie im Fall von N-Acetylglucosamin, gar nicht umgesetzt werden. Diese Arbeit beschreibt die Identifikation und Charakterisierung von Proteinen des Aminozuckerkatabolismus, die Wirkungsweise des an der Expression beteiligten...

  20. The killing of macrophages by Corynebacterium ulcerans.

    Science.gov (United States)

    Hacker, Elena; Ott, Lisa; Schulze-Luehrmann, Jan; Lührmann, Anja; Wiesmann, Veit; Wittenberg, Thomas; Burkovski, Andreas

    2016-01-01

    Corynebacterium ulcerans is an emerging pathogen transmitted by a zoonotic pathway with a very broad host spectrum to humans. Despite rising numbers of infections and potentially fatal outcomes, data on the molecular basis of pathogenicity are scarce. In this study, the interaction of 2 C. ulcerans isolates - one from an asymptomatic dog, one from a fatal case of human infection - with human macrophages was investigated. C. ulcerans strains were able to survive in macrophages for at least 20 hours. Uptake led to delay of phagolysosome maturation and detrimental effects on the macrophages as deduced from cytotoxicity measurements and FACS analyses. The data presented here indicate a high infectious potential of this emerging pathogen.

  1. NMR study of Corynebacterium melassecola metabolism; Etude du metabolisme de corynebacterium melassecola par RMN

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    Rollin, C.; Morgant, V.; Guyonvarch, A. [Centre ORSAN, 91 - Les Ulis (France); Guerquin Kern, J.L. [Institut Curie, 91 - Orsay (France)

    1994-12-31

    Corynebacterium melassecola is a microorganism producing glutamic acid, an aminate acid used as food additive. Knowledge of its metabolism is essential for improving the phyla. A study is carried out on intracellular extracts with NMR spectrometry in order to determine certain glucose catabolism pathways using a partial isotopic enrichment with (1-{sup 13}C) or (6-{sup 13}C) glucose. Results demonstrate the particular metabolism of Corynebacteria. 2 tabs., 3 refs.

  2. Interdigital foot infections: Corynebacterium minutissimum and agents of superficial mycoses

    OpenAIRE

    Fatma Mutlu Sariguzel; A. Nedret Koc; Gülhan Yagmur; Elife Berk

    2014-01-01

    Interdigital foot infections are mostly caused initially by dermatophytes, yeasts and less frequently by bacteria. Erythrasma caused by Corynebacterium minutissimum can be confused with superficial mycoses. The aim of the study was to determine the prevalence of the etiologic agents of superficial mycoses and the frequency of Corynebacterium minutissimum in interdigital foot infections. All the samples obtained from the 121 patients with interdigital foot infections were examined directly wit...

  3. Intrapleural immunotherapy with Corynebacterium parvum in recurrent malignant pleural effusions.

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    Millar, J W; Hunter, A M; Horne, N. W.

    1980-01-01

    Twenty-one patients with proven recurrent malignant pleural effusions were randomly allocated to treatment groups receiving either intrapleural Corynebacterium parvum in a dose of 7 mg or intrapleural mustine (20 mg). The designated intrapleural therapy was repeated on one occasion if further pleural aspiration was required. Corynebacterium parvum (nine patients) proved superior to mustine (12 patients) in suppressing the reaccumulation of pleural fluid, and was associated with only minimal s...

  4. Implication ofCorynebacterium species in food’s contamination

    Institute of Scientific and Technical Information of China (English)

    Sana Alibi; Asma Ferjani; Jalel Boukadida

    2016-01-01

    Corynebacteriumspp. are part of the human microbiota. Recently, species of this genus are increasingly implicated in different types of infections especially in immunocompromized and hospitalized patients. The significance of the presence of the genusCorynebacterium in foods is not clearly established. These bacteria may be involved in spoilage or ripening of cheese and meats. This review focused on different researches concerning the implication of Corynebacterium species in food’s contamination.

  5. The Actinobacterium Corynebacterium glutamicum, an Industrial Workhorse.

    Science.gov (United States)

    Lee, Joo-Young; Na, Yoon-Ah; Kim, Eungsoo; Lee, Heung-Shick; Kim, Pil

    2016-05-28

    Starting as a glutamate producer, Corynebacterium glutamicum has played a variety of roles in the industrial production of amino acids, one of the most important areas of white biotechnology. From shortly after its genome information became available, C. glutamicum has been applied in various production processes for value-added chemicals, fuels, and polymers, as a key organism in industrial biotechnology alongside the surprising progress in systems biology and metabolic engineering. In addition, recent studies have suggested another potential for C. glutamicum as a synthetic biology platform chassis that could move the new era of industrial microbial biotechnology beyond the classical field. Here, we review the recent progress and perspectives in relation to C. glutamicum, which demonstrate it as one of the most promising and valuable workhorses in the field of industrial biotechnology.

  6. Staphylococcus aureus shifts towards commensalism in response to Corynebacterium species

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    Matthew M Ramsey

    2016-08-01

    Full Text Available Staphylococcus aureus–human interactions result in a continuum of outcomes from commensalism to pathogenesis. S. aureus is a clinically important pathogen that asymptomatically colonizes ~25% of humans as a member of the nostril and skin microbiota, where it resides with other bacteria including commensal Corynebacterium species. Commensal Corynebacterium spp. are also positively correlated with S. aureus in chronic polymicrobial diabetic foot infections, distinct from acute monomicrobial S. aureus infections. Recent work by our lab and others indicates that microbe-microbe interactions between S. aureus and human skin/nasal commensals, including Corynebacterium species, affect S. aureus behavior and fitness. Thus, we hypothesized that S. aureus interactions with Corynebacterium spp. diminish S. aureus virulence. We tested this by assaying for changes in S. aureus gene expression during in vitro mono- versus coculture with Corynebacterium striatum, a common skin and nasal commensal. We observed a broad shift in S. aureus gene transcription during in vitro growth with C. striatum, including increased transcription of genes known to exhibit increased expression during human nasal colonization and decreased transcription of virulence genes. S. aureus uses several regulatory pathways to transition between commensal and pathogenic states. One of these, the quorum signal accessory gene regulator (agr system, was strongly inhibited in response to Corynebacterium spp. Phenotypically, S. aureus exposed to C. striatum exhibited increased adhesion to epithelial cells, reflecting a commensal state, and decreased hemolysin activity, reflecting an attenuation of virulence. Consistent with this, S. aureus displayed diminished fitness in experimental in vivo coinfection with C. striatum when compared to monoinfection. These data support a model in which S. aureus shifts from virulence towards a commensal state when exposed to commensal Corynebacterium species.

  7. Staphylococcus aureus Shifts toward Commensalism in Response to Corynebacterium Species.

    Science.gov (United States)

    Ramsey, Matthew M; Freire, Marcelo O; Gabrilska, Rebecca A; Rumbaugh, Kendra P; Lemon, Katherine P

    2016-01-01

    Staphylococcus aureus-human interactions result in a continuum of outcomes from commensalism to pathogenesis. S. aureus is a clinically important pathogen that asymptomatically colonizes ~25% of humans as a member of the nostril and skin microbiota, where it resides with other bacteria including commensal Corynebacterium species. Commensal Corynebacterium spp. are also positively correlated with S. aureus in chronic polymicrobial diabetic foot infections, distinct from acute monomicrobial S. aureus infections. Recent work by our lab and others indicates that microbe-microbe interactions between S. aureus and human skin/nasal commensals, including Corynebacterium species, affect S. aureus behavior and fitness. Thus, we hypothesized that S. aureus interactions with Corynebacterium spp. diminish S. aureus virulence. We tested this by assaying for changes in S. aureus gene expression during in vitro mono- versus coculture with Corynebacterium striatum, a common skin and nasal commensal. We observed a broad shift in S. aureus gene transcription during in vitro growth with C. striatum, including increased transcription of genes known to exhibit increased expression during human nasal colonization and decreased transcription of virulence genes. S. aureus uses several regulatory pathways to transition between commensal and pathogenic states. One of these, the quorum signal accessory gene regulator (agr) system, was strongly inhibited in response to Corynebacterium spp. Phenotypically, S. aureus exposed to C. striatum exhibited increased adhesion to epithelial cells, reflecting a commensal state, and decreased hemolysin activity, reflecting an attenuation of virulence. Consistent with this, S. aureus displayed diminished fitness in experimental in vivo coinfection with C. striatum when compared to monoinfection. These data support a model in which S. aureus shifts from virulence toward a commensal state when exposed to commensal Corynebacterium species.

  8. Staphylococcus aureus Shifts toward Commensalism in Response to Corynebacterium Species.

    Science.gov (United States)

    Ramsey, Matthew M; Freire, Marcelo O; Gabrilska, Rebecca A; Rumbaugh, Kendra P; Lemon, Katherine P

    2016-01-01

    Staphylococcus aureus-human interactions result in a continuum of outcomes from commensalism to pathogenesis. S. aureus is a clinically important pathogen that asymptomatically colonizes ~25% of humans as a member of the nostril and skin microbiota, where it resides with other bacteria including commensal Corynebacterium species. Commensal Corynebacterium spp. are also positively correlated with S. aureus in chronic polymicrobial diabetic foot infections, distinct from acute monomicrobial S. aureus infections. Recent work by our lab and others indicates that microbe-microbe interactions between S. aureus and human skin/nasal commensals, including Corynebacterium species, affect S. aureus behavior and fitness. Thus, we hypothesized that S. aureus interactions with Corynebacterium spp. diminish S. aureus virulence. We tested this by assaying for changes in S. aureus gene expression during in vitro mono- versus coculture with Corynebacterium striatum, a common skin and nasal commensal. We observed a broad shift in S. aureus gene transcription during in vitro growth with C. striatum, including increased transcription of genes known to exhibit increased expression during human nasal colonization and decreased transcription of virulence genes. S. aureus uses several regulatory pathways to transition between commensal and pathogenic states. One of these, the quorum signal accessory gene regulator (agr) system, was strongly inhibited in response to Corynebacterium spp. Phenotypically, S. aureus exposed to C. striatum exhibited increased adhesion to epithelial cells, reflecting a commensal state, and decreased hemolysin activity, reflecting an attenuation of virulence. Consistent with this, S. aureus displayed diminished fitness in experimental in vivo coinfection with C. striatum when compared to monoinfection. These data support a model in which S. aureus shifts from virulence toward a commensal state when exposed to commensal Corynebacterium species. PMID:27582729

  9. Pleurodesis with doxycycline or Corynebacterium parvum in malignant pleural effusion.

    Science.gov (United States)

    Salomaa, E R; Pulkki, K; Helenius, H

    1995-01-01

    Pleurodesis with doxycycline (100 mg and 600 mg) and Corynebacterium parvum (1 mg and 7 mg) were compared in 41 patients with malignant effusion. To evaluate the mechanisms, pleural fluid pH, leukocytes, granulocytes, interleukin-6 (IL-6) and serum IL-6, as well as C-reactive protein (CRP) were measured before and on 2 consecutive days after treatment. Corynebacterium parvum produced a greater acute-phase response measured with fever, serum CRP and IL-6 than doxycycline. However, no change in pleural fluid IL-6 was demonstrated. Among the 35 assessed patients, 26 had objective response, similar in all four treatment groups. Side-effects were more common with Corynebacterium parvum. Based on this preliminary study we conclude that doxycycline, even in low doses, is a highly effective and well tolerated agent for palliative treatment of malignant pleural effusion. As the responses were similar despite different inflammatory reactions, the two agents probably induce pleural obliteration through different mechanisms. PMID:7865226

  10. PRODUKSI L-LISIN OLEH GALUR MUTAN Corynebacterium glutamicum DENGAN MEMANFAATKAN MOLASE L-LYSINE PRODUCTION BY MUTANT STRAINS OF Corynebacterium glutamicum USING MOLASSES

    OpenAIRE

    Budiatman Satiawihardja; Erliza Noor*; Ahmad Haryo Oktamto 3

    2000-01-01

    The objective of this study was to obtain fermentation process for producing l-lysine using two mutans of Corynebacterium glutamicum. The process used a low cost natural raw material molasses as a carbon source which underwent a special treatment before its application for fermentation medium. Strain Corynebacterium glutamicum ATCC 21543 produced higher L-lysine as compared to Corynebacterium glutamicum ATCC 21513. ATCC strain 21543 produced using a 10.98 g/l L-lysine modified basal ...

  11. Experimental transmission of Corynebacterium pseudotuberculosis in horses by house flies

    Science.gov (United States)

    The route of infection of pigeon fever remains undetermined. The purpose of this study was to investigate house flies (Musca domestica L.) as vectors of Corynebacterium pseudotuberculosis in horses. Eight ponies were used in a randomized, controlled, blinded experimental study. Ten wounds were creat...

  12. Early prosthetic valve endocarditis caused by Corynebacterium kroppenstedtii.

    Science.gov (United States)

    Hagemann, Jürgen Benjamin; Essig, Andreas; Herrmann, Manuel; Liebold, Andreas; Quader, Mohamed Abo

    2015-12-01

    Corynebacterium (C.) kroppenstedtii is a rarely detected agent of bacterial infections in humans. Here, we describe the first case of prosthetic valve endocarditis caused by C. kroppenstedtii. Application of molecular methods using surgically excised valve tissue was a cornerstone for the establishment of the microbiological diagnosis, which is crucial for targeted antimicrobial treatment.

  13. Genome sequence of the marine bacterium Corynebacterium maris type strain Coryn-1T (= DSM 45190T)

    OpenAIRE

    Schaffert, Lena; Albersmeier, Andreas; Bednarz, Hanna; Niehaus, Karsten; Kalinowski, Jörn; Rückert, Christian

    2013-01-01

    Corynebacterium maris Coryn-1T Ben-Dov et al. 2009 is a member of the genus Corynebacterium which contains Gram-positive, non-spore forming bacteria with a high G+C content. C. maris was isolated from the mucus of the Scleractinian coral Fungia granulosa and belongs to the aerobic and non-haemolytic corynebacteria. It displays tolerance to salts (up to 10%) and is related to the soil bacterium Corynebacterium halotolerans . As this is a type strain in a subgroup of Corynebacterium without com...

  14. Development of Fatty Acid-Producing Corynebacterium glutamicum Strains

    OpenAIRE

    Takeno, Seiki; Takasaki, Manami; Urabayashi, Akinobu; Mimura, Akinori; Muramatsu, Tetsuhiro; Mitsuhashi, Satoshi; Ikeda, Masato

    2013-01-01

    To date, no information has been made available on the genetic traits that lead to increased carbon flow into the fatty acid biosynthetic pathway of Corynebacterium glutamicum. To develop basic technologies for engineering, we employed an approach that begins by isolating a fatty acid-secreting mutant without depending on mutagenic treatment. This was followed by genome analysis to characterize its genetic background. The selection of spontaneous mutants resistant to the palmitic acid ester s...

  15. Corynebacterium glutamicum Metabolic Engineering with CRISPR Interference (CRISPRi)

    OpenAIRE

    Cleto, Sara; Jensen, Jaide VK; Wendisch, Volker F.; Lu, Timothy K.

    2016-01-01

    Corynebacterium glutamicum is an important organism for the industrial production of amino acids. Metabolic pathways in this organism are usually engineered by conventional methods such as homologous recombination, which depends on rare double-crossover events. To facilitate the mapping of gene expression levels to metabolic outputs, we applied CRISPR interference (CRISPRi) technology using deactivated Cas9 (dCas9) to repress genes in C. glutamicum. We then determined the effects of target re...

  16. Further experience of Corynebacterium parvum in malignant pleural effusion.

    OpenAIRE

    McLeod, D T; Calverley, P M; Millar, J W; Horne, N. W.

    1985-01-01

    The outcome of drug induced pleurodesis has been evaluated in a non-randomised retrospective study of 67 patients with recurrent malignant pleural effusions treated during 1976-83. Fourteen died within 30 days of treatment. Of the remaining 53 patients, 26 studied during 1976-80 were treated with intrapleural mustine hydrochloride, while 27 studied during 1978-83 were treated with intrapleural Corynebacterium parvum. There was no significant difference between the groups in age, sex, or tumou...

  17. Histidine biosynthesis, its regulation and biotechnological application in Corynebacterium glutamicum

    OpenAIRE

    Kulis-Horn, Robert K; Persicke, Marcus; Kalinowski, Jörn

    2013-01-01

    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...

  18. Interdigital foot infections: Corynebacterium minutissimum and agents of superficial mycoses

    Directory of Open Access Journals (Sweden)

    Fatma Mutlu Sariguzel

    2014-09-01

    Full Text Available Interdigital foot infections are mostly caused initially by dermatophytes, yeasts and less frequently by bacteria. Erythrasma caused by Corynebacterium minutissimum can be confused with superficial mycoses. The aim of the study was to determine the prevalence of the etiologic agents of superficial mycoses and the frequency of Corynebacterium minutissimum in interdigital foot infections. All the samples obtained from the 121 patients with interdigital foot infections were examined directly with the use of 20% potassium hydroxide mounts and Gram stain under the microscope and cultured on Sabouraud's dextrose agar plates. In identification of superficial mycoses, the rate was found to be 14% with the cultural method and 14% with direct microscopic examination. Using a combination of direct microscopic examination and culture, a 33.8% ratio was achieved. In the culture of these samples, the most isolated factor was Trichophyton rubrum (33.7%. In 24 of the patients (19.8% Corynebacterium minutissimum was detected by Gram staining, in 6 of these patients Trichophyton rubrum was found, Trichophyton mentagrophytes was found in 2 and Trichosporon spp. was found in 1. The examination of interdigital foot lesions in the laboratory, the coexistence of erythrasma with dermatophytes and yeast should be considered.

  19. Interdigital foot infections: Corynebacterium minutissimum and agents of superficial mycoses

    Science.gov (United States)

    Sariguzel, Fatma Mutlu; Koc, A. Nedret; Yagmur, Gülhan; Berk, Elife

    2014-01-01

    Interdigital foot infections are mostly caused initially by dermatophytes, yeasts and less frequently by bacteria. Erythrasma caused by Corynebacterium minutissimum can be confused with superficial mycoses. The aim of the study was to determine the prevalence of the etiologic agents of superficial mycoses and the frequency of Corynebacterium minutissimum in interdigital foot infections. All the samples obtained from the 121 patients with interdigital foot infections were examined directly with the use of 20% potassium hydroxide mounts and Gram stain under the microscope and cultured on Sabouraud’s dextrose agar plates. In identification of superficial mycoses, the rate was found to be 14% with the cultural method and 14% with direct microscopic examination. Using a combination of direct microscopic examination and culture, a 33.8% ratio was achieved. In the culture of these samples, the most isolated factor was Trichophyton rubrum (33.7%). In 24 of the patients (19.8%) Corynebacterium minutissimum was detected by Gram staining, in 6 of these patients Trichophyton rubrum was found, Trichophyton mentagrophytes was found in 2 and Trichosporon spp. was found in 1. The examination of interdigital foot lesions in the laboratory, the coexistence of erythrasma with dermatophytes and yeast should be considered. PMID:25477907

  20. Immunological characterization of diphtheria toxin recovered from Corynebacterium pseudotuberculosis.

    Science.gov (United States)

    Selim, Salha Abdelkareem; Mohamed, Farida Hessain; Hessain, Ashgan Mohamed; Moussa, Ihab Mohamed

    2016-03-01

    Diphtheria toxin (DT) is a potent toxin produced by the so-called diphtheria group which includes Corynebacterium diphtheriae (C. diphtheriae), Corynebacterium ulcerans (C. ulcerans), and Corynebacterium pseudotuberculosis (C. pseudotuberculosis). The present investigation is aimed to study in detail the production of DT by C. pseudotuberculosis. Twenty isolates were obtained from sheep diseased with caseous lymphadenitis (CLA) and twenty-six isolates were obtained from 26 buffaloes diseased with oedematous skin disease (OSD). All isolates were identified by standard microbiological and DT production was assayed serologically by modified Elek test and immunoblotting. All sheep isolates were nitrate negative, failed to hydrolyze starch and could not produce DT, while all buffalo isolates (biotype II) revealed positive results and a specific band of 62 kDa, specific to DT, was resulted in all concentrated cell fractions (CF), but was absent from non-toxigenic biotype I isolates. At the same time, another band of 31 kDa specific to the PLD gene was obtained with all isolates of biotype I and II. Moreover, all isolates showed positive synergistic hemolytic activity and antagonistic hemolysis with β-hemolytic Staphylococci. The obtained results also indicated that C. pseudotuberculosis could be classified into two strains; non-toxigenic biotype I strain, which failed to produce DT as well as being negative to nitrate and starch hydrolysis, and toxigenic biotype II strain, which can reduce nitrate, hydrolyze starch as well as produce DT. PMID:26981011

  1. Corynebacterium accolens Isolated from Breast Abscess: Possible Association with Granulomatous Mastitis▿

    OpenAIRE

    Ang, Lei M. N.; Brown, Hamish

    2007-01-01

    Corynebacterium accolens is rarely isolated as a human pathogen. We describe here a case of C. accolens isolated from a breast abscess in a patient previously diagnosed with granulomatous mastitis. The possible association of Corynebacterium accolens and granulomatous mastitis in this patient is discussed.

  2. Functional characterization of a vanillin dehydrogenase in Corynebacterium glutamicum

    OpenAIRE

    Ding, Wei; Si, Meiru; Zhang, Weipeng; Zhang, Yaoling; Chen, Can; Zhang, Lei; Lu, Zhiqiang; Chen, Shaolin; Shen, Xihui

    2015-01-01

    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 ~51kDa, whereas the apparent native Mr values revealed by gel filtration chromatography were 49.5, 92.3, 159.0 and 199.2kDa, indicating the presence of dimeric, trimeric and tetrameric forms. Moreover, the enzyme showed its highest level of activity towar...

  3. Corynebacterium pseudotuberculosis RNA-seq data from abiotic stresses

    OpenAIRE

    Pablo H.C.G. de Sá; Veras, Adonney A. O.; Carneiro, Adriana R.; Baraúna, Rafael A.; Luís C. Guimarães; Pinheiro, Kenny C.; Pinto, Anne C.; Soares, Siomar C.; Schneider, Maria P. C.; Vasco Azevedo; Artur Silva; Ramos, Rommel T. J.

    2015-01-01

    Corynebacterium pseudotuberculosis causes significant loss to goat and sheep farmers because it is the causal agent of the infectious disease caseous lymphadenitis, which may lead to outcomes ranging from skin injury to animal death (Ruiz et al., 2011) [1]. This bacterium was grown under osmotic (2 M), acid (pH) and heat (50 °C) stress and under control (Normal-BHI brain heart infusion) conditions, which simulate the conditions faced by the bacteria during the infectious process. Subsequently...

  4. Functional characterization of a vanillin dehydrogenase in Corynebacterium glutamicum.

    Science.gov (United States)

    Ding, Wei; Si, Meiru; Zhang, Weipeng; Zhang, Yaoling; Chen, Can; Zhang, Lei; Lu, Zhiqiang; Chen, Shaolin; Shen, Xihui

    2015-01-27

    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 vanillin at pH 7.0 and 30°C, and interestingly, it could utilize NAD(+) and NADP(+) as coenzymes with similar efficiency and showed no obvious difference toward NAD(+) and NADP(+). In addition to vanillin, this enzyme exhibited catalytic activity toward a broad range of substrates, including p-hydroxybenzaldehyde, 3,4-dihydroxybenzaldehyde, o-phthaldialdehyde, cinnamaldehyde, syringaldehyde and benzaldehyde. Conserved catalytic residues or putative cofactor interactive sites were identified based on sequence alignment and comparison with previous studies, and the function of selected residues were verified by site-directed mutagenesis analysis. Finally, the vdh deletion mutant partially lost its ability to grow on vanillin, indicating the presence of alternative VDH(s) in Corynebacterium glutamicum. Taken together, this study contributes to understanding the VDH diversity from bacteria and the aromatic metabolism pathways in C. glutamicum.

  5. A microbiological and clinical review on Corynebacterium kroppenstedtii.

    Science.gov (United States)

    Tauch, Andreas; Fernández-Natal, Isabel; Soriano, Francisco

    2016-07-01

    The genus Corynebacterium represents a taxon of Gram-positive bacteria with a high G+C content in the genomic DNA. Corynebacterium kroppenstedtii is an unusual member of this taxon as it lacks the characteristic mycolic acids in the cell envelope. Genome sequence analysis of the C. kroppenstedtii type strain has revealed a lipophilic (lipid-requiring) lifestyle and a remarkable repertoire of carbohydrate uptake and utilization systems. Clinical isolates of C. kroppenstedtii have been obtained almost exclusively from female patients and mainly from breast abscesses and cases of granulomatous mastitis. However, the role of C. kroppenstedtii in breast pathologies remains unclear. This review provides a comprehensive overview of the taxonomy, microbiology, and microbiological identification of C. kroppenstedtii, including polyphasic phenotypic approaches, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the use of 16S rRNA gene sequencing. A clinical review presents reported cases, various antimicrobial treatments, antibiotic susceptibility assays, and antibiotic resistance genes detected during genome sequencing. C. kroppenstedtii must be considered a potential opportunistic human pathogen and should be identified accurately in clinical laboratories. PMID:27155209

  6. Genome sequence of the marine bacterium Corynebacterium maris type strain Coryn-1(T) (= DSM 45190(T)).

    Science.gov (United States)

    Schaffert, Lena; Albersmeier, Andreas; Bednarz, Hanna; Niehaus, Karsten; Kalinowski, Jörn; Rückert, Christian

    2013-07-30

    Corynebacterium maris Coryn-1(T) Ben-Dov et al. 2009 is a member of the genus Corynebacterium which contains Gram-positive, non-spore forming bacteria with a high G+C content. C. maris was isolated from the mucus of the Scleractinian coral Fungia granulosa and belongs to the aerobic and non-haemolytic corynebacteria. It displays tolerance to salts (up to 10%) and is related to the soil bacterium Corynebacterium halotolerans. As this is a type strain in a subgroup of Corynebacterium without complete genome sequences, this project, describing the 2.78 Mbp long chromosome and the 45.97 kbp plasmid pCmaris1, with their 2,584 protein-coding and 67 RNA genes, will aid the G enomic E ncyclopedia of Bacteria and Archaea project. PMID:24501635

  7. Experimental transmission of Corynebacterium pseudotuberculosis biovar equi in horses by house flies

    Science.gov (United States)

    The route of Corynebacterium pseudotuberculosis infection in horses remains undetermined, but transmission by insects is suspected. Scientists from CMAVE and Auburn University investigated house flies (Musca domestica L.) as possible vectors. Three ponies were directly inoculated with C. pseudotuber...

  8. Corynebacterium ulcerans in an Immunocompromised Patient with Diphtheria and Her Dog

    OpenAIRE

    Lartigue, Marie-Frédérique; Monnet, Xavier; Le Flèche, Anne; Grimont, Patrick A. D.; Benet, Jean-Jacques; Durrbach, Antoine; Fabre, Monique; Nordmann, Patrice

    2005-01-01

    Corynebacterium ulcerans causes zoonotic infections, such as diphtheria and extrapharyngeal infections. We report here the first case of a diphtheria-like illness caused by C. ulcerans in France and transmitted likely by a dog to an immunocompromised woman.

  9. Corynebacterium glutamicum Metabolic Engineering with CRISPR Interference (CRISPRi).

    Science.gov (United States)

    Cleto, Sara; Jensen, Jaide Vk; Wendisch, Volker F; Lu, Timothy K

    2016-05-20

    Corynebacterium glutamicum is an important organism for the industrial production of amino acids. Metabolic pathways in this organism are usually engineered by conventional methods such as homologous recombination, which depends on rare double-crossover events. To facilitate the mapping of gene expression levels to metabolic outputs, we applied CRISPR interference (CRISPRi) technology using deactivated Cas9 (dCas9) to repress genes in C. glutamicum. We then determined the effects of target repression on amino acid titers. Single-guide RNAs directing dCas9 to specific targets reduced expression of pgi and pck up to 98%, and of pyk up to 97%, resulting in titer enhancement ratios of l-lysine and l-glutamate production comparable to levels achieved by gene deletion. This approach for C. glutamicum metabolic engineering, which only requires 3 days, indicates that CRISPRi can be used for quick and efficient metabolic pathway remodeling without the need for gene deletions or mutations and subsequent selection.

  10. Corynebacterium pseudotuberculosis RNA-seq data from abiotic stresses.

    Science.gov (United States)

    de Sá, Pablo H C G; Veras, Adonney A O; Carneiro, Adriana R; Barúna, Rafael A; Guimarães, Luís C; Pinheiro, Kenny C; Pinto, Anne C; Soares, Siomar C; Schneider, Maria P C; Azevedo, Vasco; Silva, Artur; Ramos, Rommel T J

    2015-12-01

    Corynebacterium pseudotuberculosis causes significant loss to goat and sheep farmers because it is the causal agent of the infectious disease caseous lymphadenitis, which may lead to outcomes ranging from skin injury to animal death (Ruiz et al., 2011) [1]. This bacterium was grown under osmotic (2 M), acid (pH) and heat (50 °C) stress and under control (Normal-BHI brain heart infusion) conditions, which simulate the conditions faced by the bacteria during the infectious process. Subsequently, cDNA of each condition was sequenced by the SOLiD3 Plus platform using the RNA-Seq technique [2], [3], [4]. The data produced was processed to evaluate the differential gene expression, which is helpful to understand the adaptation mechanisms during the infection in the host. The sequencing data of all conditions are available in the European Bioinformatics Institute (EBI) repository under accession number E-MTAB-2017. PMID:26702428

  11. Corynebacterium pseudotuberculosis RNA-seq data from abiotic stresses

    Directory of Open Access Journals (Sweden)

    Pablo H.C.G. de Sá

    2015-12-01

    Full Text Available Corynebacterium pseudotuberculosis causes significant loss to goat and sheep farmers because it is the causal agent of the infectious disease caseous lymphadenitis, which may lead to outcomes ranging from skin injury to animal death (Ruiz et al., 2011 [1]. This bacterium was grown under osmotic (2 M, acid (pH and heat (50 °C stress and under control (Normal-BHI brain heart infusion conditions, which simulate the conditions faced by the bacteria during the infectious process. Subsequently, cDNA of each condition was sequenced by the SOLiD3 Plus platform using the RNA-Seq technique [2–4]. The data produced was processed to evaluate the differential gene expression, which is helpful to understand the adaptation mechanisms during the infection in the host. The sequencing data of all conditions are available in the European Bioinformatics Institute (EBI repository under accession number E-MTAB-2017.

  12. CoryneBase: Corynebacterium genomic resources and analysis tools at your fingertips.

    Directory of Open Access Journals (Sweden)

    Hamed Heydari

    Full Text Available Corynebacteria are used for a wide variety of industrial purposes but some species are associated with human diseases. With increasing number of corynebacterial genomes having been sequenced, comparative analysis of these strains may provide better understanding of their biology, phylogeny, virulence and taxonomy that may lead to the discoveries of beneficial industrial strains or contribute to better management of diseases. To facilitate the ongoing research of corynebacteria, a specialized central repository and analysis platform for the corynebacterial research community is needed to host the fast-growing amount of genomic data and facilitate the analysis of these data. Here we present CoryneBase, a genomic database for Corynebacterium with diverse functionality for the analysis of genomes aimed to provide: (1 annotated genome sequences of Corynebacterium where 165,918 coding sequences and 4,180 RNAs can be found in 27 species; (2 access to comprehensive Corynebacterium data through the use of advanced web technologies for interactive web interfaces; and (3 advanced bioinformatic analysis tools consisting of standard BLAST for homology search, VFDB BLAST for sequence homology search against the Virulence Factor Database (VFDB, Pairwise Genome Comparison (PGC tool for comparative genomic analysis, and a newly designed Pathogenomics Profiling Tool (PathoProT for comparative pathogenomic analysis. CoryneBase offers the access of a range of Corynebacterium genomic resources as well as analysis tools for comparative genomics and pathogenomics. It is publicly available at http://corynebacterium.um.edu.my/.

  13. Structural basis for cytokinin production by LOG from Corynebacterium glutamicum.

    Science.gov (United States)

    Seo, Hogyun; Kim, Sangwoo; Sagong, Hye-Young; Son, Hyeoncheol Francis; Jin, Kyeong Sik; Kim, Il-Kwon; Kim, Kyung-Jin

    2016-08-10

    "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 as a dimer and shows an overall structure similar to other known LOGs, such as LOGs from Arabidopsis thaliana (AtLOG), Claviceps purpurea (CpLOG), and Mycobacterium marinum (MmLOG). Cg2612 also contains a "PGGXGTXXE" motif that contributes to the formation of an active site similar to other LOGs. Moreover, biochemical studies on Cg2612 revealed that the protein has phosphoribohydrolase activity but not LDC activity. Based on these structural and biochemical studies, we propose that Cg2612 is not an LDC family enzyme, but instead belongs to the LOG family. In addition, the prenyl-binding site of Cg2612 (CgLOG) comprised residues identical to those seen in AtLOG and CpLOG, albeit dissimilar to those in MmLOG. The work provides structural and functional implications for LOG-like proteins from other microorganisms.

  14. A combined approach for comparative exoproteome analysis of Corynebacterium pseudotuberculosis

    Directory of Open Access Journals (Sweden)

    Scrivens James H

    2011-01-01

    Full Text Available Abstract Background Bacterial exported proteins represent key components of the host-pathogen interplay. Hence, we sought to implement a combined approach for characterizing the entire exoproteome of the pathogenic bacterium Corynebacterium pseudotuberculosis, the etiological agent of caseous lymphadenitis (CLA in sheep and goats. Results An optimized protocol of three-phase partitioning (TPP was used to obtain the C. pseudotuberculosis exoproteins, and a newly introduced method of data-independent MS acquisition (LC-MSE was employed for protein identification and label-free quantification. Additionally, the recently developed tool SurfG+ was used for in silico prediction of sub-cellular localization of the identified proteins. In total, 93 different extracellular proteins of C. pseudotuberculosis were identified with high confidence by this strategy; 44 proteins were commonly identified in two different strains, isolated from distinct hosts, then composing a core C. pseudotuberculosis exoproteome. Analysis with the SurfG+ tool showed that more than 75% (70/93 of the identified proteins could be predicted as containing signals for active exportation. Moreover, evidence could be found for probable non-classical export of most of the remaining proteins. Conclusions Comparative analyses of the exoproteomes of two C. pseudotuberculosis strains, in addition to comparison with other experimentally determined corynebacterial exoproteomes, were helpful to gain novel insights into the contribution of the exported proteins in the virulence of this bacterium. The results presented here compose the most comprehensive coverage of the exoproteome of a corynebacterial species so far.

  15. Intrapleural Corynebacterium parvum for recurrent malignant pleural effusions.

    Science.gov (United States)

    Foresti, V

    1995-01-01

    Twenty-two consecutive patients with malignant pleural effusions (MPE) were treated with intrapleural Corynebacterium parvum (CBP) associated with parenteral methylprednisolone (MP) to determine its effectiveness and the frequency and nature of adverse reactions. After thoracentesis, 7 mg of CBP (Coparvax Wellcome) in 20 ml of saline were injected into the pleural cavity. On the day of treatment, the patients were given 1 mg/kg i.m. of MP 30 min before thoracentesis. The effectiveness of pleurodesis was assessed as follows: (1) complete response (CR; total resolution of pleural effusion after 3 injections of CBP at the most); (2) partial response (PR; formation of asymptomatic loculated effusion). In 5 patients leukocytes, lymphocytes and monocytes were determined in pleural fluid (PF) and in blood (B) collected before and 7 days after CBP treatment. Two patients were unevaluable. Of 20 evaluable patients, 18 (90%) had a CR and 2 patients (10%) had a PR. Eleven of 22 patients (50%) had a fever. Three patients had prolonged and/or high fever. Seven of 22 patients (32%) had mild chest pain. None of the patients presented other side effects. Twelve of 21 patients (57.1%) had a PF pH > or = 7.30; 2 of these died a few days after the treatment, and 10 had favorable responses. The other 9 patients had a PF pH treatment. Our study confirms that intrapleural CBP is an effective and simple method to control MPE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7716350

  16. Biofilm production by multiresistant Corynebacterium striatum associated with nosocomial outbreak

    Science.gov (United States)

    de Souza, Cassius; Faria, Yuri Vieira; Sant’Anna, Lincoln de Oliveira; Viana, Vanilda Gonçalves; Seabra, Sérgio Henrique; de Souza, Mônica Cristina; Vieira, Verônica Viana; Hirata, Raphael; Moreira, Lílian de Oliveira; de Mattos-Guaraldi, Ana Luíza

    2015-01-01

    Corynebacterium striatum is a potentially pathogenic microorganism that causes nosocomial outbreaks. However, little is known about its virulence factors that may contribute to healthcare-associated infections (HAIs). We investigated the biofilm production on abiotic surfaces of multidrug-resistant (MDR) and multidrug-susceptible (MDS) strains of C. striatum of pulsed-field gel electrophoresis types I-MDR, II-MDR, III-MDS and IV-MDS isolated during a nosocomial outbreak in Rio de Janeiro, Brazil. The results showed that C. striatum was able to adhere to hydrophilic and hydrophobic abiotic surfaces. The C. striatum 1987/I-MDR strain, predominantly isolated from patients undergoing endotracheal intubation procedures, showed the greatest ability to adhere to all surfaces. C. striatum bound fibrinogen to its surface, which contributed to biofilm formation. Scanning electron microscopy showed the production of mature biofilms on polyurethane catheters by all pulsotypes. In conclusion, biofilm production may contribute to the establishment of HAIs caused by C. striatum. PMID:25946249

  17. Corynebacterium glutamicum Metabolic Engineering with CRISPR Interference (CRISPRi).

    Science.gov (United States)

    Cleto, Sara; Jensen, Jaide Vk; Wendisch, Volker F; Lu, Timothy K

    2016-05-20

    Corynebacterium glutamicum is an important organism for the industrial production of amino acids. Metabolic pathways in this organism are usually engineered by conventional methods such as homologous recombination, which depends on rare double-crossover events. To facilitate the mapping of gene expression levels to metabolic outputs, we applied CRISPR interference (CRISPRi) technology using deactivated Cas9 (dCas9) to repress genes in C. glutamicum. We then determined the effects of target repression on amino acid titers. Single-guide RNAs directing dCas9 to specific targets reduced expression of pgi and pck up to 98%, and of pyk up to 97%, resulting in titer enhancement ratios of l-lysine and l-glutamate production comparable to levels achieved by gene deletion. This approach for C. glutamicum metabolic engineering, which only requires 3 days, indicates that CRISPRi can be used for quick and efficient metabolic pathway remodeling without the need for gene deletions or mutations and subsequent selection. PMID:26829286

  18. Analysis and Engineering of Metabolic Pathway Fluxes in Corynebacterium glutamicum

    Science.gov (United States)

    Wittmann, Christoph

    The Gram-positive soil bacterium Corynebacterium glutamicum was discovered as a natural overproducer of glutamate about 50 years ago. Linked to the steadily increasing economical importance of this microorganism for production of glutamate and other amino acids, the quest for efficient production strains has been an intense area of research during the past few decades. Efficient production strains were created by applying classical mutagenesis and selection and especially metabolic engineering strategies with the advent of recombinant DNA technology. Hereby experimental and computational approaches have provided fascinating insights into the metabolism of this microorganism and directed strain engineering. Today, C. glutamicum is applied to the industrial production of more than 2 million tons of amino acids per year. The huge achievements in recent years, including the sequencing of the complete genome and efficient post genomic approaches, now provide the basis for a new, fascinating era of research - analysis of metabolic and regulatory properties of C. glutamicum on a global scale towards novel and superior bioprocesses.

  19. Functional characterization of Corynebacterium glutamicum mycothiol S-conjugate amidase.

    Directory of Open Access Journals (Sweden)

    Meiru Si

    Full Text Available The present study focuses on the genetic and biochemical characterization of mycothiol S-conjugate amidase (Mca of Corynebacterium glutamicum. Recombinant C. glutamicum Mca was heterologously expressed in Escherichia coli and purified to apparent homogeneity. The molecular weight of native Mca protein determined by gel filtration chromatography was 35 kDa, indicating that Mca exists as monomers in the purification condition. Mca showed amidase activity with mycothiol S-conjugate of monobromobimane (MSmB in vivo while mca mutant lost the ability to cleave MSmB. In addition, Mca showed limited deacetylase activity with N-acetyl-D-glucosamine (GlcNAc as substrate. Optimum pH for amidase activity was between 7.5 and 8.5, while the highest activity in the presence of Zn2+ confirmed Mca as a zinc metalloprotein. Amino acid residues conserved among Mca family members were located in C. glutamicum Mca and site-directed mutagenesis of these residues indicated that Asp14, Tyr137, His139 and Asp141 were important for activity. The mca deletion mutant showed decreased resistance to antibiotics, alkylating agents, oxidants and heavy metals, and these sensitive phenotypes were recovered in the complementary strain to a great extent. The physiological roles of Mca in resistance to various toxins were further supported by the induced expression of Mca in C. glutamicum under various stress conditions, directly under the control of the stress-responsive extracytoplasmic function-sigma (ECF-σ factor SigH.

  20. Effect of Corynebacterium glutamicum on Livestock Material Burial Treatment.

    Science.gov (United States)

    Kim, Bit-Na; Cho, Ho-Seong; Cha, Yougin; Park, Joon-Kyu; Kim, Geonha; Kim, Yang-Hoon; Min, Jiho

    2016-08-28

    In recent years, foot-and-mouth disease has occurred in all parts of the world. The animals with the disease are buried in the ground; therefore, their concentration could affect ground or groundwater. Moreover, the complete degradation of carcasses is not a certainty, and their disposal is important to prevent humans, livestock, and the environment from being affected with the disease. The treatment of Corynebacterium glutamicum is a feasible method to reduce the risk of carcass decomposition affecting humans or the environment. Therefore, this study aimed to investigate the effect of C. glutamicum on the soil environment with a carcass. The composition of amino acids in the soil treated with C. glutamicum was generally higher than those in the untreated soil. Moreover, the plant root in the soil samples treated with C. glutamicum had 84.0% amino acids relative to the standard value and was similar to that of the control. The results of this study suggest the possibility to reduce the toxicity of a grave land containing animals with this disease. PMID:27160580

  1. Properties of Cassava Starch Modified by Amylomaltase from Corynebacterium glutamicum.

    Science.gov (United States)

    Suriyakul Na Ayudhaya, Pitcha; Pongsawasdi, Piamsook; Laohasongkram, Kalaya; Chaiwanichsiri, Saiwarun

    2016-06-01

    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 apparent amylose content and retrogradation enthalpy were lower, whereas the retrogradation temperatures, freeze-thaw stability, and thermo-reversibility were higher than those of the native cassava starch. However, when amylomaltase content was increased to 20 units of amylomaltase/g starch and for 24 h, the modified starch showed an improvement in the thermo-reversibility property. When used in panna cotta, the gel strength of the sample using the 20 units/24 h modified cassava starch was similar to that of using gelatin. PMID:27105125

  2. Diagnostic and experimental study of Corynebacterium renale isolated from urinary tract infection of cattle

    Directory of Open Access Journals (Sweden)

    S. A. Hussein

    2011-01-01

    Full Text Available The study includes isolation and identification of Corynebacterium renale from urine of cow apparently suffering from urinary tract infection. C. renale represent highest isolate 49. 99% followed by Corynebacterium pyogenes 24.24% from the total number of Corynebacterium 74.23%. on the other hand Staphylococcus saprophyticus also isolated from urine samples 25.75%. Since C. renale was isolated at highest rate we studied its pathogenesis via inoculation of isolate intraperitoneally into white Swiss mice. Results showed that C. renale type I has ability to produce kidney damage after 48 hr. post inoculation revealed embolic glomeruler nephritis with less number of C. renale, also there is infiltration of polymorphnuclear inflammatory cell and nephrosis, in addition to vacular degeneration, coagulative necrosis with blood vessel congestion in liver tissue.

  3. Dicty_cDB: Contig-U16321-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available :none) Corynebacterium aurimucosum ATC... 50 2e-04 BC029705_1( BC029705 |pid:none) Mus musculus suprabasin...coccus geothermalis DSM 11... 43 0.027 (A6QQF6) RecName: Full=Suprabasin; Flags:

  4. METODE CEPAT EKSTRAKSI DNA Corynebacterium diphtheriae UNTUK PEMERIKSAAN PCR

    Directory of Open Access Journals (Sweden)

    Sunarno Sunarno

    2014-10-01

    Full Text Available AbstractDiagnosis of diphtheria caused byCorynebacterium diphtheriaeshould be done immediately since delay of therapy may cause 20-fold increase rate of death. One method of rapid diagnostic to identify diphtheria is by using polymerase chain reaction (PCR. The fundamental issue of this method depends on the DNA, either its quality or quantity. The simple DNA extraction method, which is using mechanical/physical principles with a little of chemical reagents (such as boiling method and the use of sodium hydroxide (NAOH, will have some benefits, such as easy to be performed, low cost, fast, and environmentally friendly. This study aimed to evaluate effectivity and efficiency of boiling method with NaOH to extract DNA of C. diphtheriae compared to the use of a commercial diagnostic kit for PCR assay. We used C. diphtheriae toxygenic(NCTC 10648 isolates, which are grown in blood agar plates. We then prepared the suspensions of cell/colony in aquadest with several dilutions. Each dilution was extracted using boiling method, NaOH and controlled with the use of a commercial diagnostic kit (QiAmp DNA Minikit. The results were evaluated quantitatively with spectrophotometer and qualitatively with gel electrophoresis. The results showed that the extracted DNA from boiling method with NaOH has an adequate quality and quantity for PCR assay (up to 9 CFU/uL cell/reaction. Therefore, it can be summarized that boiling method with NaOH is effective and efficient to be applied in PCR assay forC. diphtheriae.Key words: boiling extraction method, NaOH, C.diphtheriae, PCRAbstrakKematian kasus difteri yang disebabkan oleh Corynebacterium diphtheriaedapat meningkat 20 kali lipat karena keterlambatan pengobatan sehingga penegakan diagnosis harus dilakukan sesegera mungkin. Salah satu metode diagnostik yang cukup cepat untuk mendeteksi penyakit difteri adalah pemeriksaan polymerase chain reaction(PCR. Keberhasilan pemeriksaan PCR dipengaruhi oleh kualitas dan kuantitas

  5. Activity of disinfectants and biofilm production of Corynebacterium pseudotuberculosis

    Directory of Open Access Journals (Sweden)

    Maria da C.A. Sá

    2013-11-01

    Full Text Available To verify the occurrence of caseous lymphadenitis in sheep and goats on farms of Pernambuco, Brazil, and in animals slaughtered in two Brazilian cities (Petrolina/PE and Juazeiro/BA, and to characterize the susceptibility profile of Corynebacterium pseudotuberculosis to disinfectants and antimicrobials, and its relationship with biofilm production were the objectives of this study. 398 samples were tested for sensitivity to antimicrobial drugs, disinfectants, and biofilm production. Among the 108 samples collected on the properties, 75% were positive for C. pseudotuberculosis. Slaughterhouse samples indicated an occurrence of caseous lymphadenitis in 15.66% and 6.31% for animals slaughtered in Petrolina and Juazeiro respectively. With respect to antimicrobials, the sensitivity obtained was 100% for florfenicol and tetracycline; 99.25% for enrofloxacin, ciprofloxacin and lincomycin; 98.99% for cephalothin; 98.74% for norfloxacin and sulfazotrim; 97.74% for gentamicin; 94.22% for ampicillin; 91.71% for amoxicillin; 91.21% for penicillin G; 89.19% for neomycin and 0% for novobiocin. In analyzes with disinfectants, the efficiency for chlorhexidine was 100%, 97.20% for quaternary ammonium, 87.40% for chlorine and 84.40% for iodine. 75% of the isolates were weak or non-biofilm producers. For the consolidated biofilm, found that iodine decreased biofilm formation in 13 isolates and quaternary ammonia in 11 isolates. The reduction of the biofilm formation was observed for iodine and quaternary ammonium in consolidated biofilm formation in 33% and 28% of the isolates, respectively. The results of this study highlight the importance of establishing measures to prevent and control the disease.

  6. Detection of Differences in the Nucleotide and Amino Acid Sequences of Diphtheria Toxin from Corynebacterium diphtheriae and Corynebacterium ulcerans Causing Extrapharyngeal Infections

    OpenAIRE

    Sing, Andreas; Hogardt, Michael; Bierschenk, Suse; Heesemann, Jürgen

    2003-01-01

    While Corynebacterium ulcerans can mimic classical diphtheria, extrapharyngeal infections are extremely rare. Sequencing of the diphtheria toxin (DT)-encoding tox gene of two C. ulcerans isolates from extrapharyngeal infections revealed differences from C. diphtheriae DT sequences, mainly in the translocation and receptor-binding domains. C. ulcerans supernatants were much less potent than supernatant from C. diphtheriae. A C. ulcerans DT-specific PCR is described below.

  7. Draft Genome Sequence of Corynebacterium ulcerans Strain 04-3911, Isolated from Humans

    Science.gov (United States)

    Viana, Marcus V. C.; Benevides, Leandro J.; Mariano, Diego C. B.; Veras, Adooney A. O.; Sá, Pablo H. C.; Rocha, Flávia S.; Vilas Boas, Priscilla C. B.; Soares, Siomar C.; Barbosa, Maria S.; Guiso, Nicole; Badell, Edgar; Carneiro, Adriana R.; Azevedo, Vasco; Ramos, Rommel T. J.

    2016-01-01

    Corynebacterium ulcerans is a pathogenic bacterium infecting wild and domesticated animals; some infection cases in humans have increased throughout the world. The current study describes the draft genome of strain 04-3911, isolated from humans. The draft genome has 2,492,680 bp, 2,143 coding sequences, 12 rRNA genes, and 50 tRNA genes. PMID:27034486

  8. Genomic analysis of a nontoxigenic, invasive Corynebacterium diphtheriae strain from Brazil.

    Science.gov (United States)

    Encinas, Fernando; Marin, Michel A; Ramos, Juliana N; Vieira, Verônica V; Mattos-Guaraldi, Ana Luiza; Vicente, Ana Carolina P

    2015-09-01

    We report the complete genome sequence and analysis of an invasive Corynebacterium diphtheriae strain that caused endocarditis in Rio de Janeiro, Brazil. It was selected for sequencing on the basis of the current relevance of nontoxigenic strains for public health. The genomic information was explored in the context of diversity, plasticity and genetic relatedness with other contemporary strains.

  9. Septic arthritis of a native knee joint due to Corynebacterium striatum.

    Science.gov (United States)

    Westblade, Lars F; Shams, Farah; Duong, Scott; Tariq, Oosman; Bulbin, Alan; Klirsfeld, Dava; Zhen, Wei; Sakaria, Smita; Ford, Bradley A; Burnham, Carey-Ann D; Ginocchio, Christine C

    2014-05-01

    We report a case of septic arthritis of a native knee joint due to Corynebacterium striatum, a rare and unusual cause of septic arthritis of native joints. The isolate was identified by a combination of phenotypic, mass spectrometric, and nucleic acid-based assays and exhibited high-level resistance to most antimicrobials.

  10. Laboratory review of reference strains of Corynebacterium diphtheriae indicates mistyped intermedius strains.

    OpenAIRE

    Coyle, M B; Nowowiejski, D J; Russell, J Q; Groman, N B

    1993-01-01

    All biotyped strains of Corynebacterium diphtheriae from the American Type Culture Collection (ATCC) were compared for morphology and biochemical reactions. Biotypes of all gravis strains and most mitis strains were confirmed, but intermedius strains were found to be misclassified. New lipid-dependent intermedius strains have been deposited with the ATCC.

  11. Pancreatic panniculitis complicated by infection with Corynebacterium tuberculostearicum: A case report

    Directory of Open Access Journals (Sweden)

    S.H. Omland

    2014-01-01

    Full Text Available We present a case of pancreatic panniculitis in a patient with alcohol abuse where Corynebacterium tuberculostearicum was isolated from a pannicular nodule on the crus. The patient was started on linezolid treatment leading to regression of the patient's symptoms. Upon discontinuation of linezolid treatment progression of the skin symptoms progressed.

  12. Trapping and Killing of Candida albicans by Corynebacterium parvum-Activated Livers †

    OpenAIRE

    Sawyer, R T; Moon, R J; Beneke, E S

    1981-01-01

    Corynebacterium parvum vaccination significantly increased the number of leukocytes adherent to hepatic vessels. Perfused C. parvum-treated livers killed significantly more Candida albicans than did livers not treated with C. parvum, an effect reversed by the macrophage inhibitors silica, phenylbutazone, and iodoacetate.

  13. Draft Genome Sequence of Toxigenic Corynebacterium ulcerans Strain 03-8664 Isolated from a Human Throat.

    Science.gov (United States)

    Guimarães, Luis C; Viana, Marcus V C; Benevides, Leandro J; Mariano, Diego C B; Veras, Adonney A O; Sá, Pablo H C; Rocha, Flávia S; Vilas Boas, Priscilla C B; Soares, Siomar C; Barbosa, Maria S; Guiso, Nicole; Badell, Edgar; Azevedo, Vasco; Ramos, Rommel T J; Silva, Artur

    2016-01-01

    Corynebacterium ulcerans is an emergent pathogen infecting wild and domesticated animals worldwide that may serve as reservoirs for zoonotic infections. In this study, we present the draft genome of C. ulcerans strain 03-8664. The draft genome has 2,428,683 bp, 2,262 coding sequences, and 12 rRNA genes.

  14. Complete Sucrose Metabolism Requires Fructose Phosphotransferase Activity in Corynebacterium glutamicum To Ensure Phosphorylation of Liberated Fructose

    OpenAIRE

    Dominguez, H.; Lindley, N. D.

    1996-01-01

    Sucrose uptake by Corynebacterium glutamicum involves a phosphoenolpyruvate-dependent sucrose phosphotransferase (PTS), but in the absence of fructokinase, further metabolism of the liberated fructose requires efflux of the fructose and reassimilation via the fructose PTS. Mutant strains lacking detectable fructose-transporting PTS activity accumulated fructose extracellularly but consumed sucrose at rates comparable to those of the wild-type strain.

  15. Multiplex polymerase chain reaction to identify and determine the toxigenicity of Corynebacterium spp with zoonotic potential and an overview of human and animal infections

    OpenAIRE

    Luciene de Fátima Costa Torres; Dayana Ribeiro; Raphael Hirata Jr; Luis Gustavo Carvalho Pacheco; Monica Cristina de Souza; Louisy Sanches dos Santos; Cíntia Silva dos Santos; Mohammad Salah; Mateus Matiuzzi da Costa; Marcio Garcia Ribeiro; Selim, Salah A; Vasco Ariston de Carvalho Azevedo; Ana Luiza Mattos-Guaraldi

    2013-01-01

    Corynebacterium diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis constitute a group of potentially toxigenic microorganisms that are related to different infectious processes in animal and human hosts. Currently, there is a lack of information on the prevalence of disease caused by these pathogens, which is partially due to a reduction in the frequency of routine laboratory testing. In this study, a multiplex polymerase chain reaction (mPCR) assay that can simultan...

  16. Screening for Corynebacterium diphtheriae and Corynebacterium ulcerans in patients with upper respiratory tract infections 2007-2008: a multicentre European study.

    LENUS (Irish Health Repository)

    Wagner, K S

    2011-04-01

    Diphtheria is now rare in most European countries but, when cases do arise, the case fatality rate is high (5-10%). Because few countries continue to routinely screen for the causative organisms of diphtheria, the extent to which they are circulating amongst different European populations is largely unknown. During 2007-2008, ten European countries each screened between 968 and 8551 throat swabs from patients with upper respiratory tract infections. Six toxigenic strains of Corynebacterium diphtheriae were identified: two from symptomatic patients in Latvia (the country with the highest reported incidence of diphtheria in the European Union) and four from Lithuania (two cases, two carriers); the last reported case of diphtheria in Lithuania was in 2002. Carriage rates of non-toxigenic organisms ranged from 0 (Bulgaria, Finland, Greece, Ireland, Italy) to 4.0 per 1000 (95% CI 2.0-7.1) in Turkey. A total of 28 non-toxigenic strains were identified during the study (26 C. diphtheriae, one Corynebacterium ulcerans, one Corynebacterium pseudotuberculosis). The non-toxigenic C. ulcerans strain was isolated from the UK, the country with the highest reported incidence of cases due to C. ulcerans. Of the eleven ribotypes detected, Cluj was seen most frequently in the non-toxigenic isolates and, amongst toxigenic isolates, the major epidemic clone, Sankt-Petersburg, is still in circulation. Isolation of toxigenic C. diphtheriae and non-toxigenic C. diphtheriae and C. ulcerans in highly-vaccinated populations highlights the need to maintain microbiological surveillance, laboratory expertise and an awareness of these organisms amongst public health specialists, microbiologists and clinicians.

  17. REGULATION OF L-VALINE BIOSYNTHESIS IN CORYNEBACTERIUM GLUTAMICUM - ASPECTS OF PHYSIOLOGY AND BIOCHEMISTRY

    OpenAIRE

    Ilze Deniņa

    2010-01-01

    ANNOTATION This study investigates the optimum cellular physiology and intracellular biochemistry to achieve L-valine overproduction by Corynebacterium glutamicum recombinant strains. A transition in the bacterial growth rate to below maximum was found to be an optimum parameter of cellular physiology to increase L-valine synthesis rate by C. glutamicum during batch and fed-batch cultivations. The increase in acetohydroxyacid synthase activity was the determinant for the ...

  18. Engineering of Corynebacterium glutamicum for High-Yield l-Valine Production under Oxygen Deprivation Conditions

    OpenAIRE

    Hasegawa, Satoshi; Suda, Masako; Uematsu, Kimio; Natsuma, Yumi; Hiraga, Kazumi; Jojima, Toru; Inui, Masayuki; Yukawa, Hideaki

    2013-01-01

    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 gen...

  19. Feedback-Resistant Acetohydroxy Acid Synthase Increases Valine Production in Corynebacterium glutamicum

    OpenAIRE

    Elišáková, Veronika; Pátek, Miroslav; Holátko, Jiří; Nešvera, Jan; Leyval, Damien; Goergen, Jean-Louis; Delaunay, Stéphane

    2005-01-01

    Acetohydroxy acid synthase (AHAS), which catalyzes the key reactions in the biosynthesis pathways of branched-chain amino acids (valine, isoleucine, and leucine), is regulated by the end products of these pathways. The whole Corynebacterium glutamicum ilvBNC operon, coding for acetohydroxy acid synthase (ilvBN) and aceto hydroxy acid isomeroreductase (ilvC), was cloned in the newly constructed Escherichia coli-C. glutamicum shuttle vector pECKA (5.4 kb, Kmr). By using site-directed mutagenesi...

  20. Metabolic responses to pyruvate kinase deletion in lysine producing Corynebacterium glutamicum

    OpenAIRE

    Wittmann Christoph; Klopprogge Corinna; Becker Judith

    2008-01-01

    Abstract Background Pyruvate kinase is an important element in flux control of the intermediate metabolism. It catalyzes the irreversible conversion of phosphoenolpyruvate into pyruvate and is under allosteric control. In Corynebacterium glutamicum, this enzyme was regarded as promising target for improved production of lysine, one of the major amino acids in animal nutrition. In pyruvate kinase deficient strains the required equimolar ratio of the two lysine precursors oxaloacetate and pyruv...

  1. Selection and Characterization of a Lysine Yielding Mutant of Corynebacterium glutamicum - a Soil Isolate from Pakistan

    OpenAIRE

    Habib-ur-Rehman§٭, Abdul Hameed and Safia Ahmed

    2012-01-01

    L-lysine is the second limiting amino acid for poultry and supplemented in broiler feed for optimal performance. Lysine can be produced by inducing mutation in glutamate producing bacteria. The study was conducted to enhance lysine production from a local strain of Corynebacterium glutamicum. The bacterium was mutated by exposure to UV. Mutants resistant to s-2-aminoethyle L-cystein (AEC) and showing auxotrophy for L-homoserine were screened for lysine production qualitatively and quantitativ...

  2. Complete genome sequence of Corynebacterium pseudotuberculosis Cp31, isolated from an Egyptian buffalo

    DEFF Research Database (Denmark)

    Silva, Artur; Ramos, Rommel Thiago Jucá; Ribeiro Carneiro, Adriana;

    2012-01-01

    Corynebacterium pseudotuberculosis is of major veterinary importance because it affects many animal species, causing economically significant livestock diseases and losses. Therefore, the genomic sequencing of various lines of this organism, isolated from different hosts, will aid...... in the development of diagnostic methods and new prevention and treatment strategies and improve our knowledge of the biology of this microorganism. In this study, we present the genome of C. pseudotuberculosis Cp31, isolated from a buffalo in Egypt....

  3. In Silico Genome-Scale Reconstruction and Validation of the Corynebacterium glutamicum Metabolic Network

    DEFF Research Database (Denmark)

    Kjeldsen, Kjeld Raunkjær; Nielsen, J.

    2009-01-01

    A genome-scale metabolic model of the Gram-positive bacteria Corynebacterium glutamicum ATCC 13032 was constructed comprising 446 reactions and 411 metabolite, based on the annotated genome and available biochemical information. The network was analyzed using constraint based methods. The model...... and lactate. Comparable flux values between in silico model and experimental values were seen, although some differences in the phenotypic behavior between the model and the experimental data were observed,...

  4. Complete Genome Sequence of the Attenuated Corynebacterium pseudotuberculosis Strain T1.

    Science.gov (United States)

    Almeida, Sintia; Loureiro, Dan; Portela, Ricardo W; Mariano, Diego C B; Sousa, Thiago J; Pereira, Felipe L; Dorella, Fernanda A; Carvalho, Alex F; Moura-Costa, Lilia F; Leal, Carlos A G; Figueiredo, Henrique C; Meyer, Roberto; Azevedo, Vasco

    2016-01-01

    We present here the genome sequence of the attenuated Corynebacterium pseudotuberculosis strain T1. The sequencing was performed with an Ion Torrent Personal Genome Machine platform. The genome is a circular chromosome of 2,337,201 bp, with a G+C content of 52.85% and a total of 2,125 coding sequences (CDSs), 12 rRNAs, 49 tRNAs, and 24 pseudogenes. PMID:27609922

  5. Bioremediation of refinery wastewater using immobilised Burkholderia cepacia and Corynebacterium sp and their transconjugants

    OpenAIRE

    Abdullahi T. Ajao; Sabo E. Yakubu; Veronica J. Umoh; Joseph B. Ameh

    2013-01-01

    When oil spill occurs, it poses serious toxic hazards to all forms of life. Mixed culture of Burkholderia cepacia and Corynebacterium sp isolated from refinery sludge using selective enrichment technique was used for bioremediation of refinery wastewater in a laboratoryscale bioreactor. Physicochemical parameters of both raw and treated water were as determined and compared with Federal Environ - mental Protection Agency (FEPA-limit, Abuja, Nigeria) to asses the efficiency of the bioremediati...

  6. Recent advances in recombinant protein expression by Corynebacterium, Brevibacterium, and Streptomyces: from transcription and translation regulation to secretion pathway selection.

    Science.gov (United States)

    Liu, Long; Yang, Haiquan; Shin, Hyun-Dong; Li, Jianghua; Du, Guocheng; Chen, Jian

    2013-11-01

    Gram-positive bacteria are widely used to produce recombinant proteins, amino acids, organic acids, higher alcohols, and polymers. Many proteins have been expressed in Gram-positive hosts such as Corynebacterium, Brevibacterium, and Streptomyces. The favorable and advantageous characteristics (e.g., high secretion capacity and efficient production of metabolic products) of these species have increased the biotechnological applications of bacteria. However, owing to multiplicity from genes encoding the proteins and expression hosts, the expression of recombinant proteins is limited in Gram-positive bacteria. Because there is a very recent review about protein expression in Bacillus subtilis, here we summarize recent strategies for efficient expression of recombinant proteins in the other three typical Gram-positive bacteria (Corynebacterium, Brevibacterium, and Streptomyces) and discuss future prospects. We hope that this review will contribute to the development of recombinant protein expression in Corynebacterium, Brevibacterium, and Streptomyces. PMID:24068337

  7. Plasmid pGA1 from Corynebacterium glutamicum codes for a gene product that positively influences plasmid copy number.

    OpenAIRE

    Nesvera, J; Pátek, M; Hochmannová, J; Abrhámová, Z; Becvárová, V; Jelínkova, M; Vohradský, J

    1997-01-01

    The complete nucleotide sequence (4,826 bp) of the cryptic plasmid pGA1 from Corynebacterium glutamicum was determined. DNA sequence analysis revealed four putative coding regions (open reading frame A [ORFA], ORFA2, ORFB, and ORFC). ORFC was identified as a rep gene coding for an initiator of plasmid replication (Rep) according to the high level of homology of its deduced amino acid sequence with the Rep proteins of plasmids pSR1 (from C. glutamicum) and pNG2 (from Corynebacterium diphtheria...

  8. Linking Central Metabolism with Increased Pathway Flux: l-Valine Accumulation by Corynebacterium glutamicum

    OpenAIRE

    Radmacher, Eva; Vaitsikova, Adela; Burger, Udo; Krumbach, Karin; Sahm, Hermann; Eggeling, Lothar

    2002-01-01

    Mutants of Corynebacterium glutamicum were made and enzymatically characterized to clone ilvD and ilvE, which encode dihydroxy acid dehydratase and transaminase B, respectively. These genes of the branched-chain amino acid synthesis were overexpressed together with ilvBN (which encodes acetohydroxy acid synthase) and ilvC (which encodes isomeroreductase) in the wild type, which does not excrete l-valine, to result in an accumulation of this amino acid to a concentration of 42 mM. Since l-vali...

  9. l-Valine Production with Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum▿

    OpenAIRE

    Blombach, Bastian; Schreiner, Mark E.; Holátko, Jiří; Bartek, Tobias; Oldiges, Marco; Eikmanns, Bernhard J.

    2007-01-01

    Corynebacterium glutamicum was engineered for the production of l-valine from glucose by deletion of the aceE gene encoding the E1p enzyme of the pyruvate dehydrogenase complex and additional overexpression of the ilvBNCE genes encoding the l-valine biosynthetic enzymes acetohydroxyacid synthase, isomeroreductase, and transaminase B. In the absence of cellular growth, C. glutamicum ΔaceE showed a relatively high intracellular concentration of pyruvate (25.9 mM) and produced significant amount...

  10. Detection of Corynebacterium bovis infection in athymic nude mice from a research animal facility in Korea

    OpenAIRE

    Kim, Tae-Hyoun; Kim, Dong-Su; Han, Ju-Hee; Chang, Seo-Na; Kim, Kyung-Sul; Seok, Seung-Hyeok; Kim, Dong-Jae; Park, Jong-Hwan; Park, Jae-Hak

    2014-01-01

    Corynebacterium (C.) bovis infection in nude mice causes hyperkeratosis and weight loss and has been reported worldwide but not in Korea. In 2011, nude mice from an animal facility in Korea were found to have white flakes on their dorsal skin. Histopathological testing revealed that the mice had hyperkeratosis and Gram-positive bacteria were found in the skin. We identified isolated bacteria from the skin lesions as C. bovis using PCR and 16S rRNA sequencing. To the best of our knowledge, thi...

  11. Epidemiological survey of Corynebacterium equi infections on five Ontario horse farms.

    OpenAIRE

    Prescott, J. F.; Travers, M.; Yager-Johnson, J A

    1984-01-01

    Corynebacterium equi was cultured from manure or soil on five horse-breeding farms in Ontario at monthly intervals on three occasions during the summer of 1982. The organism was widespread. Contamination by C. equi of the loafing paddock and pasture areas was significantly greater in a farm established 30 years than in two established for four and six years and there was a significant correlation between the C. equi burden in stables, paddocks and pastures and the length of use of the five fa...

  12. Recurrent Breast Abscesses due to Corynebacterium kroppenstedtii, a Human Pathogen Uncommon in Caucasian Women

    Directory of Open Access Journals (Sweden)

    Anne Le Flèche-Matéos

    2012-01-01

    Full Text Available Background. Corynebacterium kroppenstedtii (Ck was first described in 1998 from human sputum. Contrary to what is observed in ethnic groups such as Maori, Ck is rarely isolated from breast abscesses and granulomatous mastitis in Caucasian women. Case Presentation. We herein report a case of recurrent breast abscesses in a 46-year-old Caucasian woman. Conclusion. In the case of recurrent breast abscesses, even in Caucasian women, the possible involvement of Ck should be investigated. The current lack of such investigations, probably due to the difficulty to detect Ck, may cause the underestimation of such an aetiology.

  13. Desulfurization of dibenzothiophene by a newly isolated Corynebacterium sp.ZD-1 in aqueous phase

    Institute of Scientific and Technical Information of China (English)

    WANG Miao-dong; LI Wei; WANG Da-hui; SHI Yao

    2004-01-01

    Sulfur emission through fuel combustion is a global problem because it is a major cause of acid rain. Crud oil contains many heterocyclic organic sulfur compounds, among which dibenzothiophene(DBT) and DBTs bearing alkyl substitutions usually are representative compounds. A strain was isolated from refinery sludge and identified as Corynebacterium ZD-1. The behavior of DBT degradation by ZD-1 in aqueous phase was investigated. Corynebacterium ZD-1 could metabolize DBT to 2-hydroxybiphenyl(2-HBP) as the dead-end metabolite through a sulfur-specific pathway. In shake flask culture, ZD-1 had its maximal desulfurization activity in the late exponential growth phase and the specific production rate of 2-HBP was about 0.14(mmol·kg dry cell-1·min-1, mmol·KDC-1·min-1). Active resting cells for desulfurization should be prepared only in this period. 2-HBP inhibited the growth of strain ZD-1, the production of DBT degradation enzymes, and the activity of enzymes. Sulfate inhibited the production of dibenzothiophene(DBT) degradation enzymes but had no effect on the enzymes' activity. The production rates of 2-HBP at lower cell densities were higher and the maximum amount conversion of DBT to 2-HBP(0.067 mmol/L) after 8 h was gained at 9.2(g dry cell/L) rather higher cell density. The results indicated that this newly isolated strain could be a promising biocatalyst for DBT desulfurization.

  14. Corynebacterium uropygiale sp. nov., isolated from the preen gland of Turkeys (Meleagris gallopavo).

    Science.gov (United States)

    Braun, Markus Santhosh; Zimmermann, Stefan; Danner, Maria; Rashid, Harun-or; Wink, Michael

    2016-03-01

    A novel species of fastidious, lipophilic, club-shaped, Gram-positive bacteria was recovered from the preen glands of healthy Turkeys (Meleagris gallopavo) from two different locations. Phylogenetic analysis of the 16S rRNA gene showed highest similarity to Corynebacterium spheniscorum DSM 44757(T) (96.8%) with a 3.2kb stretch of rpoB sharing 82.4% sequence similarity to the same species. DNA fingerprinting by ERIC-PCR and polar lipid profiles clearly differentiated the Turkey isolates from the most closely related Corynebacteria, as did MALDI-TOF MS analysis. Chemotaxonomic tests revealed the presence of corynemycolic acids with C16:0, C18:0, C18:1ω9c and tuberculostearic acid as the major cellular fatty acids. The G+C content of the type strain was 60.7 mol%. The species was susceptible to ampicillin, kanamycin A, streptomycin, amikacin, polymyxin B and vancomycin. From our results, it becomes evident that the isolated organisms represent a new species, for which the name Corynebacterium uropygiale sp. nov. is proposed. The type strain is Iso10(T) (=DSM 46817(T)=LMG 28616(T)).

  15. Systems pathway engineering of Corynebacterium crenatum for improved L-arginine production.

    Science.gov (United States)

    Man, Zaiwei; Xu, Meijuan; Rao, Zhiming; Guo, Jing; Yang, Taowei; Zhang, Xian; Xu, Zhenghong

    2016-01-01

    L-arginine is an important amino acid in food and pharmaceutical industries. Until now, the main production method of L-arginine in China is the highly polluting keratin acid hydrolysis. The industrial level L-arginine production by microbial fermentation has become an important task. In previous work, we obtained a new L-arginine producing Corynebacterium crenatum (subspecies of Corynebacterium glutamicum) through screening and mutation breeding. In this work, we performed systems pathway engineering of C. crenatum for improved L-arginine production, involving amplification of L-arginine biosynthetic pathway flux by removal of feedback inhibition and overexpression of arginine operon; optimization of NADPH supply by modulation of metabolic flux distribution between glycolysis and pentose phosphate pathway; increasing glucose consumption by strengthening the preexisting glucose transporter and exploitation of new glucose uptake system; channeling excess carbon flux from glycolysis into tricarboxylic acid cycle to alleviate the glucose overflow metabolism; redistribution of carbon flux at α-ketoglutarate metabolic node to channel more flux into L-arginine biosynthetic pathway; minimization of carbon and cofactor loss by attenuation of byproducts formation. The final strain could produce 87.3 g L(-1) L-arginine with yield up to 0.431 g L-arginine g(-1) glucose in fed-batch fermentation. PMID:27338253

  16. Systems pathway engineering of Corynebacterium crenatum for improved L-arginine production

    Science.gov (United States)

    Man, Zaiwei; Xu, Meijuan; Rao, Zhiming; Guo, Jing; Yang, Taowei; Zhang, Xian; Xu, Zhenghong

    2016-01-01

    L-arginine is an important amino acid in food and pharmaceutical industries. Until now, the main production method of L-arginine in China is the highly polluting keratin acid hydrolysis. The industrial level L-arginine production by microbial fermentation has become an important task. In previous work, we obtained a new L-arginine producing Corynebacterium crenatum (subspecies of Corynebacterium glutamicum) through screening and mutation breeding. In this work, we performed systems pathway engineering of C. crenatum for improved L-arginine production, involving amplification of L-arginine biosynthetic pathway flux by removal of feedback inhibition and overexpression of arginine operon; optimization of NADPH supply by modulation of metabolic flux distribution between glycolysis and pentose phosphate pathway; increasing glucose consumption by strengthening the preexisting glucose transporter and exploitation of new glucose uptake system; channeling excess carbon flux from glycolysis into tricarboxylic acid cycle to alleviate the glucose overflow metabolism; redistribution of carbon flux at α-ketoglutarate metabolic node to channel more flux into L-arginine biosynthetic pathway; minimization of carbon and cofactor loss by attenuation of byproducts formation. The final strain could produce 87.3 g L−1 L-arginine with yield up to 0.431 g L-arginine g−1 glucose in fed-batch fermentation. PMID:27338253

  17. Growth characteristics of Brevibacterium, Corynebacterium, Microbacterium, and Staphylococcus spp. isolated from surface-ripened cheese.

    Science.gov (United States)

    Mounier, Jérôme; Rea, Mary C; O'Connor, Paula M; Fitzgerald, Gerald F; Cogan, Timothy M

    2007-12-01

    The growth characteristics of five bacteria, Brevibacterium aurantiacum 1-16-58, Corynebacterium casei DPC 5298(T), Corynebacterium variabile DPC 5310, Microbacterium gubbeenense DPC 5286(T), and Staphylococcus saprophyticus 4E61, all of which were isolated from the surface of smear cheese, were studied in complex and chemically defined media. All of the coryneforms, except M. gubbeenense, grew in 12% salt, while B. aurantiacum and S. saprophyticus grew in 15% salt. All five bacteria assimilated lactate in a semisynthetic medium, and none of the coryneform bacteria assimilated lactose. Glucose assimilation was poor, except by S. saprophyticus and C. casei. Five to seven amino acids were assimilated by the coryneforms and 12 by S. saprophyticus. Glutamate, phenylalanine, and proline were utilized by all five bacteria, whereas utilization of serine, threonine, aspartate, histidine, alanine, arginine, leucine, isoleucine, and glycine depended on the organism. Growth of C. casei restarted after addition of glutamate, proline, serine, and lactate at the end of the exponential phase, indicating that these amino acids and lactate can be used as energy sources. Pantothenic acid was essential for the growth of C. casei and M. gubbeenense. Omission of biotin reduced the growth of B. aurantiacum, C. casei, and M. gubbeenense. All of the bacteria contained lactate dehydrogenase activity (with both pyruvate and lactate as substrates) and glutamate pyruvate transaminase activity but not urease activity.

  18. Toxigenic Corynebacterium ulcerans isolated from a hunting dog and its diphtheria toxin antibody titer.

    Science.gov (United States)

    Katsukawa, Chihiro; Komiya, Takako; Umeda, Kaoru; Goto, Minami; Yanai, Tokuma; Takahashi, Motohide; Yamamoto, Akihiko; Iwaki, Masaaki

    2016-03-01

    Toxigenic Corynebacterium ulcerans is a zoonotic pathogen that produces diphtheria toxin and causes a diphtheria-like illness in humans. The organism is known to infect and circulate among dogs, which can then transmit it to humans. Furthermore, previous studies have found that C. ulcerans is carried by wild animals, including game animals. In the present study, we tested hunting and companion dogs for the presence of toxigenic C. ulcerans and succeeded in isolating the bacterium from a hunting dog. Moreover, several hunting dogs had serum diphtheria antitoxin titers that were higher than the titers required for protection in humans, suggesting a history of exposure to toxigenic Corynebacterium strains. Notably, ribotyping, pulsed-field gel electrophoresis and tox gene sequencing demonstrated that the isolate from the hunting dog clustered with previously characterized C. ulcerans strains isolated from wild animals, as opposed to groups of isolates from humans and companion dogs. Interestingly, the wild animal cluster also contains an isolate from an outdoor breeding dog, which could have formed a bridge between isolates from wild animals and those from companion dogs. The results presented herein provide insight into the mechanism by which the zoonotic pathogen C. ulcerans circulates among wild animals, hunting and companion dogs, and humans. PMID:26853714

  19. Draft Genome Sequence of Toxigenic Corynebacterium ulcerans Strain 04-7514, Isolated from a Dog in France

    Science.gov (United States)

    Viana, Marcus V. C.; Benevides, Leandro J.; Mariano, Diego C. B.; Veras, Adooney A. O.; Sá, Pablo H. C.; Rocha, Flávia S.; Vilas Boas, Priscilla C. B.; Soares, Siomar C.; Barbosa, Maria S.; Guiso, Nicole; Badell, Edgar; Carneiro, Adriana R.; Azevedo, Vasco; Ramos, Rommel T. J.

    2016-01-01

    Here, we present the draft genome of toxigenic Corynebacterium ulcerans strain 04-7514. The draft genome has 2,497,845 bp, 2,059 coding sequences, 12 rRNA genes, 46 tRNA genes, 150 pseudogenes, 1 clustered regularly interspaced short palindromic repeat (CRISPR) array, and a G+C content of 53.50%. PMID:27034487

  20. Genome sequence of Corynebacterium pseudotuberculosis biovar equi strain 258 and prediction of antigenic targets to improve biotechnological vaccine production

    DEFF Research Database (Denmark)

    Soares, Siomar C; Trost, Eva; Ramos, Rommel T J;

    2013-01-01

    Corynebacterium pseudotuberculosis is the causative agent of several veterinary diseases in a broad range of economically important hosts, which can vary from caseous lymphadenitis in sheep and goats (biovar ovis) to ulcerative lymphangitis in cattle and horses (biovar equi). Existing vaccines ag...

  1. Genome Sequence of Corynebacterium glutamicum ATCC 14067, Which Provides Insight into Amino Acid Biosynthesis in Coryneform Bacteria

    OpenAIRE

    Lv, Yangyong; Liao, Juanjun; Wu, Zhanhong; Han, Shuangyan; Lin, Ying; Zheng, Suiping

    2012-01-01

    We report the genome sequence of Corynebacterium glutamicum ATCC 14067 (once named Brevibacterium flavum), which is useful for taxonomy research and further molecular breeding in amino acid production. Preliminary comparison with those of the reported coryneform strains revealed some notable differences that might be related to the difficulties in molecular manipulation.

  2. Draft Genome Sequence of Corynebacterium variabile Mu292, Isolated from Munster, a French Smear-Ripened Cheese

    Science.gov (United States)

    Sarthou, Anne-Sophie; Loux, Valentin; Vidal, Marie; Bonnarme, Pascal; Irlinger, Françoise

    2016-01-01

    Here, we report the draft genome sequence of Corynebacterium variabile Mu292, which was originally isolated from the surface of Munster, a French smear-ripened cheese. This genome investigation will improve our knowledge on the molecular determinants potentially involved in the adaptation of this strain during the Munster-type cheese manufacturing process. PMID:27445372

  3. Osmolality, temperature, and membrane lipid composition modulate the activity of betaine transporter BetP in Corynebacterium glutamicum

    DEFF Research Database (Denmark)

    Ozcan, Nuran; Ejsing, Christer S.; Shevchenko, Andrej;

    2007-01-01

    The gram-positive soil bacterium Corynebacterium glutamicum, a major amino acid-producing microorganism in biotechnology, is equipped with several osmoregulated uptake systems for compatible solutes, which is relevant for the physiological response to osmotic stress. The most significant carrier,...... dynamics by local anesthetics and the lack of a possible influence of internally accumulated betaine on BetP activity....

  4. Complete Genome Sequence of Corynebacterium camporealensis DSM 44610, Isolated from the Milk of a Manchega Sheep with Subclinical Mastitis.

    Science.gov (United States)

    Rückert, Christian; Albersmeier, Andreas; Winkler, Anika; Tauch, Andreas

    2015-01-01

    Corynebacterium camporealensis has been isolated in pure culture from milk samples of dairy sheep affected by subclinical mastitis. The complete genome sequence of the type strain DSM 44610, recovered from milk of a Manchega sheep, comprises 2,451,810 bp with a mean G+C content of 59.41% and 2,249 protein-coding genes. PMID:26021938

  5. Complete Genome Sequence of Corynebacterium camporealensis DSM 44610, Isolated from the Milk of a Manchega Sheep with Subclinical Mastitis

    OpenAIRE

    Rückert, Christian; Albersmeier, Andreas; Winkler, Anika; Tauch, Andreas

    2015-01-01

    Corynebacterium camporealensis has been isolated in pure culture from milk samples of dairy sheep affected by subclinical mastitis. The complete genome sequence of the type strain DSM 44610, recovered from milk of a Manchega sheep, comprises 2,451,810 bp with a mean G+C content of 59.41% and 2,249 protein-coding genes.

  6. In Vitro Susceptibility of Equine-Obtained Isolates of Corynebacterium pseudotuberculosis to Gallium Maltolate and 20 Other Antimicrobial Agents

    Science.gov (United States)

    Batista, M.; Lawhon, S. D.; Zhang, S.; Kuskie, K. R.; Swinford, A. K.; Bernstein, L. R.; Cohen, N. D.

    2014-01-01

    This study's objective was to determine the in vitro antimicrobial activities of gallium maltolate (GaM) and 20 other antimicrobial agents against clinical equine isolates of Corynebacterium pseudotuberculosis. The growth of cultured isolates was not inhibited by any concentration of GaM. MIC data revealed susceptibility to commonly used antimicrobials. PMID:24829243

  7. Teste de pele em caprinos vacinados e infectados com Corynebacterium pseudotuberculosis Skin test of goats vaccinated and infected with Corynebacterium pseudotuberculosis

    Directory of Open Access Journals (Sweden)

    Francisco Selmo Fernandes Alves

    1999-07-01

    Full Text Available Dez caprinos foram vacinados com toxóide a 3%, outros dez com uma bacterina e mais dois grupos-controle de cinco animais cada, submetidos à inoculação de infusão de cérebro e coração e solução salina, respectivamente. Todos os animais foram examinados e avaliados com um teste de pele. Tanto o toxóide quanto a bacterina foram produzidos a partir de amostra de Corynebacterium pseudotuberculosis. Todos os caprinos foram desafiados com C. pseudotuberculosis, trinta dias após as vacinações. Nenhuma das vacinas induziu reação de hipersensibilidade na pele dos caprinos antes do desafio. Após o desafio, todos os animais desenvolveram reações mensuráveis na primeira, quinta e décima semana em resposta ao teste de pele. Os diâmetros da reação dérmica aumentaram do décimo dia à quinta semana após o desafio. As medidas alcançaram tamanho maior na décima semana. O resultado deste estudo indica que antígeno específico do C. pseudotuberculosis pode ser utilizado em caprinos no diagnóstico da linfadenite caseosa como teste de pele ou como instrumento experimental para monitorar o desenvolvimento da doença.Ten goats were vaccinated with a 3% toxoid, ten vaccinated with a bacterin and two control groups (five animals each inoculated with brain heart infusion and saline solution, respectively. All animals were skin tested with a crude antigen of formalin-killed Corynebacterium pseudotuberculosis bacterial cells. All goats were challenged with a virulent C. pseudotuberculosis thirty days after vaccination. Neither the vaccinated nor control goats responded to the skin test prior to infection. After the challenge, dermal reactions were demonstrated in all animals at one week, five and ten weeks. The diameters increased from the first week, five and ten weeks. The reactions were more proeminent at ten weeks. The results of this study indicate that skin testing with a specific bacterial antigen of C. pseudotuberculosis may be useful

  8. Toxigenic Corynebacterium ulcerans isolated from a wild bird (ural owl) and its feed (shrew-moles): comparison of molecular types with human isolates

    OpenAIRE

    Katsukawa, Chihiro; Umeda, Kaoru; Inamori, Ikuko; Kosono, Yuka; Tanigawa, Tomokazu; Komiya, Takako; Iwaki, Masaaki; Yamamoto, Akihiko; Nakatsu, Susumu

    2016-01-01

    Background Corynebacterium ulcerans is a pathogen causing diphtheria-like illness to humans. In contrast to diphtheria by Corynebacterium diphtheriae circulating mostly among humans, C. ulcerans infection is zoonotic. The present study aimed to clarify how a zoonotic pathogen C. ulcerans circulates among wild birds and animals. Results By screening 380 birds, a single strain of toxigenic C. ulcerans was isolated from a carnivorous bird, ural owl (Strix uralensis). The bacterium was also isola...

  9. Global Transcriptomic Analysis of the Response of Corynebacterium glutamicum to Vanillin

    Science.gov (United States)

    Chen, Can; Pan, Junfeng; Yang, Xiaobing; Guo, Chenghao; Ding, Wei; Si, Meiru; Zhang, Yi; Shen, Xihui; Wang, Yao

    2016-01-01

    Lignocellulosic biomass is an abundant and renewable resource for biofuels and bio-based chemicals. Vanillin is one of the major phenolic inhibitors in biomass production using lignocellulose. To assess the response of Corynebacterium glutamicum to vanillin stress, we performed a global transcriptional response analysis. The transcriptional data showed that the vanillin stress not only affected the genes involved in degradation of vanillin, but also differentially regulated several genes related to the stress response, ribosome/translation, protein secretion, and the cell envelope. Moreover, deletion of the sigH or msrA gene in C. glutamicum resulted in a decrease in cell viability under vanillin stress. These insights will promote further engineering of model industrial strains, with enhanced tolerance or degradation ability to vanillin to enable suitable production of biofuels and bio-based chemicals from lignocellulosic biomass. PMID:27760214

  10. Molecular cloning and expression of Corynebacterium glutamicum genes for amino acid synthesis in Escherichia coli cells

    International Nuclear Information System (INIS)

    Molecular cloning of Corynebacterium glutamicum genes for threonine and lysine synthesis has been done in Escherichia coli cells. The clonal library of EcoRI fragments of chromosomal DNA of C. glutamicum was constructed on the plasmid vector λpSL5. The genes for threonine and lysine synthesis were identified by complementation of E. coli mutations in thrB and lysA genes, respectively. Recombinant plasmids, isolated from independent ThrB+ clone have a common 4.1-kb long EcoRI DNA fragment. Hybrid plasmids isolated from LysA+ transductants of E. coli have common 2.2 and 3.3 kb long EcoRI fragments of C. glutamicum DNA. The hybrid plasmids consistently transduced the markers thrB+ and lysA+. The Southern hybridization analysis showed that the cloned DNA fragments hybridized with the fragments of identical length in C. glutamicum chromosomes

  11. BIOCHEMICAL AND PHYLOGENETIC STUDIES OF CreD OF Corynebacterium glutamicum

    Directory of Open Access Journals (Sweden)

    Muhammad Tausif Chaudhry

    2015-06-01

    Full Text Available CreD characterized as Mg2+-dependent phosphohydrolase with conserved HD domain was involved in 4-cresol metabolism in Corynebacterium glutamicum. Native molecular mass of 54 kDa suggested that the biological unit is a dimer. No deoxynucleotide triphosphate triphosphohydrolase (dNTPase activity was detected for CreD. The apparent Km and Vmax values for 4-nitrophenyl phosphate were 0.35 mM and 16.23 M min-1 mg-1, respectively, while calculated values for kcat and kcat/Km were 0.4 s-1 and 1.14103 M-1 s-1, respectively. Among thiol group inhibitors, iodoacetic acid significantly inhibited phosphohydrolase activity. Sequence identity and phylogenetic analysis suggested universal existence of CreD homologues. Involvement of HD-domain hydrolase in aromatic degradation has not been reported before.

  12. Tips and tricks for the assembly of a Corynebacterium pseudotuberculosis genome using a semiconductor sequencer

    DEFF Research Database (Denmark)

    Ramos, Rommel Thiago Jucá; Carneiro, Adriana Ribeiro; Soares, Siomar de Castro;

    2013-01-01

    New sequencing platforms have enabled rapid decoding of complete prokaryotic genomes at relatively low cost. The Ion Torrent platform is an example of these technologies, characterized by lower coverage, generating challenges for the genome assembly. One particular problem is the lack of genomes ...... is not a traditional biological model such as Escherichia coli.......New sequencing platforms have enabled rapid decoding of complete prokaryotic genomes at relatively low cost. The Ion Torrent platform is an example of these technologies, characterized by lower coverage, generating challenges for the genome assembly. One particular problem is the lack of genomes...... that enable reference-based assembly, such as the one used in the present study, Corynebacterium pseudotuberculosis biovar equi, which causes high economic losses in the US equine industry. The quality treatment strategy incorporated into the assembly pipeline enabled a 16-fold greater use of the sequencing...

  13. Effects of non-specific immunopotentiators in experimental Schistosoma mansoni infection. II. Corynebacterium parvum.

    Science.gov (United States)

    Teixeira, K M; Coutinho, E M; Abath, F G; Montenegro, S M

    1996-01-01

    The effects of Corynebacterium parvum on host protection, tissue reaction and "in vivo" chemotaxis in Schistosoma mansoni infected mice were studied. The C. parvum was given intraperitoneally using a dose of 0.7 mg, twice a week (for 4 weeks), thirty days before (prophylactic treatment) or after infection (curative treatment). The host protection was evaluated through the recovery of adult worms by liver perfusion and was lower in the prophylactic group as compared to the control group (p = 0.018), resulting in 44% protection. The "in vivo" leukocyte response in both prophylactic and curative groups was higher as compared to the infected/non treated group (p = 0.009 and p = 0.003, respectively). Tissue reactions were described in the experimental and control groups, but there were not remarkable differences among them. The possible biological implications and relevance of the findings for the defensive response of the host and control of schistosomiasis are discussed. PMID:9293078

  14. Influence of Corynebacterium parvum on the phagocytosis of 198Au colloids in rats

    International Nuclear Information System (INIS)

    The kinetics of the phagocytosis of gelatin-protected 198Au colloids in Wistar rats treated with Corynebacterium Parvum (CBP), was studied in order to explain its mechanism of immunomodulation. A previously developed extracorporeal blood circulation technique was used. The changes in the rate of phagocytosis, v, after the administration of CBP, for a dose of the 198Au colloid smaller or higher than the substratum constant, were studied. In the first case, no significant changes of v were observed; in the second case, significant increases of v were determined, which reached a maximum 6 days after the CBP administration. The kinetic analysis of the obtained data indicates that the action of CBP is exerted on the stage of the entrance of the colloidal particle into the reticuloendothelial cell. (author)

  15. Bioconversion of sugar cane molasses into glutamic acid by gamma irradiated corynebacterium glutamicum

    International Nuclear Information System (INIS)

    Corynebacterium glutamicum (ATCC 13058) was used for glutamic acid production from sugar cane molasses which contain sufficient. The addition of 5 units ml4 of penicillin G was superior in glutamic acid production (11.5 g L 4). Tweens and their saturated fatty acids were effective on the accumulation of glutamic acid in the culture medium and the maximum yield (16.6 g L4) was the addition of 5 mg ml4 Tween 40. Gamma irradiation prior to Tween-40 treatment of bacterial cells resulted in an obvious increase in glutamic acid production and it was maximum (23.72 g L4) at 0.1 k Gy exposure dose of inocula. 5 tabs

  16. Toxigenic Corynebacterium ulcerans isolated from a free-roaming red fox (Vulpes vulpes).

    Science.gov (United States)

    Sting, Reinhard; Ketterer-Pintur, Sandra; Contzen, Matthias; Mauder, Norman; Süss-Dombrowski, Christine

    2015-01-01

    Corynebacterium (C.) ulcerans could be isolated from the spleen of a red fox (Vulpes vulpes) that had been found dead in the state of Baden-Württemberg, Germany. Pathohistological examination suggested that the fox had died of distemper, as was confirmed by PCR. The isolate was identified biochemically, by MALDI-TOF MS, FT-IR and by partial 16S rRNA, rpoB and tox gene sequencing. Using the Elek test the C. ulcerans isolate demonstrated diphtheria toxin production. FT-IR and sequencing data obtained from the C. ulcerans isolate from the red fox showed higher similarity to isolates from humans than to those from wild game. PMID:26054226

  17. Surface modification of polyacrylonitrile fibre by nitrile hydratase from Corynebacterium nitrilophilus.

    Science.gov (United States)

    Chen, Sheng; Gao, Huihui; Chen, Jian; Wu, Jing

    2014-11-01

    Previously, nitrile hydratase (NHase) from Corynebacterium nitrilophilus was obtained and showed potential in polyacrylonitrile (PAN) fibre modification. In the present study, the modification conditions of C. nitrilophilus NHase on PAN were investigated. In the optimal conditions, the wettability and dyeability (anionic and reactive dyes) of PAN treated by C. nitrilophilus NHase reached a similar level of those treated by alkali. In addition, the chemical composition and microscopically observable were changed in the PAN surface after NHase treatment. Meanwhile, it revealed that cutinase combined with NHase facilitates the PAN hydrolysis slightly because of the ester existed in PAN as co-monomer was hydrolyzed. All these results demonstrated that C. nitrilophilus NHase can modify PAN efficiently without textile structure damage, and this study provides a foundation for the further application of C. nitrilophilus NHase in PAN modification industry. PMID:25163886

  18. Corynebacterium glutamicum superoxide dismutase is a manganese-strict non-cambialistic enzyme in vitro.

    Science.gov (United States)

    El Shafey, H M; Ghanem, S; Merkamm, M; Guyonvarch, A

    2008-01-01

    Superoxide dismutase (SOD) of Corynebacterium glutamicum was purified and characterized. The enzyme had a native molecular weight of about 80kDa, whereas a monomer with molecular weight of 24kDa was found on SDS-PAGE suggesting it to be homotetramer. The native SOD activity stained gel revealed a unique cytosolic enzyme. Supplementing growth media with manganese increased the specific activity significantly, while adding iron did not result in significant difference. No growth perturbation was observed with the supplemented media. In vitro metal removal and replacement studies revealed conservation of about 85% of the specific activity by substitution with manganese, while substitution with copper, iron, nickel or zinc did not restore any significant specific activity. Manganese was identified by atomic absorption spectrometer, while no signals corresponding to fixing other metallic elements were detected. Thus, C. glutamicum SOD could be considered a strict (non-cambialistic) manganese superoxide dismutase (MnSOD). PMID:16809027

  19. Pathogenicity and genetic variation of 3 strains of Corynebacterium bovis in immunodeficient mice.

    Science.gov (United States)

    Dole, Vandana S; Henderson, Kenneth S; Fister, Richard D; Pietrowski, Michael T; Maldonado, Geomaris; Clifford, Charles B

    2013-07-01

    Corynebacterium bovis has been associated with hyperkeratotic dermatitis and acanthosis in mice. We studied 3 different strains of C. bovis: one previously described to cause hyperkeratotic dermatitis (HAC), one that infected athymic nude mice without leading to the classic clinical signs, and one of bovine origin (ATCC 7715). The 3 strains showed a few biochemical and genetic differences. Immunodeficient nude mice were housed in 3 independent isolators and inoculated with pure cultures of the 3 strains. We studied the transmission of these C. bovis studies to isolator-bedding and contact sentinels housed for 5 to 12 wk in filter-top or wire-top cages in the respective isolators. Using a 16S rRNA-based qPCR assay, we did not find consistent differences in growth and transmission among the 3 C. bovis strains, and neither the incidence nor severity of hyperkeratosis or acanthosis differed between strains. Housing in filter-top compared with wire-top cages did not alter the morbidity associated with any of the strains. Our findings confirmed the variability in the gross and histologic changes associated with C. bovis infection of mice. Although bacteriology was a sensitive method for the detection of Corynebacterium spp., standard algorithms occasionally misidentified C. bovis and several related species. Our study demonstrates that PCR of skin swabs or feces is a sensitive and specific method for the detection of C. bovis infection in mice. An rpoB-based screen of samples from North American vivaria revealed that HAC is the predominant C. bovis strain in laboratory mice. PMID:23849444

  20. Evidence for reductive genome evolution and lateral acquisition of virulence functions in two Corynebacterium pseudotuberculosis strains.

    Directory of Open Access Journals (Sweden)

    Jerônimo C Ruiz

    Full Text Available BACKGROUND: Corynebacterium pseudotuberculosis, a gram-positive, facultative intracellular pathogen, is the etiologic agent of the disease known as caseous lymphadenitis (CL. CL mainly affects small ruminants, such as goats and sheep; it also causes infections in humans, though rarely. This species is distributed worldwide, but it has the most serious economic impact in Oceania, Africa and South America. Although C. pseudotuberculosis causes major health and productivity problems for livestock, little is known about the molecular basis of its pathogenicity. METHODOLOGY AND FINDINGS: We characterized two C. pseudotuberculosis genomes (Cp1002, isolated from goats; and CpC231, isolated from sheep. Analysis of the predicted genomes showed high similarity in genomic architecture, gene content and genetic order. When C. pseudotuberculosis was compared with other Corynebacterium species, it became evident that this pathogenic species has lost numerous genes, resulting in one of the smallest genomes in the genus. Other differences that could be part of the adaptation to pathogenicity include a lower GC content, of about 52%, and a reduced gene repertoire. The C. pseudotuberculosis genome also includes seven putative pathogenicity islands, which contain several classical virulence factors, including genes for fimbrial subunits, adhesion factors, iron uptake and secreted toxins. Additionally, all of the virulence factors in the islands have characteristics that indicate horizontal transfer. CONCLUSIONS: These particular genome characteristics of C. pseudotuberculosis, as well as its acquired virulence factors in pathogenicity islands, provide evidence of its lifestyle and of the pathogenicity pathways used by this pathogen in the infection process. All genomes cited in this study are available in the NCBI Genbank database (http://www.ncbi.nlm.nih.gov/genbank/ under accession numbers CP001809 and CP001829.

  1. Corynebacterium striatum infecting a malignant cutaneous lesion: the emergence of an opportunistic pathogen Corynebacterium striatum infectando lesão cutânea maligna: a emergência de um patógeno oportunista

    Directory of Open Access Journals (Sweden)

    Silvana Vargas Superti

    2009-04-01

    Full Text Available We described a case of a 27-year old male patient with skin and soft tissue infection of a neoplastic lesion caused by Corynebacterium striatum, an organism which has been rarely described as a human pathogen. Identification was confirmed by DNA sequencing. Successful treatment with penicillin was achieved. The role of the C. striatum as an emerging opportunistic pathogen is discussed.Descrevemos infecção de lesão neoplásica em paciente masculino de 27 anos, envolvendo pele e partes moles, causada por Corynebacterium striatum, um microrganismo raramente descrito como patógeno humano. A identificação foi confirmada por seqüenciamento de DNA. O paciente foi tratado com penicilina, com sucesso. O papel do C. striatum como patógeno oportunista é discutido.

  2. Effects of non-specific immunopotentiators in experimental Schistosoma mansoni infection: II. Corynebacterium parvum Efeitos de imunopotenciadores não específicos na infecção experimental pelo Schistosoma mansoni: II. Corynebacterium parvum

    OpenAIRE

    Kirte M Teixeira; Eridan M. Coutinho; Frederico G.C. Abath; Montenegro, Silvia M. L.

    1996-01-01

    The effects of Corynebacterium parvum on host protection, tissue reaction and "in vivo" chemotaxis in Schistosoma mansoni infected mice were studied. The C. parvum was given intraperitoneally using a dose of 0.7 mg, twice a week (for 4 weeks), thirty days before (prophylactic treatment) or after infection (curative treatment). The host protection was evaluated through the recovery of adult worms by liver perfusion and was lower in the prophylactic group as compared to the control group (p = 0...

  3. A single method to stain Malassezia furfur and Corynebacterium minutissimum in scales Um método simples para corar Malassezia furfur e Corynebacterium minutissimum nas escamas

    Directory of Open Access Journals (Sweden)

    Antar Padilha-Gonçalves

    1996-08-01

    Full Text Available A single and practical method to slain Malassezia furfur and Corynebacterium minutissimum in lesions' scales is described. The scales are collected by pressing small pieces of scotch tape (about 4 cm lenght and 2 cm width onto the lesions and following withdrawl the furfuraceous scales will remain on the glue side. These pieces are then immersed for some minutes in lactophenol-cotton blue stain. Following absorption of the stain the scales are washed in current water to remove the excess of blue stain, dried with filter paper, dehydrated via passage in two bottles containing absolute alcohol and then placed in xylene in a centrifugation tube. The xylene dissolves the scotch tape glue and the scales fall free in the tube. After centrifugation and decantation the scales concentrated on the bottom of the tube are collected with a platinum-loop, placed in Canada balsam on a microscopy slide and closed with a cover slip. The preparations are then ready to be submitted to microscopic examination. Other stains may also be used instead of lactophenol-cotton blue. This method is simple, easily performed, and offers good conditions to study these fungi as well as being useful for the diagnosis of the diseases that they cause.É descrito um método simples e prático para corar Malassezia furfur e Corynebacterium minutissimum nas escamas das lesões. O material é colhido com o auxílio de fita durex que será usada na maior parte das etapas do método para ajudar a fácil execução do processo de coloração. Para colher as escamas, pequenos pedaços de fita durex com cerca de 4 cm de comprimento por 2 cm de largura são colocados e pressionados sobre as lesões, e quando retirados trazem aderidas as escamas furfuráceas na face com goma. Esses pedaços de fita durex são imersos por alguns minutos no corante lactofenol-azul cotton e logo que as escamas estiverem coradas em azul são lavadas em água corrente para remover o excesso de corante azul, secos

  4. Molecular cloning, DNA sequence analysis, and characterization of the Corynebacterium diphtheriae dtxR homolog from Brevibacterium lactofermentum.

    OpenAIRE

    Oguiza, J A; Tao, X; Marcos, A T; Martín, J F; Murphy, J R

    1995-01-01

    A homolog of the Corynebacterium diphtheriae dtxR gene was isolated from Brevibacterium lactofermentum. The product of the B. lactofermentum dtxR gene was immunoreactive with polyclonal anti-DtxR antibodies and functioned as an iron-activated repressor capable of regulating the expression of beta-galactosidase from a diphtheria tox promoter/operator transcriptional fusion in recombinant Escherichia coli. The extents of induction by increasing concentrations of the chelator 2,2'-dipyridyl were...

  5. Malign Plevral Efüzyonların Önlenmesinde Corynebacterium Parvum’un Rolü

    OpenAIRE

    BEDİRHAN, M.A.; ÇAĞLAR, T.; Canbaz, S; MEHMET, R.

    2010-01-01

    THE ROLE OF CORYNEBACTERİUM PARVUM PLEURODESIS IN MALIGN PLEURAL EFFUSIONS §/ Inactivated form of Corynebacterium parvum (CBP), which is a gram positive anaerobic bacteria and besides its sclerotic effect in the pleuro has cytotoxics, antitumour effect, is used in symptomatic treatment of malign pleural effusions. Twentyseven CBP pleurodesis, in 20 patients, were reviewed retrospectively. A second pleurodesis was necessary in three cases. In one case response was partial and no response in...

  6. Native Valve Endocarditis due to Corynebacterium striatum confirmed by 16S Ribosomal RNA Sequencing: A Case Report and Literature Review

    Science.gov (United States)

    2016-01-01

    Corynebacterium species are non-fermentous Gram-positive bacilli that are normal flora of human skin and mucous membranes and are commonly isolated in clinical specimens. Non-diphtheriae Corynebacterium are regarded as contaminants when found in blood culture. Currently, Corynebacterium striatum is considered one of the emerging nosocomial agents implicated in endocarditis and serious infections. We report a case of native-valve infective endocarditis caused by C. striatum, which was misidentified by automated identification system but identified accurately by 16S ribosomal RNA sequencing, in a 55-year-old male patient. The patient had two mobile vegetations on his mitral valve, both of which had high embolic risk. Through surgical valve replacement and an antibiotic regimen, the patient recovered completely. In unusual clinical scenarios, C. striatum should not be simply dismissed as a contaminant when isolated from clinical specimens. The possibility of C. striatum infection should be considered even in an immunocompetent patient, and we suggest a genotypic assay, such as 16S rRNA sequencing, to confirm species identity. PMID:27659439

  7. Effects of bovine subclinical mastitis caused by Corynebacterium spp. on somatic cell count, milk yield and composition by comparing contralateral quarters.

    Science.gov (United States)

    Gonçalves, Juliano Leonel; Tomazi, Tiago; Barreiro, Juliana Regina; Beuron, Daniele Cristine; Arcari, Marcos André; Lee, Sarah Hwa In; Martins, Cristian Marlon de Magalhães Rodrigues; Araújo Junior, João Pessoa; dos Santos, Marcos Veiga

    2016-03-01

    Subclinical mastitis caused by Corynebacterium spp. (as a group and at the species level) was investigated by evaluating contralateral (healthy and infected) mammary quarters for somatic cell count (SCC), milk yield and composition. Selection of cows with subclinical mastitis caused by Corynebacterium spp. was performed by microbiological culture of composite samples collected from 1242 dairy cows from 21 dairy herds. For each of the selected cows, milk yield was measured and milk samples were collected at the mammary quarter level (i.e., 1140 mammary samples collected from 285 cows) for analysis of milk composition and SCC. The identification of Corynebacterium spp. isolates was performed by 16S rRNA gene sequencing. One hundred and eighty Corynebacterium spp. isolates were identified, of which 167 (92.77%) were C.bovis and eight (4.44%) non-C.bovis; for five of the Corynebacterium spp. isolates (2.77%), sequencing of 16S rRNA genes did not allow identification at the species level. Mammary quarters infected with Corynebacterium spp. as a group had a higher geometric mean SCC (197,900 cells/mL) than healthy contralateral mammary quarters (85,800 cells/mL). Species of Corynebacterium non-C.bovis were infrequently isolated and did not change SCC, milk yield or milk solid contents when evaluated at the contralateral quarter level. Although C.bovis infection showed no effect on milk yield, fat, protein, casein or total solids in milk, it increased SCC and decreased lactose and milk solids non-fat content. PMID:26831159

  8. Biochemical and molecular characterization of the gentisate transporter GenK in Corynebacterium glutamicum.

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    Ying Xu

    Full Text Available BACKGROUND: 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 [(14C]-labeled gentisate has not been previously reported. METHODOLOGY/PRINCIPAL FINDINGS: In this study, biochemical characterization of GenK by uptake assays with [(14C]-labeled substrates demonstrated that it specifically transported gentisate into the cells with V(max and K(m of 3.06 ± 0.16 nmol/min/mg of dry weight and 10.71 ± 0.11 µM respectively, and no activity was detected for either benzoate or 3-hydoxybenzoate. When GenK was absent in strain RES167 ΔgenK, it retained 85% of its original transport activity at pH 6.5 compared to that of strain RES167. However, it lost 79% and 88% activity at pH 7.5 and 8.0, respectively. A number of competing substrates, including 3-hydroxybenzoate, benzoate, protocatechuate and catechol, significantly inhibited gentisate uptake by more than 40%. Through site-directed mutagenesis, eight amino acid residues of GenK, Asp-54, Asp-57 and Arg-386 in the hydrophobic transmembrane regions and Arg-103, Trp-309, Asp-312, Arg-313 and Ile-317 in the hydrophilic cytoplasmic loops were shown to be important for gentisate transport. When conserved residues Asp-54 and Asp-57 respectively were changed to glutamate, both mutants retained approximately 50% activity and were able to partially complement the ability of strain RES167 ΔgenK to grow on gentisate. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that GenK is an active gentisate transporter in Corynebacterium glutamicum ATCC13032. The GenK-mediated gentisate transport was also shown to be a limiting step for the gentisate utilization by this strain. This enhances our

  9. Epidemiological survey of Corynebacterium equi infections on five Ontario horse farms.

    Science.gov (United States)

    Prescott, J F; Travers, M; Yager-Johnson, J A

    1984-01-01

    Corynebacterium equi was cultured from manure or soil on five horse-breeding farms in Ontario at monthly intervals on three occasions during the summer of 1982. The organism was widespread. Contamination by C. equi of the loafing paddock and pasture areas was significantly greater in a farm established 30 years than in two established for four and six years and there was a significant correlation between the C. equi burden in stables, paddocks and pastures and the length of use of the five farms for horses. In all farms, numbers of C. equi in pasture soil exceeded numbers in fresh manure, suggesting that environmental multiplication of the organism might occur. A farm with an endemic C. equi pneumonia problem differed significantly from the other four farms, where disease was not endemic, in the larger number of C. equi isolated in the stable area. By contrast the farm with a C. equi pasture soil burden significantly heavier than on all other farms had no deaths due to C. equi pneumonia. There was a correlation (r = 0.78, p = 0.061) between the number of cases of C. equi pneumonia on the farms and numbers of C. equi in the area of the stables, but not on the paddocks or pastures. About two-thirds of randomly chosen isolates from the farms belonged to the three capsular serotypes most commonly found in pneumonic foals.

  10. Engineering of Corynebacterium glutamicum for growth and succinate production from levoglucosan, a pyrolytic sugar substrate.

    Science.gov (United States)

    Kim, Eun-Mi; Um, Youngsoon; Bott, Michael; Woo, Han Min

    2015-10-01

    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 Pseudomonas aeruginosa was employed for levoglucosan conversion in C. glutamicum because the wild type was unable to utilize levoglucosan as sole carbon source. As result, expression of a levoglucosan kinase (LGK) of L. starkeyi only enabled growth with levoglucosan as sole carbon source in CgXII minimal medium by catalyzing conversion of levoglucosan to glucose-6-phosphate. Subsequently, the lgk gene was expressed in an aerobic succinate producer of C. glutamicum, strain BL-1. The recombinant strain showed a higher succinate yield (0.25 g g(-1)) from 2% (w/v) levoglucosan than the reference strain BL-1 from 2% (w/v) glucose (0.19 g g(-1)), confirming that levoglucosan is an attractive carbon substrate for C. glutamicum producer strains. In summary, we demonstrated that a pyrolytic sugar could be a potential carbon source for microbial cell factories. PMID:26363018

  11. Metabolic engineering of Corynebacterium glutamicum strain ATCC13032 to produce L-methionine.

    Science.gov (United States)

    Qin, Tianyu; Hu, Xiaoqing; Hu, Jinyu; Wang, Xiaoyuan

    2015-01-01

    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 feedback-resistant aspartate kinase and the gene hom(m) encoding feedback-resistant homoserine dehydrogenase to further increase the precursor supply, and (iv) overexpression of the gene cluster brnF and brnE encoding the export protein complex BrnFE to increase extracellular l-methionine concentration. QW102/pJYW-4-hom(m) -lysC(m) -brnFE produced 42.2 mM (6.3 g/L) l-methionine after 64-H fed-batch fermentation. These results suggest that l-methionine-producing strains can be developed from wild-type C. glutamicum strains by rationally metabolic engineering.

  12. Corynebacterium pseudotuberculosis Infection (Caseous Lymphadenitis in Camels (Camelus dromedarius in Jordan

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    Azmi D. Hawari

    2008-01-01

    Full Text Available Problem statement: This study was conducted to describe & report for the first time outbreaks of natural C.pseudotuberculosis infection in adult camel herds (Camelus dromedarius in Jordan. An infectious disease syndrome was reported in three camel herds (Camelus dromedarius intensively raised at south province in Jordan. Approach: The herds included over 160 adult camels out of which about 8% were affected with multiple muscle and subcutaneous abscesses at various sites of the body. The camels were also heavily infested with ticks. Results: The infected camels did not respond favorably to several broad spectrum antibiotics. Post-mortem examination of 5 carcasses revealed emaciation and presence of external and internal multiple abscesses particularly in the lungs. The abscesses were encapsulated by fibrous tissue and contained creamy yellowish white pus. The lymph nodes were slightly congested and swollen. Conclusion: Corynebacterium pseudotuberculosis type I strain or biovar ovis (the known cause of caseous lymphadenitis in sheep was isolated from pus, lymph nodes, ticks, milk, blood and liver samples. The clinical symptoms, nature and distribution of lesions of caseous lymphadenitis in camels are not as typical as in sheep. Recommendations for pseudotuberculosis control were given.

  13. Surgical Site Infection by Corynebacterium macginleyi in a Patient with Neurofibromatosis Type 1

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    Bruno Cacopardo

    2013-01-01

    Full Text Available Corynebacterium (C. macginleyi is a gram positive, lipophilic rod, usually considered a colonizer of skin and mucosal surfaces. Several reports have associated C. macginleyi with ocular infections, such as conjunctivitis and endophthalmitis. However, even if rare, extraocular infections from C. macginleyi may occur, especially among immunocompromised patients and patients with indwelling medical devices. We report herein the first case of surgical site infection by C. macginleyi after orthopaedic surgery for the correction of kyphoscoliosis in a patient with neurofibromatosis type 1. Our patient developed a nodular granulomatous lesion of about two centimetres along the surgical scar, at the level of C4-C5, with purulent discharge and formation of a fistulous tract. Cervical magnetic resonance imaging showed the presence of a two-centimetre fluid pocket in the subcutaneous tissue. Several swabs were collected from the borders of the lesion as well as from the exudate, with isolation of C. macginleyi. The isolate was susceptible to beta-lactams, cotrimoxazole, linezolid, and glycopeptides but resistant to quinolones, third-generation cephalosporins, and erythromycin. Two 30-day courses of antibiotic therapy with amoxicillin/clavulanate (1 g three times/day and cotrimoxazole (800/160 mg twice a day were administered, obtaining a complete healing of the lesion.

  14. Production of L-ornithine from sucrose and molasses by recombinant Corynebacterium glutamicum.

    Science.gov (United States)

    Zhang, Yuan-Yuan; Bu, Yi-Fan; Liu, Jian-Zhong

    2015-09-01

    Sucrose and molasses are attractive raw materials for industrial fermentation. Although Corynebacterium glutamicum shows sucrose-utilizing activity, sucrose or molasses is only a fraction of carbon source used in the fermentation medium in most works. An engineered C. glutamicum strain was constructed for producing L-ornithine with sucrose or molasses as a sole carbon source by transferring Mannheimia succiniciproducens β-fructofuranosidase gene (sacC). The engineered strain, C. glutamicum ΔAPE6937R42 (pEC-sacC), produced 22.0 g/L of L-ornithine with sucrose as the sole carbon source, which is on par with that obtained by the parent strain C. glutamicum ΔAPE6937R42 with glucose as the sole carbon. The resulting strain C. glutamicum ΔAPE6937R42 (pEC-sacC) produced 27.0 g/L of L-ornithine with molasses as the sole carbon source, which is higher than that obtained by the parent strain C. glutamicum ΔAPE6937R42 with glucose as the sole carbon. This strategy can be applied for developing sucrose- or molasses-utilizing industrial strains.

  15. The amrG1 gene is involved in the activation of acetate in Corynebacterium glutamicum

    Institute of Scientific and Technical Information of China (English)

    RUAN Hong; R. Gerstmeir; S. Schnicke; B.J. Eikmanns

    2005-01-01

    During growth of Corynebacterium glutamicum on acetate as its carbon and energy source, the expression of the pta-ack operon is induced, coding for the acetate-activating enzymes, which are phosphotransacetylase (PTA) and acetate kinase (AK). By transposon rescue, we identified the two genes amrG1 and amrG2 found in the deregulated transposon mutant C. glutamicum G25. The amrG1 gene (NCBI-accession: AF532964) has a size of 732 bp, encoding a polypeptide of 243 amino acids and apparently is partially responsible for the regulation of acetate metabolism in C. glutamicum. We constructed an in-frame deletion mutant and an overexpressing strain of amrG1 in the C. glutamicum ATCC13032 wildtype. The strains were then analyzed with respect to their enzyme activities of PTA and AK during growth on glucose, acetate and glucose or acetate alone as carbon sources. Compared to the parental strain, the amrG1 deletion mutant showed higher specific AK and PTA activities during growth on glucose but showed the same high specific activities of AK and PTA on medium containing acetate plus glucose and on medium containing acetate. In contrast to the gene deletion, overexpression of the amrG1 gene in C. glutamicum 13032 had the adverse regulatory effect. These results indicate that the amrG1 gene encodes a repressor or co-repressor of the pta-ack operon.

  16. Teste de pele em caprinos vacinados e infectados com Corynebacterium pseudotuberculosis

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    Francisco Selmo Fernandes Alves

    1999-07-01

    Full Text Available Dez caprinos foram vacinados com toxóide a 3%, outros dez com uma bacterina e mais dois grupos-controle de cinco animais cada, submetidos à inoculação de infusão de cérebro e coração e solução salina, respectivamente. Todos os animais foram examinados e avaliados com um teste de pele. Tanto o toxóide quanto a bacterina foram produzidos a partir de amostra de Corynebacterium pseudotuberculosis. Todos os caprinos foram desafiados com C. pseudotuberculosis, trinta dias após as vacinações. Nenhuma das vacinas induziu reação de hipersensibilidade na pele dos caprinos antes do desafio. Após o desafio, todos os animais desenvolveram reações mensuráveis na primeira, quinta e décima semana em resposta ao teste de pele. Os diâmetros da reação dérmica aumentaram do décimo dia à quinta semana após o desafio. As medidas alcançaram tamanho maior na décima semana. O resultado deste estudo indica que antígeno específico do C. pseudotuberculosis pode ser utilizado em caprinos no diagnóstico da linfadenite caseosa como teste de pele ou como instrumento experimental para monitorar o desenvolvimento da doença.

  17. Septicaemia secondary to infection by Corynebacterium macginleyi in an Indian python (Python molurus).

    Science.gov (United States)

    Martínez, Jorge; Segura, Pablo; García, David; Aduriz, Gorka; Ibabe, José C; Peris, Bernardo; Corpa, Juan M

    2006-09-01

    A seven-year-old female Indian python (Python molurus) weighing about 35kg was euthanased after several clinical episodes of stomatitis, pneumonia, ophthalmitis and dystocia over a period of four years. The animal had been maintained in a terrarium in a circus truck at an adequate temperature. During shows, however, the snake was considered to be exposed to stressful conditions for several hours at a time at low temperatures and with noise and bright lights. A post-mortem examination indicated ulcerative stomatitis, osteomyelitis, severe pneumonia and numerous granulomata and multifocal necrosis in stomach and spleen. Corynebacterium macginleyi was isolated in pure culture from the ulcerative stomatitis, and mixed with Stenotrophomonas maltophilia from the lungs and spleen. The findings indicated that the snake had died from a septicaemic process caused by C. macginleyi, probably originating from the stomatitis. The role of S. maltophilia as a secondary agent is discussed. The stress of the circus show and poor husbandry may have predisposed the animal to infection and septicaemia. This is the first report of C. macginleyi causing disease in a snake.

  18. Study of Corynebacterium diphtheriae strains isolated in Romania, northwestern Russia and the Republic of Moldova.

    Science.gov (United States)

    Damian, Maria; Grimont, Francine; Narvskaya, Olga; Straut, Monica; Surdeanu, Maria; Cojocaru, Radu; Mokrousov, Igor; Diaconescu, Angela; Andronescu, Constantin; Melnic, Anatol; Mutoi, Ludmila; Grimont, Patrick A D

    2002-03-01

    A selection of 167 Corynebacterium diphtheriae strains isolated in Romania, the Russian Federation and the Republic of Moldova were analysed by biotyping, phage typing, the toxin production test and by molecular techniques such as ribotyping, pulsed field gel electrophoresis and random amplified polymorphic DNA, in order to establish the epidemiological relatedness, genetic divergence and strain circulation within and between the bordering countries. Using a set of five digoxigenin-labeled oligonucleotides and BstEII digestion, 34 ribotypes were identified. The strains isolated in the epidemic areas (Russia and Moldova) were very closely related but different from those isolated in Romania. C1 and C5 were the main ribotypes identified in these areas. Neither ribotype was found in Romania, where the main circulating types were C3 and C7. Field inversion gel electrophoresis was more discriminative than ribotyping and revealed 54 macrorestriction profiles after SfiI restriction. Both methods showed a significant homogeneity of the strains from epidemic areas and a large diversity among the Romanian strains. Random amplification was useful as an identification method for the epidemic strains, but not for the Romanian ones which displayed a large number of amplification profiles. The phenotypic methods associated with molecular typing techniques enabled distinguishing between strains, detecting the epidemic clone, and sustaining the absence of transmission across borders.

  19. Epidemiological survey of Corynebacterium equi infections on five Ontario horse farms.

    Science.gov (United States)

    Prescott, J F; Travers, M; Yager-Johnson, J A

    1984-01-01

    Corynebacterium equi was cultured from manure or soil on five horse-breeding farms in Ontario at monthly intervals on three occasions during the summer of 1982. The organism was widespread. Contamination by C. equi of the loafing paddock and pasture areas was significantly greater in a farm established 30 years than in two established for four and six years and there was a significant correlation between the C. equi burden in stables, paddocks and pastures and the length of use of the five farms for horses. In all farms, numbers of C. equi in pasture soil exceeded numbers in fresh manure, suggesting that environmental multiplication of the organism might occur. A farm with an endemic C. equi pneumonia problem differed significantly from the other four farms, where disease was not endemic, in the larger number of C. equi isolated in the stable area. By contrast the farm with a C. equi pasture soil burden significantly heavier than on all other farms had no deaths due to C. equi pneumonia. There was a correlation (r = 0.78, p = 0.061) between the number of cases of C. equi pneumonia on the farms and numbers of C. equi in the area of the stables, but not on the paddocks or pastures. About two-thirds of randomly chosen isolates from the farms belonged to the three capsular serotypes most commonly found in pneumonic foals. PMID:6713248

  20. Metabolic engineering of Corynebacterium glutamicum ATCC13869 for L-valine production.

    Science.gov (United States)

    Chen, Cheng; Li, Yanyan; Hu, Jinyu; Dong, Xunyan; Wang, Xiaoyuan

    2015-05-01

    In this study, an L-valine-producing strain was developed from Corynebacterium glutamicum ATCC13869 through deletion of the three genes aceE, alaT and ilvA combined with the overexpression of six genes ilvB, ilvN, ilvC, lrp1, brnF and brnE. Overexpression of lrp1 alone increased L-valine production by 16-fold. Deletion of the aceE, alaT and ilvA increased L-valine production by 44-fold. Overexpression of the six genes ilvB, ilvN, ilvC, lrp1, brnE and brnF in the triple deletion mutant WCC003 further increased L-valine production. The strain WCC003/pJYW-4-ilvBNC1-lrp1-brnFE produced 243mM L-valine in flask cultivation and 437mM (51g/L) L-valine in fed-batch fermentation and lacked detectable amino-acid byproduct such as l-alanine and l-isoleucine that are usually found in the fermentation of L-valine-producing C. glutamicum.

  1. Biosensor-driven adaptive laboratory evolution of l-valine production in Corynebacterium glutamicum.

    Science.gov (United States)

    Mahr, Regina; Gätgens, Cornelia; Gätgens, Jochem; Polen, Tino; Kalinowski, Jörn; Frunzke, Julia

    2015-11-01

    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-responsive sensor based on the transcriptional regulator Lrp of C. glutamicum. Evolved strains featured a significantly higher growth rate, increased L-valine titers (~25%) and a 3-4-fold reduction of by-product formation. Genome sequencing resulted in the identification of a loss-of-function mutation (UreD-E188*) in the gene ureD (urease accessory protein), which was shown to increase L-valine production by up to 100%. Furthermore, decreased L-alanine formation was attributed to a mutation in the global regulator GlxR. These results emphasize biosensor-driven evolution as a straightforward approach to improve growth and productivity of microbial production strains.

  2. Heterologous expression and localization of gentisate transporter Ncg12922 from Corynebacterium glutamicum ATCC 13032

    International Nuclear Information System (INIS)

    Ralstonia sp. strain U2 metabolizes naphthalene via gentisate (2,5-dihydroxybenzoate) to central metabolites, but it was found unable to utilize gentisate as growth substrate. A putative gentisate transporter encoded by ncg12922 from Corynebacterium glutamicum ATCC 13032 was functionally expressed in Ralstonia sp. strain U2, converting strain U2 to a gentisate utilizer. After ncg12922 was inserted into plasmid pGFPe with green fluorescence protein gene gfp, the expressed fusion protein Ncg12922-GFP could be visualized in the periphery of Escherichia coli cells under confocal microscope, consistent with a cytoplasmic membrane location. In contrast, GFP was ubiquitous in the cytoplasm of E. coli cells carrying pGFPe only. Gentisate 1,2-dioxygenase activity was present in the cell extract from strain U2 induced with gentisate but at a much lower level (one-fifth) than that obtained with salicylate. However, it exhibited a similar level in strain U2 containing Ncg12922 induced either by salicylate or gentisate

  3. The small 6C RNA of Corynebacterium glutamicum is involved in the SOS response.

    Science.gov (United States)

    Pahlke, Jennifer; Dostálová, Hana; Holátko, Jiří; Degner, Ursula; Bott, Michael; Pátek, Miroslav; Polen, Tino

    2016-09-01

    The 6C RNA family is a class of small RNAs highly conserved in Actinobacteria, including the genera Mycobacterium, Streptomyces and Corynebacterium whose physiological function has not yet been elucidated. We found that strong transcription of the cgb_03605 gene, which encodes 6C RNA in C. glutamicum, was driven by the SigA- and SigB-dependent promoter Pcgb_03605. 6C RNA was detected at high level during exponential growth phase (180 to 240 molcules per cell) which even increased at the entry of the stationary phase. 6C RNA level did not decrease within 240 min after transcription had been stopped with rifampicin, which suggests high 6C RNA stability. The expression of cgb_03605 further increased approximately twofold in the presence of DNA-damaging mitomycin C (MMC) and nearly threefold in the absence of LexA. Deletion of the 6C RNA gene cgb_03605 resulted in a higher sensitivity of C. glutamicum toward MMC and UV radiation. These results indicate that 6C RNA is involved in the DNA damage response. Both 6C RNA level-dependent pausing of cell growth and branched cell morphology in response to MMC suggest that 6C RNA may also be involved in a control of cell division.

  4. Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum

    Institute of Scientific and Technical Information of China (English)

    CHEN Xue Lan; ZHANG Bin; TANG Li; JIAO Hai Tao; XU Heng Yi; XU Feng; XU Hong; WEI Hua; XIONG Yong Hua

    2014-01-01

    Objective Corynebacterium crenatum MT, a mutant from C. crenatum AS 1.542 with a lethal argR gene, exhibits high arginine production. To confirm the effect of ArgR on arginine biosynthesis in C. crenatum, an intact argR gene from wild-type AS 1.542 was introduced into C. crenatum MT, resulting in C. crenatum MT. sp, and the changes of transcriptional levels of the arginine biosynthetic genes and arginine production were compared between the mutant strain and the recombinant strain. Methods Quantitative real-time polymerase chain reaction was employed to analyze the changes of the related genes at the transcriptional level, electrophoretic mobility shift assays were used to determine ArgR binding with the argCJBDF, argGH, and carAB promoter regions, and arginine production was determined with an automated amino acid analyzer. Results Arginine production assays showed a 69.9%reduction in arginine from 9.01±0.22 mg/mL in C. crenatum MT to 2.71±0.13 mg/mL (P Conclusion The arginine biosynthetic genes in C. crenatum are clearly controlled by the negative regulator ArgR, and intact ArgR in C. crenatum MT results in a significant descrease in arginine production.

  5. Combined radiotherapy and Corynebacterium parvum treatment of rat tumors with different immunogenicity. [X rays

    Energy Technology Data Exchange (ETDEWEB)

    Moroson, H.; Stowe, S.; Rotman, M.; Schechter, M.

    1978-01-01

    Evidence is presented that combined radiotherapy and Corynebacterium parvum treatment gives better results than radiotherapy alone in rats bearing a chemically-induced highly-immunogenic transplanted fibrosarcoma termed BP 179; however, similar behavior is not observed with either of two weakly-immunogenic mammary carcinomas, 13762 or ME/H. Relative immunogenicity is determined by the ability of immunized rats to reject tumor cell challenge. Both 13762 and ME/H carcinomata grow progressively and metastasize early to the retroperitoneal cavity and lungs if they are left untreated. Local radiotherapy of the primary tumor has no influence on growth of metastases whether it is combined with C. parvum or not. Results of cell-mediated cytotoxicity studies with lymphocytes from BP 179 and ME/H tumor bearing rats treated with radiation or radiation plus C. parvum support the in vivo findings of combined radiotherapy. These data suggest that unlike strongly immunogenic tumors, weakly immunogenic tumors will not respond better to C. parvum combined with radiation therapy.

  6. Mechanism for macrophage activation against Corynebacterium parvum--participation of T cells and its lymphokines.

    Science.gov (United States)

    Mori, H; Mihara, M; Uesugi, Y; Nagai, H; Koda, A

    1994-01-01

    It is well known that Corynebacterium parvum activates macrophages to produce tumor necrosis factor (TNF). It is suspected that the activation of macrophages by C. parvum requires T-cell participation. The purpose of this study was to confirm that T cells participate in the activation of macrophages by C. parvum. TNF production in vitro from the spleen cells of BALB/c(-)+/+ mice was abrogated completely by the pre-treatment of spleen cells with anti-Ia antiserum and complement, indicating that Ia+ cells are the source of TNF. TNF production was not elicited at all in BALB/c-nu/nu mice. However, there was an increase in the number of Ia+ cells as well as an increase in the weight of spleen and liver. Supernatant from a culture of spleen cells stimulated with phytohemagglutinin-P (a PHA-induced lymphokine) made it possible for BALB/c-nu/nu mice to produce TNF, associated with an induction of Lyt-1+ cells and Lyt-2+ cells. However, treatment with the lymphokine did not augment the increases of Ia+ cells or liver and spleen weights. These results suggest that increasing the number of Ia+ cells is not sufficient to bring about TNF production; Ia+ cells must also be stimulated by T cells or T-cell lymphokines in order to produce TNF. These results suggest that T cells play an essential role in the activation of Ia+ cells against C. parvum. PMID:7723692

  7. Seroma prevention by using Corynebacterium parvum in a rat mastectomy model.

    Science.gov (United States)

    Tekin, E; Kocdor, M A; Saydam, S; Bora, S; Harmancioglu, O

    2001-01-01

    Seroma formation is the most common complication after mastectomy and continues to be an important problem during the early postoperative period. Several surgical and medical methods have been developed to try to overcome this problem; however, so far none have been used successfully in the routine clinical practice. The aim of this study is to evaluate the effects of Corynebacterium parvum (CP) as a sclerosing agent in both prevention and treatment of seromas after mastectomy and axillary dissection in an animal model. Sixty female Sprague-Dawley rats underwent mastectomy and axillary dissection under general anaesthesia. Following surgery, the rats were treated in 1 of 3 ways. In the prevention group, 1 cm3 (0.35 mg) CP solution was injected beneath the skin flap just before closure of the incision after mastectomy. In the treatment group, animals in which a seroma was formed, the fluid was aspirated, and 1 cm3 CP solution was injected beneath the flap. In the control group, animals in which seromas formed, aspiration only was performed. The frequency of seroma formation decreased when CP solution was injected immediately after the operation (p treatment group in this experimental model. CP injection may be useful for the management of this problem in a clinical setting. PMID:11490129

  8. Biosensor-driven adaptive laboratory evolution of l-valine production in Corynebacterium glutamicum.

    Science.gov (United States)

    Mahr, Regina; Gätgens, Cornelia; Gätgens, Jochem; Polen, Tino; Kalinowski, Jörn; Frunzke, Julia

    2015-11-01

    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-responsive sensor based on the transcriptional regulator Lrp of C. glutamicum. Evolved strains featured a significantly higher growth rate, increased L-valine titers (~25%) and a 3-4-fold reduction of by-product formation. Genome sequencing resulted in the identification of a loss-of-function mutation (UreD-E188*) in the gene ureD (urease accessory protein), which was shown to increase L-valine production by up to 100%. Furthermore, decreased L-alanine formation was attributed to a mutation in the global regulator GlxR. These results emphasize biosensor-driven evolution as a straightforward approach to improve growth and productivity of microbial production strains. PMID:26453945

  9. Characterization of citrate utilization in Corynebacterium glutamicum by transcriptome and proteome analysis.

    Science.gov (United States)

    Polen, Tino; Schluesener, Daniela; Poetsch, Ansgar; Bott, Michael; Wendisch, Volker F

    2007-08-01

    Corynebacterium glutamicum grows aerobically on a variety of carbohydrates and organic acids as single or combined sources of carbon and energy. To characterize the citrate utilization in C. glutamicum on a genomewide scale, a comparative analysis was carried out by combining transcriptome and proteome analysis. In cells grown on citrate, transcriptome analysis revealed highest expression changes for two different citrate-uptake systems encoded by citM and tctCBA, whereas genes encoding uptake systems for the glucose- (ptsG), sucrose- (ptsS) and fructose- (ptsF) specific PTS components and permeases for gluconate (gntP) and glutamate (gluC) displayed decreased mRNA levels in citrate-grown cells. This pattern was also observed when cells grown in Luria-Bertani (LB) medium plus citrate were compared with cells grown in LB medium, indicating some kind of catabolite repression. Genes encoding enzymes of the tricarboxylic acid cycle (aconitase, succinyl-CoA synthetase, succinate dehydrogenase and fumarase), malic enzyme, PEP carboxykinase, gluconeogenic glyceraldehyde-3-phosphate dehydrogenase and ATP synthase displayed increased expression in cells grown on citrate. Accordingly, proteome analysis revealed elevated protein levels of these enzymes and showed a good correlation with the mRNA levels. In conclusion, this study revealed the citrate stimulon in C. glutamicum and the regulated central metabolic genes when grown on citrate. PMID:17559405

  10. Pupylated proteins in Corynebacterium glutamicum revealed by MudPIT analysis.

    Science.gov (United States)

    Küberl, Andreas; Fränzel, Benjamin; Eggeling, Lothar; Polen, Tino; Wolters, Dirk Andreas; Bott, Michael

    2014-06-01

    In a manner similar to ubiquitin, the prokaryotic ubiquitin-like protein (Pup) has been shown to target proteins for degradation via the proteasome in mycobacteria. However, not all actinobacteria possessing the Pup protein also contain a proteasome. In this study, we set out to study pupylation in the proteasome-lacking non-pathogenic model organism Corynebacterium glutamicum. A defined pup deletion mutant of C. glutamicum ATCC 13032 grew aerobically as the parent strain in standard glucose minimal medium, indicating that pupylation is dispensable under these conditions. After expression of a Pup derivative carrying an aminoterminal polyhistidine tag in the Δpup mutant and Ni(2+)-chelate affinity chromatography, pupylated proteins were isolated. Multidimensional protein identification technology (MudPIT) and MALDI-TOF-MS/MS of the elution fraction unraveled 55 proteins being pupylated in C. glutamicum and 66 pupylation sites. Similar to mycobacteria, the majority of pupylated proteins are involved in metabolism or translation. Our results define the first pupylome of an actinobacterial species lacking a proteasome, confirming that other fates besides proteasomal degradation are possible for pupylated proteins. PMID:24737727

  11. The small 6C RNA of Corynebacterium glutamicum is involved in the SOS response.

    Science.gov (United States)

    Pahlke, Jennifer; Dostálová, Hana; Holátko, Jiří; Degner, Ursula; Bott, Michael; Pátek, Miroslav; Polen, Tino

    2016-09-01

    The 6C RNA family is a class of small RNAs highly conserved in Actinobacteria, including the genera Mycobacterium, Streptomyces and Corynebacterium whose physiological function has not yet been elucidated. We found that strong transcription of the cgb_03605 gene, which encodes 6C RNA in C. glutamicum, was driven by the SigA- and SigB-dependent promoter Pcgb_03605. 6C RNA was detected at high level during exponential growth phase (180 to 240 molcules per cell) which even increased at the entry of the stationary phase. 6C RNA level did not decrease within 240 min after transcription had been stopped with rifampicin, which suggests high 6C RNA stability. The expression of cgb_03605 further increased approximately twofold in the presence of DNA-damaging mitomycin C (MMC) and nearly threefold in the absence of LexA. Deletion of the 6C RNA gene cgb_03605 resulted in a higher sensitivity of C. glutamicum toward MMC and UV radiation. These results indicate that 6C RNA is involved in the DNA damage response. Both 6C RNA level-dependent pausing of cell growth and branched cell morphology in response to MMC suggest that 6C RNA may also be involved in a control of cell division. PMID:27362471

  12. Degradation and assimilation of aromatic compounds by Corynebacterium glutamicum: another potential for applications for this bacterium?

    Science.gov (United States)

    Shen, Xi-Hui; Zhou, Ning-Yi; Liu, Shuang-Jiang

    2012-07-01

    With the implementation of the well-established molecular tools and systems biology techniques, new knowledge on aromatic degradation and assimilation by Corynebacterium glutamicum has been emerging. This review summarizes recent findings on degradation of aromatic compounds by C. glutamicum. Among these findings, the mycothiol-dependent gentisate pathway was firstly discovered in C. glutamicum. Other important knowledge derived from C. glutamicum would be the discovery of linkages among aromatic degradation and primary metabolisms such as gluconeogenesis and central carbon metabolism. Various transporters in C. glutamicum have also been identified, and they play an essential role in microbial assimilation of aromatic compounds. Regulation on aromatic degradation occurs mainly at transcription level via pathway-specific regulators, but global regulator(s) is presumably involved in the regulation. It is concluded that C. glutamicum is a very useful model organism to disclose new knowledge of biochemistry, physiology, and genetics of the catabolism of aromatic compounds in high GC content Gram-positive bacteria, and that the new physiological properties of aromatic degradation and assimilation are potentially important for industrial applications of C. glutamicum.

  13. Crystallization and preliminary X-ray diffraction studies of FAD synthetase from Corynebacterium ammoniagenes

    International Nuclear Information System (INIS)

    Native and selenomethionine-labelled FAD synthetase from C. ammoniagenes have been crystallized by the hanging-drop vapour-diffusion method. A MAD data set for SeMet-labelled FAD synthetase was collected to 2.42 Å resolution, while data sets were collected to 1.95 Å resolution for the native crystals. FAD synthetase from Corynebacterium ammoniagenes (CaFADS), a prokaryotic bifunctional enzyme that catalyses the phosphorylation of riboflavin as well as the adenylylation of FMN, has been crystallized using the hanging-drop vapour-diffusion method at 277 K. Diffraction-quality cubic crystals of native and selenomethionine-labelled (SeMet-CaFADS) protein belonged to the cubic space group P213, with unit-cell parameters a = b = c = 133.47 Å and a = b = c = 133.40 Å, respectively. Data sets for native and SeMet-containing crystals were collected to 1.95 and 2.42 Å resolution, respectively

  14. Evolution, epidemiology and diversity of Corynebacterium diphtheriae: New perspectives on an old foe.

    Science.gov (United States)

    Sangal, Vartul; Hoskisson, Paul A

    2016-09-01

    Diphtheria is a debilitating disease caused by toxigenic Corynebacterium diphtheriae strains and has been effectively controlled by the toxoid vaccine, yet several recent outbreaks have been reported across the globe. Moreover, non-toxigenic C. diphtheriae strains are emerging as a major global health concern by causing severe pharyngitis and tonsillitis, endocarditis, septic arthritis and osteomyelitis. Molecular epidemiological investigations suggest the existence of outbreak-associated clones with multiple genotypes circulating around the world. Evolution and pathogenesis appears to be driven by recombination as major virulence factors, including the tox gene and pilus gene clusters, are found within genomic islands that appear to be mobile between strains. The number of pilus gene clusters and variation introduced by gain or loss of gene function correlate with the variable adhesive and invasive properties of C. diphtheriae strains. Genomic variation does not support the separation of C. diphtheriae strains into biovars which correlates well with findings of studies based on multilocus sequence typing. Genomic analyses of a relatively small number of strains also revealed a recombination driven diversification of strains within a sequence type and indicate a wider diversity among C. diphtheriae strains than previously appreciated. This suggests that there is a need for increased effort from the scientific community to study C. diphtheriae to help understand the genomic diversity and pathogenicity within the population of this important human pathogen. PMID:27291708

  15. Reproductive Pathological Changes Associated with Experimental Subchronic Corynebacterium pseudotuberculosis Infection in Nonpregnant Boer Does

    Directory of Open Access Journals (Sweden)

    A. M. Othman

    2016-01-01

    Full Text Available Corynebacterium pseudotuberculosis causes caseous lymphadenitis (CLA, which is a contagious and chronic disease in sheep and goats. In order to assess the histopathological changes observed in the reproductive organs of nonpregnant does infected with the bacteria, 20 apparently healthy adult Boer does were divided into four inoculation groups, intradermal, intranasal, oral, and control, consisting of five goats each. Excluding the control group, which was unexposed, other does were inoculated with 107 CFU/1 mL of live C. pseudotuberculosis through the various routes stated above. Thirty days after infection, the ovaries, uterus, and iliac lymph nodes were collected for bacterial recovery and molecular detection, as well as histopathological examination. The mean changes in necrosis, congestion, inflammatory cell infiltration, and oedema varied in severity among the ovaries, uterus, and iliac lymph nodes following different inoculation routes. Overall, the intranasal route of inoculation showed more severe (p<0.05 lesions in all the organs examined. The findings of this study have shown that C. pseudotuberculosis could predispose to infertility resulting from pathological lesions in the uterus and ovaries of does.

  16. In silico identification of Corynebacterium pseudotuberculosis antigenic targets and application in immunodiagnosis.

    Science.gov (United States)

    Rezende, Andrea de Fátima Silva; Brum, Alexandre Antunes; Reis, Carlos Guilherme; Angelo, Henrique Ramos; Leal, Karen Silva; Silva, Mara Thais de Oliveira; Simionatto, Simone; Azevedo, Vasco; Santos, Anderson; Portela, Ricardo Wagner; Dellagostin, Odir; Borsuk, Sibele

    2016-06-01

    Caseous lymphadenitis (CLA) is a disease caused by Corynebacterium pseudotuberculosis. It affects mainly small ruminants and causes significant economic losses worldwide. Because symptoms are not immediately noticeable, CLA clinical diagnosis is not effective. Numerous serological tests are being developed to detect the disease in asymptomatic animals, but currently available immunoassays have problems with sensitivity. Current ELISA formats use native bacterial antigens, and recombinant proteins could be useful for improving the immunoassay parameters. The C. pseudotuberculosis proteins CP0126a, CP0369 and CP1957 were identified from 2097 candidate proteins by mature epitope density (MED) analysis, expressed in Escherichia coli and evaluated in an indirect immunoenzymic system. The CP0126a, CP0369 and CP1957 ELISAs showed 77.5 %, 92.5 % and 92.5 % specificity and 95 %, 90 % and 85 % sensitivity, respectively. Receiver operating characteristic (ROC) curve analysis showed an area under the curve of 0.874, 0.951 and 0.881, respectively. The proteins identified in silico were recognized by antibodies in the sera from infected animals without being recognized in negative samples. The ELISA assay using the rCP0369 protein as antigen had the greatest specificity and sensitivity values, followed by rCP1957. This is an interesting strategy for seroepidemiological investigations in sheep flocks due to its significant specificity and sensitivity. PMID:27071381

  17. Characterization of a Unique Pathway for 4-Cresol Catabolism Initiated by Phosphorylation in Corynebacterium glutamicum.

    Science.gov (United States)

    Du, Lei; Ma, Li; Qi, Feifei; Zheng, Xianliang; Jiang, Chengying; Li, Ailei; Wan, Xiaobo; Liu, Shuang-Jiang; Li, Shengying

    2016-03-18

    4-Cresol is not only a significant synthetic intermediate for production of many aromatic chemicals, but also a priority environmental pollutant because of its toxicity to higher organisms. In our previous studies, a gene cluster implicated to be involved in 4-cresol catabolism, creCDEFGHIR, was identified in Corynebacterium glutamicum and partially characterized in vivo. In this work, we report on the discovery of a novel 4-cresol biodegradation pathway that employs phosphorylated intermediates. This unique pathway initiates with the phosphorylation of the hydroxyl group of 4-cresol, which is catalyzed by a novel 4-methylbenzyl phosphate synthase, CreHI. Next, a unique class I P450 system, CreJEF, specifically recognizes phosphorylated intermediates and successively oxidizes the aromatic methyl group into carboxylic acid functionality via alcohol and aldehyde intermediates. Moreover, CreD (phosphohydrolase), CreC (alcohol dehydrogenase), and CreG (aldehyde dehydrogenase) were also found to be required for efficient oxidative transformations in this pathway. Steady-state kinetic parameters (Km and kcat) for each catabolic step were determined, and these results suggest that kinetic controls serve a key role in directing the metabolic flux to the most energy effective route.

  18. Bioremediation of refinery wastewater using immobilised Burkholderia cepacia and Corynebacterium sp and their transconjugants

    Directory of Open Access Journals (Sweden)

    Abdullahi T. Ajao

    2013-07-01

    Full Text Available When oil spill occurs, it poses serious toxic hazards to all forms of life. Mixed culture of Burkholderia cepacia and Corynebacterium sp isolated from refinery sludge using selective enrichment technique was used for bioremediation of refinery wastewater in a laboratoryscale bioreactor. Physicochemical parameters of both raw and treated water were as determined and compared with Federal Environ - mental Protection Agency (FEPA-limit, Abuja, Nigeria to asses the efficiency of the bioremediation process. Each of the bacterium was screened for the presence of plasmid DNA and for the involvement or otherwise of plasmid in the bioremediation of wastewater. The immobilised cells showed percentage decrease in chemical oxygen demand (97%, biochemical oxygen demand (94%, phenol (98%, total petroleum hydrocarbon (79%, oil and grease (90% of the refinery waste water after 20 days of treatment while their transconjugants showed the multiplicative effect by achieving the same percentage after 10 days of treatment. Therefore, the findings revealed that bioaugmentation of wastewater using transmissible catabolic plasmid will enhance efficiency of the bioremediation by spreading the plasmid among indigenous microbial community either through horizontal gene transfer or transformation.

  19. Detoxification of furfural in Corynebacterium glutamicum under aerobic and anaerobic conditions.

    Science.gov (United States)

    Tsuge, Yota; Hori, Yoshimi; Kudou, Motonori; Ishii, Jun; Hasunuma, Tomohisa; Kondo, Akihiko

    2014-10-01

    The toxic fermentation inhibitors in lignocellulosic hydrolysates raise serious problems for the microbial production of fuels and chemicals. Furfural is considered to be one of the most toxic compounds among these inhibitors. Here, we describe the detoxification of furfural in Corynebacterium glutamicum ATCC13032 under both aerobic and anaerobic conditions. Under aerobic culture conditions, furfuryl alcohol and 2-furoic acid were produced as detoxification products of furfural. The ratio of the products varied depending on the initial furfural concentration. Neither furfuryl alcohol nor 2-furoic acid showed any toxic effect on cell growth, and both compounds were determined to be the end products of furfural degradation. Interestingly, unlike under aerobic conditions, most of the furfural was converted to furfuryl alcohol under anaerobic conditions, without affecting the glucose consumption rate. Both the NADH/NAD(+) and NADPH/NADP(+) ratio decreased in the accordance with furfural concentration under both aerobic and anaerobic conditions. These results indicate the presence of a single or multiple endogenous enzymes with broad and high affinity for furfural and co-factors in C. glutamicum ATCC13032.

  20. Productivity of cyclohexanone oxidation of the recombinant Corynebacterium glutamicum expressing chnB of Acinetobacter calcoaceticus.

    Science.gov (United States)

    Doo, Eun-Hee; Lee, Won-Heong; Seo, Hyo-Seel; Seo, Jin-Ho; Park, Jin-Byung

    2009-06-15

    The biocatalytic efficiency of recombinant Corynebacterium glutamicum expressing the chnB gene encoding cyclohexanone monooxygenase (CHMO) of Acinetobacter calcoaceticus NCIMB 9871 was investigated. Optimization of an expression system and induction conditions enabled the recombinant biocatalyst to produce CHMO to a specific activity of ca. 0.5 U mg(-1) protein. Tight control of feeding of an energy source (i.e., glucose) and dissolved oxygen tension during fed-batch culture-based biotransformation allowed the cells to produce epsilon-caprolactone to a concentration of 16.0 g l(-1). The specific and volumetric productivity for cyclohexanone oxidation were 0.12 g g drycells(-1)h(-1) (17.5 U g(-1) of dry cells) and 2.3 g l(-1)h(-1) (330 U l(-1)), respectively. These values correspond to over 5.4- and 2.7-fold of recombinant Escherichia coli expressing the same gene under similar reaction conditions. It could be concluded that the recombinant C. glutamicum is a promising biocatalyst for Baeyer-Villiger oxidations.

  1. Effect of cysteine on methionine production by a regulatory mutant of Corynebacterium lilium

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Dharmendra; Subramanian, Kartik; Bisaria, Virendra S.; Sreekrishnan, T.R.; Gomes, James [Indian Inst. of Technology, Dept. of Biochemical Engineering and Biotechnology, New Delhi (India)

    2005-02-01

    The production of methionine by submerged fermentation using a mutant strain of Corynebacterium lilium was studied to determine suitable conditions for obtaining high productivity. The mutant strain resistant to the methionine analogues ethionine, norleucine, methionine sulfoxide and methionine methylsulfonium chloride produced 2.34 g l{sup -1} of methionine in minimal medium containing glucose as carbon source. The effect of cysteine on methionine production in a 15 l bioreactor was studied by supplementing cysteine intermittently during the course of fermentation. The addition of cysteine (0.75 g l{sup -1} h{sup -1}) every 2 h to the production medium increased the production of methionine to 3.39 g l{sup -1}. A metabolic flux analysis showed that during cysteine supplementation the ATP consumption reduced by 20%. It also showed that the increase in flux from phosphoenol pyruvate to oxaloacetate leads to higher methionine production. Results indicate that controlling the respiratory quotient close to 0.75 will produce the highest amount of methionine and that regulatory mutants also resistant to analogues of cysteine would be better methionine over producers. (Author)

  2. Dihydroxyacetone production in an engineered Escherichia coli through expression of Corynebacterium glutamicum dihydroxyacetone phosphate dephosphorylase.

    Science.gov (United States)

    Jain, Vishist Kumar; Tear, Crystal Jing Ying; Lim, Chan Yuen

    2016-05-01

    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 production. The accumulated DHAP was then converted to DHA through the heterologous expression of Corynebacterium glutamicum DHAP dephosphorylase (cghdpA) gene. To conserve DHAP exclusively for DHA production we removed methylglyoxal synthase (mgsA) gene in the ΔtpiA strain. This drastically improved DHA production from 0.83g/l (0.06g DHA/g glucose) in the ΔtpiA strain bearing cghdpA to 5.84g/l (0.41g DHA/g glucose) in the ΔtpiAΔmgsA double mutant containing the same gene. To limit the conversion of intracellular DHA to glycerol, glycerol dehydrogenase (gldA) gene was further knocked out resulting in a ΔtpiAΔmgsAΔgldA triple mutant. This triple mutant expressing the cghdpA gene produced 6.60g/l of DHA at 87% of the maximum theoretical yield. In summary, we demonstrated an efficient system for DHA production in genetically engineered E. coli strain. PMID:26992791

  3. Three-stage fermentation and kinetic modeling of bioflocculant by Corynebacterium glutamicum

    Institute of Scientific and Technical Information of China (English)

    Liang Shen; Zhongtao An; Qingbiao Li; Chuanyi Yao; Yajuan Peng; Yuanpeng Wang; Ruihua Lai; Xu Deng; Ning He

    2015-01-01

    Fermentation of bioflocculant with Corynebacterium glutamicum was studied by way of kinetic modeling. Lorentzian modified Logistic model, time-corrected Luedeking–Piret and Luedeking–Piret type models were pro-posed and applied to describe the cell growth, bioflocculant synthesis and consumption of substrates, with the correlation of initial biomass concentration and initial glucose concentration, respectively. The results showed that these models could well characterize the batch culture process of C. glutamicum at various initial glucose con-centrations from 10.0 to 17.5 g·L−1. The initial biomass concentration could shorten the lag time of cel growth, while the maximum biomass concentration was achieved only at the optimal initial glucose concentration of 16.22 g·L−1. A novel three-stage fed-batch strategy for bioflocculant production was developed based on the model prediction, in which the lag phase, quick biomass growth and bioflocculant production stages were sequentially proceeded with the adjustment of glucose concentration and dissolved oxygen. Biomass of 2.23 g·L−1 was obtained and bioflocculant concentration was enhanced to 176.32 mg·L−1, 18.62% and 403.63%higher than those in the batch process, respectively, indicating an efficient fed-batch culture strategy for bioflocculant production.

  4. Succinic acid production from corn cob hydrolysates by genetically engineered Corynebacterium glutamicum.

    Science.gov (United States)

    Wang, Chen; Zhang, Hengli; Cai, Heng; Zhou, Zhihui; Chen, Yilu; Chen, Yali; Ouyang, Pingkai

    2014-01-01

    Corynebacterium glutamicum wild type lacks the ability to utilize the xylose fractions of lignocellulosic hydrolysates. In the present work, we constructed a xylose metabolic pathway in C. glutamicum by heterologous expression of the xylA and xylB genes coming from Escherichia coli. Dilute-acid hydrolysates of corn cobs containing xylose and glucose were used as a substrate for succinic acid production by recombinant C. glutamicum NC-2. The results indicated that the available activated charcoal pretreatment in dilute-acid hydrolysates of corn cobs could be able to overcome the inhibitory effect in succinic acid production. Succinic acid was shown to be efficiently produced from corn cob hydrolysates (55 g l(-1) xylose and 4 g l(-1) glucose) under oxygen deprivation with addition of sodium carbonate. Succinic acid concentration reached 40.8 g l(-1) with a yield of 0.69 g g(-1) total sugars within 48 h. It was the first report of succinic acid production from corn cob hydrolysates by metabolically engineered C. glutamicum. This study suggested that dilute-acid hydrolysates of corn cobs may be an alternative substrate for the efficient production of succinic acid by C. glutamicum. PMID:24078255

  5. PARÁMETROS FISICOQUÍMICOS PARA LA SÍNTESIS DE ÁCIDO LÁCTICO O ETANOL DE LA BACTERIA (Corynebacterium glutamicum) Physico-Chemical Parameter for Production of Lactic Acid or Ethanol of (Corynebacterium glutamicum) Bacteria

    OpenAIRE

    ANGÉLICA CASTELLANOS; LINA MARCELA GARCIA; MYRIAM ASTUDILLO; JORGE ENRIQUE LÓPEZ GALÁN; LUZ MARINA FLOREZ PARDO

    2011-01-01

    El interés por obtener productos para la industria de biocombustibles a partir de desechos agrícolas, conduce a la búsqueda de nuevos sistemas biotecnológicos resistentes y costo-efectivos. Corynebacterium glutamicum, es un microorganismo usado para producir amino-ácidos que crece en gran variedad de sustratos y es resistente durante la fermentación, a variaciones de pH, temperatura, presión osmótica y acumulación de alcohol, características que lo hacen candidato a ser mejorado para la sínte...

  6. Cystic Neutrophilic Granulomatous Mastitis: Further Characterization of a Distinctive Histopathologic Entity Not Always Demonstrably Attributable to Corynebacterium Infection.

    Science.gov (United States)

    D'Alfonso, Timothy M; Moo, Tracy-Ann; Arleo, Elizabeth K; Cheng, Esther; Antonio, Lilian B; Hoda, Syed A

    2015-10-01

    Granulomatous lobular mastitis (GLM) is an uncommon condition that typically occurs in parous, reproductive-aged women and can simulate malignancy on the basis of clinical and imaging features. A distinctive histologic pattern termed cystic neutrophilic granulomatous mastitis (CNGM) is seen in some cases of GLM and has been associated with Corynebacterium infection. We sought to further characterize the clinical, imaging, and histopathologic features of CNGM by studying 12 cases and attempted to establish the relationship of this disease with Corynebacterium infection. Patients were women ranging in age from 25 to 49 years (median: 34 y), and all presented with a palpable mass that was painful in half of the cases. In 2 of 9 cases, imaging was highly suspicious for malignancy (BI-RADS 5). CNGM was characterized by lobulocentric granulomas with mixed inflammation and clear vacuoles lined by neutrophils within granulomas. Gram-positive bacilli were identified in 5/12 cases. In 4 patients, the disease process worsened after the diagnostic core biopsy, with the development of a draining sinus in 2 cases. No growth of bacteria was seen in any microbial cultures. No bacterial DNA was identified by 16S rDNA polymerase chain reaction for 1 case that showed gram-positive bacilli on histology. Patients were treated with variable combinations of surgery, antibiotics, and steroids. The time to significant resolution of symptoms ranged from 2 weeks to 6 months. Similar to other forms of GLM, CNGM can mimic malignancy clinically and on imaging. When encountered in a needle core biopsy sample, recognition of the characteristic histologic pattern and its possible association with Corynebacterium infection can help guide treatment. PMID:26200100

  7. Global Expression Profiling and Physiological Characterization of Corynebacterium glutamicum Grown in the Presence of l-Valine

    OpenAIRE

    Lange, C.; Rittmann, D.; Wendisch, V. F.; Bott, M.; Sahm, H

    2003-01-01

    Addition of l-valine (50 to 200 mM) to glucose minimal medium had no effect on the growth of wild-type Corynebacterium glutamicum ATCC 13032 but inhibited the growth of the derived valine production strain VAL1 [13032 ΔilvA ΔpanBC(pJC1ilvBNCD)] in a concentration-dependent manner. In order to explore this strain-specific valine effect, genomewide expression profiling was performed using DNA microarrays, which showed that valine caused an increased ilvBN mRNA level in VAL1 but not in the wild ...

  8. Improvement of the Redox Balance Increases l-Valine Production by Corynebacterium glutamicum under Oxygen Deprivation Conditions

    OpenAIRE

    Hasegawa, Satoshi; Uematsu, Kimio; Natsuma, Yumi; Suda, Masako; Hiraga, Kazumi; Jojima, Toru; Inui, Masayuki; Yukawa, Hideaki

    2012-01-01

    Production of l-valine under oxygen deprivation conditions by Corynebacterium glutamicum lacking the lactate dehydrogenase gene ldhA and overexpressing the l-valine biosynthesis genes ilvBNCDE was repressed. This was attributed to imbalanced cofactor production and consumption in the overall l-valine synthesis pathway: two moles of NADH was generated and two moles of NADPH was consumed per mole of l-valine produced from one mole of glucose. In order to solve this cofactor imbalance, the coenz...

  9. Global expression profiling and physiological characterization of Corynebacterium glutamicum grown in the presence of L-valine

    OpenAIRE

    Lange, C.; Rittmann, D.; Wendisch, V. F.; Bott, M.; Sahm, H

    2003-01-01

    Addition of L-valine (50 to 200 mM) to glucose minimal medium had no effect on the growth of wild-type Corynebacterium glutamicum ATCC 13032 but inhibited the growth of the derived valine production strain VAL1 [13032 DeltailvA DeltapanBC(pJCilvBNCD)] in a concentration-dependent manner. In order to explore this strain-specific valine effect, genomewide expression profiling was performed using DNA microarrays, which showed that valine caused an increased ilvBN mRNA level in VAL1 but not in th...

  10. Enhanced Fc receptor expression by a sub-population of murine intra-tumour macrophages following intravenous Corynebacterium parvum therapy.

    OpenAIRE

    Moore, K; McBride, W.H.

    1983-01-01

    Intravenous injection of Corynebacterium parvum (C. parvum) 4 days after s.c. inoculation of 5 X 10(5) cells derived from the immunogenic fibrosarcoma FSA/R induced tumour growth inhibition over a period of 21 days in syngeneic C3H/Buf mice. This was not accompanied by a change in the proportions of host cells within the tumour, but the activation state of tumour-infiltrating macrophages was increased following C. parvum therapy. Two macrophage subpopulations were identified in FSA/R tumours ...

  11. Utilization of fermentation waste (Corynebacterium glutamicum) for biosorption of Reactive Black 5 from aqueous solution

    International Nuclear Information System (INIS)

    A fermentation waste, Corynebacterium glutamicum, was successfully employed as a biosorbent for Reactive Black 5 (RB5) from aqueous solution. This paper initially studied the effect of pretreatment on the biosorption capacity of C. glutamicum toward RB5, using several chemical agents, such as HCl, H2SO4, HNO3, NaOH, Na2CO3, CaCl2 and NaCl. Among these reagents, 0.1 M HNO3 gave the maximum enhancement of the RB5 uptake, exhibiting 195 mg/g at pH 1 with an initial RB5 concentration of 500 mg/l. The solution pH and temperature were found to affect the biosorption capacity, and the biosorption isotherms derived at different pHs and temperatures revealed that a low pH (pH 1) and high temperature (35 deg. C) favored biosorption. The biosorption isotherm was well represented using three-parameter models (Redlich-Peterson and Sips) compared to two-parameter models (Langmuir and Freundlich models). As a result, high correlation coefficients and low average percentage error values were observed for three-parameter models. A maximum RB5 uptake of 419 mg/g was obtained at pH 1 and a temperature of 35 deg. C, according to the Langmuir model. The kinetics of the biosorption process with different initial concentrations (500-2000 mg/l) was also monitored, and the data were analyzed using pseudo-first and pseudo-second order models, with the latter describing the data well. Various thermodynamic parameters, such as ΔGo, ΔHo and ΔSo, were calculated, indicating that the present system was a spontaneous and endothermic process. The use of a 0.1 M NaOH solution successfully desorbed almost all the dye molecules from dye-loaded C. glutamicum biomass at different solid-to-liquid ratios examined

  12. Implantation of Corynebacterium pseudodiphtheriticum for elimination of Staphylococcus aureus from the nasal cavity in volunteers

    Science.gov (United States)

    Viacheslav, Ilyin; Kiryukhina, Nataliya

    Nasal carriage of Staphylococcus aureus is a well-documented risk factor of infection and inflammation of the skin, soft tissues and bacteremia. It is also known that most often etiology of these disorders is associated with autoinfection. The present-day methods of opportunistic pathogens eradication from the nasal cavity are based principally on the use of antiseptic and antibacterial agents. For instance, a local antibiotic mupirocin in the form of nasal ointment is considered to be the gold standard for the treatment of S. aureus carriage. The literature describes investigations showing how mupirocin can strengthen antibiotic resistance in S. aureus strains, including those with methicillin resistance (MRSA). It is also common knowledge that recolonization of the nasal mucous membrane takes place within several months after mupirocin treatment. This circumstance dictates the necessity to look for alternative ways of preventing the S. aureus carriage and methods of elimination. One of the methods of nasal S. aureus elimination is implantation of nonpathogenic microorganisms which will extrude opportunistic pathogens without impinging the symbiotic microbiota. Effectiveness of saline suspension of Corynebacterium pseudodiphtheriticum containing spray was assessed in a several chamber experiments with simulation of some spaceflight factors (dry immersion, isolation). Various schemes of application of preparations were applied. In all cases of corynebacteria application the strong inhibiting effect against S. aureus was detected. This fact opens a prospect of using nonpathogenic corynebacteria as a nasal probiotic. Administration of the nasal corynebacteria spray possibly prevented cross-infection by MRSA and appearance of staphylococcal infection. Further pre-clinical and clinical study of this bacterial therapy method is under development.

  13. Biofilm formation and fibrinogen and fibronectin binding activities by Corynebacterium pseudodiphtheriticum invasive strains.

    Science.gov (United States)

    Souza, Monica Cristina; dos Santos, Louisy Sanches; Sousa, Leonardo Paiva; Faria, Yuri Vieira; Ramos, Juliana Nunes; Sabbadini, Priscila Soares; da Santos, Cíntia Silva; Nagao, Prescilla Emy; Vieira, Verônica Viana; Gomes, Débora Leandro Rama; Hirata Júnior, Raphael; Mattos-Guaraldi, Ana Luiza

    2015-06-01

    Biofilm-related infections are considered a major cause of morbidity and mortality in hospital environments. Biofilms allow microorganisms to exchange genetic material and to become persistent colonizers and/or multiresistant to antibiotics. Corynebacterium pseudodiphtheriticum (CPS), a commensal bacterium that colonizes skin and mucosal sites has become progressively multiresistant and responsible for severe nosocomial infections. However, virulence factors of this emergent pathogen remain unclear. Herein, we report the adhesive properties and biofilm formation on hydrophilic (glass) and hydrophobic (plastic) abiotic surfaces by CPS strains isolated from patients with localized (ATCC10700/Pharyngitis) and systemic (HHC1507/Bacteremia) infections. Adherence to polystyrene attributed to hydrophobic interactions between bacterial cells and this negatively charged surface indicated the involvement of cell surface hydrophobicity in the initial stage of biofilm formation. Attached microorganisms multiplied and formed microcolonies that accumulated as multilayered cell clusters, a step that involved intercellular adhesion and synthesis of extracellular matrix molecules. Further growth led to the formation of dense bacterial aggregates embedded in the exopolymeric matrix surrounded by voids, typical of mature biofilms. Data also showed CPS recognizing human fibrinogen (Fbg) and fibronectin (Fn) and involvement of these sera components in formation of "conditioning films". These findings suggested that biofilm formation may be associated with the expression of different adhesins. CPS may form biofilms in vivo possibly by an adherent biofilm mode of growth in vitro currently demonstrated on hydrophilic and hydrophobic abiotic surfaces. The affinity to Fbg and Fn and the biofilm-forming ability may contribute to the establishment and dissemination of infection caused by CPS.

  14. Engineering of Corynebacterium glutamicum for high-yield L-valine production under oxygen deprivation conditions.

    Science.gov (United States)

    Hasegawa, Satoshi; Suda, Masako; Uematsu, Kimio; Natsuma, Yumi; Hiraga, Kazumi; Jojima, Toru; Inui, Masayuki; Yukawa, Hideaki

    2013-02-01

    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 ldhA. Nonetheless, a few other by-products, particularly succinate, were still produced and acted to suppress the L-valine yield. Eliminating these by-products therefore was deemed key to improving theL-valine yield. By additionally disrupting the phosphoenolpyruvate carboxylase gene ppc, succinate production was effectively suppressed, but both glucose consumption and L-valine production dropped considerably due to the severely elevated intracellular NADH/NAD(+) ratio. In contrast, this perturbed intracellular redox state was more than compensated for by deletion of three genes associated with NADH-producing acetate synthesis and overexpression of five glycolytic genes, including gapA, encoding NADH-inhibited glyceraldehyde-3-phosphate dehydrogenase. Inserting feedback-resistant mutant acetohydroxy acid synthase and NAD-preferring mutant AHAIR in the chromosome resulted in higher L-valine yield and productivity. Deleting the alanine transaminase gene avtA suppressed alanine production. The resultant strain produced 1,280 mM L-valine at a yield of 88% mol mol of glucose(-1) after 24 h under oxygen deprivation, a vastly improved yield over our previous best.

  15. Targets of nitric oxide in a mouse model of liver inflammation by Corynebacterium parvum.

    Science.gov (United States)

    Chamulitrat, W; Jordan, S J; Mason, R P; Litton, A L; Wilson, J G; Wood, E R; Wolberg, G; Molina y Vedia, L

    1995-01-10

    Treatment of mice with Corynebacterium parvum induces chronic inflammation. This treatment followed by an injection of lipopolysaccharide (LPS) produces hepatic necrosis and death. We examined liver tissue by using electron paramagnetic resonance (EPR) spectroscopy and found that, in addition to the previously reported nonheme nitrosyl complexes, heme nitrosyl complexes were also formed. Hemoglobin nitrosyl complexes measured in the whole blood of mice treated with C. parvum were not increased after additional LPS treatment. However, this treatment significantly increased the heme nitrosyl complexes in the liver, whereas the nonheme nitrosyl complex concentration was unaffected. EPR signals from whole blood and liver tissues from mice treated with C. parvum and C. parvum + LPS were inhibited by prolonged treatment with NG-monomethyl-L-arginine (L-NMA). Nitric oxide (.NO) is known to bind to cytochrome P450 heme, and we consistently found a suppression of EPR signals attributable to ferric low-spin cytochrome P450/P420 peaks in the livers of mice treated with C. parvum and C. parvum + LPS. By performing analyses of EPR spectra obtained from hepatocytes exposed to .NO, we were able to unambiguously identify EPR signals attributable to cytochrome P420 and nonheme nitrosyl complexes in the livers of both treatments. Deconvolution of the composite in vivo EPR spectra indicated that hemoglobin nitrosyl complexes contributed weakly in the C. parvum livers, but threefold more in the C. parvum + LPS livers, suggesting that hemorrhage may have occurred. Experiments with L-NMA treatment revealed that this additional .NO production did not correlate with hepatic necrosis and onset of death. Immunoprecipitation of liver cytosols from C. parvum- and (C. parvum + LPS)-treated mice using an antibody against mouse inducible nitric oxide synthase showed that this enzyme was indeed present in the cytosolic fractions and was absent in those from control livers. Our novel detection of

  16. Pushing product formation to its limit: metabolic engineering of Corynebacterium glutamicum for L-leucine overproduction.

    Science.gov (United States)

    Vogt, Michael; Haas, Sabine; Klaffl, Simon; Polen, Tino; Eggeling, Lothar; van Ooyen, Jan; Bott, Michael

    2014-03-01

    Using metabolic engineering, an efficient L-leucine production strain of Corynebacterium glutamicum was developed. In the wild type of C. glutamicum, the leuA-encoded 2-isopropylmalate synthase (IPMS) is inhibited by low L-leucine concentrations with a K(i) of 0.4 mM. We identified a feedback-resistant IMPS variant, which carries two amino acid exchanges (R529H, G532D). The corresponding leuA(fbr) gene devoid of the attenuator region and under control of a strong promoter was integrated in one, two or three copies into the genome and combined with additional genomic modifications aimed at increasing L-leucine production. These modifications involved (i) deletion of the gene encoding the repressor LtbR to increase expression of leuBCD, (ii) deletion of the gene encoding the transcriptional regulator IolR to increase glucose uptake, (iii) reduction of citrate synthase activity to increase precursor supply, and (iv) introduction of a gene encoding a feedback-resistant acetohydroxyacid synthase. The production performance of the resulting strains was characterized in bioreactor cultivations. Under fed-batch conditions, the best producer strain accumulated L-leucine to levels exceeding the solubility limit of about 24 g/l. The molar product yield was 0.30 mol L-leucine per mol glucose and the volumetric productivity was 4.3 mmol l⁻¹ h⁻¹. These values were obtained in a defined minimal medium with a prototrophic and plasmid-free strain, making this process highly interesting for industrial application. PMID:24333966

  17. Production of 2-ketoisocaproate with Corynebacterium glutamicum strains devoid of plasmids and heterologous genes.

    Science.gov (United States)

    Vogt, Michael; Haas, Sabine; Polen, Tino; van Ooyen, Jan; Bott, Michael

    2015-03-01

    2-Ketoisocaproate (KIC), the last intermediate in l-leucine biosynthesis, has various medical and industrial applications. After deletion of the ilvE gene for transaminase B in l-leucine production strains of Corynebacterium glutamicum, KIC became the major product, however, the strains were auxotrophic for l-isoleucine. To avoid auxotrophy, reduction of IlvE activity by exchanging the ATG start codon of ilvE by GTG was tested instead of an ilvE deletion. The resulting strains were indeed able to grow in glucose minimal medium without amino acid supplementation, but at the cost of lowered growth rates and KIC production parameters. The best production performance was obtained with strain MV-KICF1, which carried besides the ilvE start codon exchange three copies of a gene for a feedback-resistant 2-isopropylmalate synthase, one copy of a gene for a feedback-resistant acetohydroxyacid synthase and deletions of ltbR and iolR encoding transcriptional regulators. In the presence of 1 mM l-isoleucine, MV-KICF1 accumulated 47 mM KIC (6.1 g l(-1)) with a yield of 0.20 mol/mol glucose and a volumetric productivity of 1.41 mmol KIC l(-1)  h(-1). Since MV-KICF1 is plasmid free and lacks heterologous genes, it is an interesting strain for industrial application and as platform for the production of KIC-derived compounds, such as 3-methyl-1-butanol. PMID:25488800

  18. Crystal structures of halohydrin hydrogen-halide-lyases from Corynebacterium sp. N-1074.

    Science.gov (United States)

    Watanabe, Fumiaki; Yu, Fujio; Ohtaki, Akashi; Yamanaka, Yasuaki; Noguchi, Keiichi; Yohda, Masafumi; Odaka, Masafumi

    2015-12-01

    Halohydrin hydrogen-halide-lyase (H-Lyase) is a bacterial enzyme that is involved in the degradation of halohydrins. This enzyme catalyzes the intramolecular nucleophilic displacement of a halogen by a vicinal hydroxyl group in halohydrins to produce the corresponding epoxides. The epoxide products are subsequently hydrolyzed by an epoxide hydrolase, yielding the corresponding 1, 2-diol. Until now, six different H-Lyases have been studied. These H-Lyases are grouped into three subtypes (A, B, and C) based on amino acid sequence similarities and exhibit different enantioselectivity. Corynebacterium sp. strain N-1074 has two different isozymes of H-Lyase, HheA (A-type) and HheB (B-type). We have determined their crystal structures to elucidate the differences in enantioselectivity among them. All three groups share a similar structure, including catalytic sites. The lack of enantioselectivity of HheA seems to be due to the relatively wide size of the substrate tunnel compared to that of other H-Lyases. Among the B-type H-Lyases, HheB shows relatively high enantioselectivity compared to that of HheBGP1 . This difference seems to be due to amino acid replacements at the active site tunnel. The binding mode of 1, 3-dicyano-2-propanol at the catalytic site in the crystal structure of the HheB-DiCN complex suggests that the product should be (R)-epichlorohydrin, which agrees with the enantioselectivity of HheB. Comparison with the structure of HheC provides a clue for the difference in their enantioselectivity.

  19. Selection and Characterization of a Lysine Yielding Mutant of Corynebacterium glutamicum - a Soil Isolate from Pakistan

    Directory of Open Access Journals (Sweden)

    Habib-ur-Rehman§٭, Abdul Hameed and Safia Ahmed

    2012-01-01

    Full Text Available L-lysine is the second limiting amino acid for poultry and supplemented in broiler feed for optimal performance. Lysine can be produced by inducing mutation in glutamate producing bacteria. The study was conducted to enhance lysine production from a local strain of Corynebacterium glutamicum. The bacterium was mutated by exposure to UV. Mutants resistant to s-2-aminoethyle L-cystein (AEC and showing auxotrophy for L-homoserine were screened for lysine production qualitatively and quantitatively. A mutant showing highest production of lysine (8.2 mg/mL was selected for optimization of physical and nutritional parameters for maximum production of lysine in shake flask. An initial pH 7.6, 30˚C temperature, 300 rpm and 60 h incubation time were the optimized values of physical requirements. Cane molasses and corn starch hydrolysate were required at 15% (w/v in the fermentation media which provided around 9% total sugars to produce maximum lysine (17 to 18 mg/mL. When amonium sulphate was used at 3.5% (w/v level in molasses or corn starch hydrolysate based fermentation media, production of lysine slightly increased above 18 mg/mL. It is concluded that industrial by products like cane molasses, corn steep liquor, and corn starch hydrolysate can be used as carbon and organic nitrogen sources in fermentation medium for scale up process of lysine production and this lysine enriched broth may be used in broiler feed later. However, more potent lysine producing mutant and additional in vivo trials would be required to commercialize this product.

  20. Metabolic engineering of Corynebacterium glutamicum to produce GDP-L-fucose from glucose and mannose.

    Science.gov (United States)

    Chin, Young-Wook; Park, Jin-Byung; Park, Yong-Cheol; Kim, Kyoung Heon; Seo, Jin-Ho

    2013-06-01

    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-fucose from GDP-D-mannose. Coexpression of the genes allowed the recombinant C. glutamicum cells to produce GDP-L-fucose in a minimal medium containing glucose and mannose as carbon sources. The specific product formation rate was much higher during growth on mannose than on glucose. In addition, the specific product formation rate was further increased by coexpressing the endogenous phosphomanno-mutase gene (manB) and GTP-mannose-1-phosphate guanylyl-transferase gene (manC), which are involved in the conversion of mannose-6-phosphate into GDP-D-mannose. However, the overexpression of manA encoding mannose-6-phosphate isomerase, catalyzing interconversion of mannose-6-phosphate and fructose-6-phosphate showed a negative effect on formation of the target product. Overall, coexpression of gmd, wcaG, manB and manC in C. glutamicum enabled production of GDP-L-fucose at the specific rate of 0.11 mg g cell(-1) h(-1). The specific GDP-L-fucose content reached 5.5 mg g cell(-1), which is a 2.4-fold higher than that of the recombinant E. coli overexpressing gmd, wcaG, manB and manC under comparable conditions. Well-established metabolic engineering tools may permit optimization of the carbon and cofactor metabolisms of C. glutamicum to further improve their production capacity.

  1. Proteome scale comparative modeling for conserved drug and vaccine targets identification in Corynebacterium pseudotuberculosis.

    Science.gov (United States)

    Hassan, Syed Shah; Tiwari, Sandeep; Guimarães, Luís Carlos; Jamal, Syed Babar; Folador, Edson; Sharma, Neha Barve; de Castro Soares, Siomar; Almeida, Síntia; Ali, Amjad; Islam, Arshad; Póvoa, Fabiana Dias; de Abreu, Vinicius Augusto Carvalho; Jain, Neha; Bhattacharya, Antaripa; Juneja, Lucky; Miyoshi, Anderson; Silva, Artur; Barh, Debmalya; Turjanski, Adrian Gustavo; Azevedo, Vasco; Ferreira, Rafaela Salgado

    2014-01-01

    Corynebacterium pseudotuberculosis (Cp) is a pathogenic bacterium that causes caseous lymphadenitis (CLA), ulcerative lymphangitis, mastitis, and edematous to a broad spectrum of hosts, including ruminants, thereby threatening economic and dairy industries worldwide. Currently there is no effective drug or vaccine available against Cp. To identify new targets, we adopted a novel integrative strategy, which began with the prediction of the modelome (tridimensional protein structures for the proteome of an organism, generated through comparative modeling) for 15 previously sequenced C. pseudotuberculosis strains. This pan-modelomics approach identified a set of 331 conserved proteins having 95-100% intra-species sequence similarity. Next, we combined subtractive proteomics and modelomics to reveal a set of 10 Cp proteins, which may be essential for the bacteria. Of these, 4 proteins (tcsR, mtrA, nrdI, and ispH) were essential and non-host homologs (considering man, horse, cow and sheep as hosts) and satisfied all criteria of being putative targets. Additionally, we subjected these 4 proteins to virtual screening of a drug-like compound library. In all cases, molecules predicted to form favorable interactions and which showed high complementarity to the target were found among the top ranking compounds. The remaining 6 essential proteins (adk, gapA, glyA, fumC, gnd, and aspA) have homologs in the host proteomes. Their active site cavities were compared to the respective cavities in host proteins. We propose that some of these proteins can be selectively targeted using structure-based drug design approaches (SBDD). Our results facilitate the selection of C. pseudotuberculosis putative proteins for developing broad-spectrum novel drugs and vaccines. A few of the targets identified here have been validated in other microorganisms, suggesting that our modelome strategy is effective and can also be applicable to other pathogens. PMID:25573232

  2. Evaluation and characterisation of A and B fragments of Corynebacterium diphtheriae toxin towards recombinant diphtheria vaccine

    Directory of Open Access Journals (Sweden)

    S Abulmagd

    2013-01-01

    Full Text Available Background: Diphtheria is a highly communicable disease caused by toxin-producing strains of Corynebacterium diphtheriae. Objectives: To evaluate the efficacy of A and B subunits of diphtheria toxin (DT-A, DT-B as potential vaccines against C. diphtheriae. A culture of C. diphtheriae (strain PW 8 was grown on Loeffler plates while Lingood medium was used for production of diphtheria toxin (DT. Materials and Methods: DT was purified and digested to obtain pure DT-A and DT-B and detoxified to obtain diphtheria toxin. Four groups of mice were immunised with different antigens (Ag of C. diphtheriae. Results: The antibody (Ab titres were significantly increased with immunised groups subsequent to three injections. On the other hand, Ab titres were estimated after the three immunisations and the levels of different Ab isotypes were comparatively measured. The levels of various isotypes immune responses showed variation between immunised groups where the IgG subclasses were significantly increased mainly with DPT immunised group. The IgM and IgA were significantly increased with DT-A more than others. Additionally, the evaluation of the cellular immune responses demonstrated that spleen cells from DPT and DT-A groups gave highly significant proliferative response with production of high levels of IL-2 and IFN-γ (Th1/Th2. Separation and purification of DT gene were performed using polymerase chain reaction (PCR and sub-cloned in pGEM-T vector, for further studying of recombinant vaccine. Conclusion: Our results showed the possibility to prepare a potent recombinant vaccine containing whole DT gene or DT-A against C. diphtheriae or could be used in treatment of cancer as it give high levels of IL-2 and IFN-γ.

  3. Engineering a Lysine-ON Riboswitch for Metabolic Control of Lysine Production in Corynebacterium glutamicum.

    Science.gov (United States)

    Zhou, Li-Bang; Zeng, An-Ping

    2015-12-18

    Riboswitches are natural RNA elements that regulate gene expression by binding a ligand. Here, we demonstrate the possibility of altering a natural lysine-OFF riboswitch from Eschericia coli (ECRS) to a synthetic lysine-ON riboswitch and using it for metabolic control. To this end, a lysine-ON riboswitch library was constructed using tetA-based dual genetic selection. After screening the library, the functionality of the selected lysine-ON riboswitches was examined using a report gene, lacZ. Selected lysine-ON riboswitches were introduced into the lysE gene (encoding a lysine transport protein) of Corynebacterium glutamicum and used to achieve dynamic control of lysine transport in a recombinant lysine-producing strain, C. glutamicum LPECRS, which bears a deregulated aspartokinase and a lysine-OFF riboswitch for dynamic control of the enzyme citrate synthase. Batch fermentation results of the strains showed that the C. glutamicum LPECRS strain with an additional lysine-ON riboswitch for the control of lysE achieved a 21% increase in the yield of lysine compared to that of the C. glutamicum LPECRS strain and even a 89% increase in yield compared to that of the strain with deregulated aspartokinase. This work provides a useful approach to generate lysine-ON riboswitches for C. glutamicum metabolic engineering and demonstrates for the first time a synergetic effect of lysine-ON and -OFF riboswitches for improving lysine production in this industrially important microorganism. The approach can be used to dynamically control other genes and can be applied to other microorganisms. PMID:26300047

  4. Engineering of Corynebacterium glutamicum for xylitol production from lignocellulosic pentose sugars.

    Science.gov (United States)

    Dhar, Kiran S; Wendisch, Volker F; Nampoothiri, Kesavan Madhavan

    2016-07-20

    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-psicose 3 epimerase (dpe) and l-xylulose reductase (lxr) genes from Escherichia coli, Agrobacterium tumefaciens and Mycobacterium smegmatis, respectively. Expression of xr and the synthetic araA-dpe-lxr operon under the control of IPTG-inducible Ptac promoter enabled production of xylitol from both xylose and arabinose in the mineral (CGXII) medium with glucose as carbon source. Additional expression of a pentose transporter (araTF) gene enhanced xylitol production by about four-fold compared to the parent strain. The constructed strain Cg-ax3 produced 6.7±0.4g/L of xylitol in batch fermentations and 31±0.5g/L of xylitol in fed-batch fermentations with a specific productivity of 0.28±0.05g/g cdw/h. The strain Cg-ax3 was also validated for xylitol production from pentose rich, acid pre-treated liquor of sorghum stover (SAPL) and the results were comparable in both SAPL (27±0.3g/L) and mineral medium (31±0.5g/L). PMID:27184428

  5. Impact of different CO2/HCO3- levels on metabolism and regulation in Corynebacterium glutamicum.

    Science.gov (United States)

    Blombach, Bastian; Buchholz, Jens; Busche, Tobias; Kalinowski, Jörn; Takors, Ralf

    2013-12-01

    We investigated the growth kinetics and transcriptional responses of Corynebacterium glutamicum in environments with low (pCO2CO2/HCO3(-) levels compared to standard conditions. When cultivated at high CO2/HCO3(-)-levels, C. glutamicum showed increased (63%) biomass to substrate yields during the initial growth phase. Other kinetic parameters such as growth rate (μ), specific glucose consumption rate (qS), and selected enzymatic activities of anaplerotic reactions, the pentose phosphate pathway and the tricarboxylic acid cycle were similar to standard conditions. However, microarray hybridization disclosed a complex transcriptional response involving 117 differentially expressed genes. Among those, 60 genes were assigned to the complete DtxR/RipA regulon controlling iron homeostasis in C. glutamicum. Impaired growth of a ΔdtxR mutant at high CO2/HCO3(-) levels validated the relevance of this master regulator to cope with excessive CO2/HCO3(-) availability. At low CO2/HCO3(-) levels, C. glutamicum grew in a bi-level manner with three distinct growth phases. Differential analyses revealed approximately doubled activities of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase accompanied by the formation of L-alanine and L-valine during the lowest μ occurring in mid-phase of the cultivation. DNA microarray analysis revealed more than 100 differentially expressed genes in growth phase II compared to phase I including almost all thiamin pyrophosphate (TPP) biosynthesis genes, which were significantly up regulated. Concluding, we hypothesize that C. glutamicum counteracts the lack of CO2/HCO3(-) by triggering TPP biosynthesis for increasing the activities of TPP-dependent enzymes involved in CO2 formation.

  6. Effect of non-cell Corynebacterium Parvum on differentiation and maturation of bone marrow-derived dendritic cells.

    Science.gov (United States)

    Liu, Chenghu; Gao, Shangxian; Qu, Zhonghua; Guo, Chun; Wu, Ping; Shi, Yanping; Zhang, Lining

    2012-01-01

    Corynebacterium parvum (CP), with their potent anti-tumor activities, has been well documented. Non-cell Corynebacterium Parvum (NCPP) is a neotype of biological preparation, which based on manipulating CP with nanotechnology. The present study was designed to investigate the effect of NCPP/CP on bone marrow derived dendritic cells (BMDCs) in tumor-bearing mice, especially focused on the differentiation and maturation of these BMDCs. BM cells from tumor-bearing mice administrated with NCPP/CP were analyzed by flow cytometry, which exhibit enhanced numbers of DCs and macrophages. In the meanwhile, flow cytometry analysis showed mild but significant difference for CD80 expression on these LPS- treated BMDCs between NCPP/CP administrated mice and the control animals. Furthermore, antigen presenting assay for these LPS-treated BMDCs showed significant difference for cytolytic assay of CD8+T cells against B16 melanoma cells, which indicate that NCPP treatments have enhanced the cytolytic rates of CD8+T cells from 47.9%±2.3% to 54.2%±2.4%. The data suggest that NCPP/CP treatment can efficiently facilitate the generation of BMDCs in vivo and enhance the maturation of these BMDCs in vitro. PMID:22676053

  7. l-Valine Production during Growth of Pyruvate Dehydrogenase Complex- Deficient Corynebacterium glutamicum in the Presence of Ethanol or by Inactivation of the Transcriptional Regulator SugR▿

    OpenAIRE

    Blombach, Bastian; Arndt, Annette; Auchter, Marc; Eikmanns, Bernhard J.

    2008-01-01

    Pyruvate dehydrogenase complex-deficient strains of Corynebacterium glutamicum produce l-valine from glucose only after depletion of the acetate required for growth. Here we show that inactivation of the DeoR-type transcriptional regulator SugR or replacement of acetate by ethanol already in course of the growth phase results in efficient l-valine production.

  8. Characterization of Cg10062 from Corynebacterium glutamicum : Implications for the evolution of cis-3-chloroacrylic acid dehalogenase activity in the tautomerase superfamily

    NARCIS (Netherlands)

    Poelarends, Gerrit J.; Serrano, Hector; Person, Maria D.; Johnson, William H.; Whitman, Christian P.

    2008-01-01

    A 149-amino acid protein designated Cg10062 is encoded by a gene from Corynebacterium glutamicum. The physiological function of Cg10062 is unknown, and the gene encoding this protein has no obvious genomic context. Sequence analysis links Cg10062 to the cis-3-chloroacrylic acid dehalogenase (cis-Caa

  9. The role of lipids and salts in two-dimensional crystallization of the glycine-betaine transporter BetP from Corynebacterium glutamicum

    DEFF Research Database (Denmark)

    Tsai, Ching-Ju; Ejsing, Christer S.; Shevchenko, Andrej;

    2007-01-01

    The osmoregulated and chill-sensitive glycine-betaine transporter (BetP) from Corynebacterium glutamicum was reconstituted into lipids to form two-dimensional (2D) crystals. The sensitivity of BetP partly bases on its interaction with lipids. Here we demonstrate that lipids and salts influence...

  10. Multiplex polymerase chain reaction to identify and determine the toxigenicity of Corynebacterium spp with zoonotic potential and an overview of human and animal infections

    Directory of Open Access Journals (Sweden)

    Luciene de Fatima Costa Torres

    2013-05-01

    Full Text Available Corynebacterium diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis constitute a group of potentially toxigenic microorganisms that are related to different infectious processes in animal and human hosts. Currently, there is a lack of information on the prevalence of disease caused by these pathogens, which is partially due to a reduction in the frequency of routine laboratory testing. In this study, a multiplex polymerase chain reaction (mPCR assay that can simultaneously identify and determine the toxigenicity of these corynebacterial species with zoonotic potential was developed. This assay uses five primer pairs targeting the following genes: rpoB (Corynebacterium spp, 16S rRNA (C. ulcerans and C. pseudotuberculosis, pld (C. pseudotuberculosis, dtxR (C. diphtheriae and tox [diphtheria toxin (DT ]. In addition to describing this assay, we review the literature regarding the diseases caused by these pathogens. Of the 213 coryneform strains tested, the mPCR results for all toxigenic and non-toxigenic strains of C . diphtheriae, C. ulcerans and C. pseudotuberculosis were in 100% agreement with the results of standard biochemical tests and PCR-DT. As an alternative to conventional methods, due to its advantages of specificity and speed, the mPCR assay used in this study may successfully be applied for the diagnosis of human and/or animal diseases caused by potentially toxigenic corynebacterial species.

  11. Engineering Corynebacterium glutamicum for fast production of L-lysine and L-pipecolic acid.

    Science.gov (United States)

    Pérez-García, Fernando; Peters-Wendisch, Petra; Wendisch, Volker F

    2016-09-01

    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 for the absence of the PTS in Δhpr strains. Growth of PTS-positive strains with glucose was accelerated when the endogenous inositol permease IolT2 and glucokinase from B. subtilis were overproduced with balanced translation initiation rates using plasmid pEKEx3-IolTBest. When the genome-reduced C. glutamicum strain GRLys1 carrying additional in-frame deletions of sugR and ldhA to derepress glycolytic and PTS genes and to circumvent formation of L-lactate as by-product was transformed with this plasmid or with pVWEx1-IolTBest, 18 to 20 % higher volumetric productivities and 70 to 72 % higher specific productivities as compared to the parental strain resulted. The non-proteinogenic amino acid L-pipecolic acid (L-PA), a precursor of immunosuppressants, peptide antibiotics, or piperidine alkaloids, can be derived from L-lysine. To enable production of L-PA by the constructed L-lysine-producing strain, the L-lysine 6-dehydrogenase gene lysDH from Silicibacter pomeroyi and the endogenous pyrroline 5-carboxylate reductase gene proC were overexpressed as synthetic operon. This enabled C. glutamicum to produce L-PA with a yield of 0.09 ± 0.01 g g(-1) and a volumetric productivity of 0.04 ± 0.01 g L(-1) h(-1).To the best of our knowledge, this is the first fermentative process for the production of L-PA from glucose.

  12. Gene Expression Analysis of Corynebacterium glutamicum Subjected to Long-Term Lactic Acid Adaptation▿ ¶

    Science.gov (United States)

    Jakob, Kinga; Satorhelyi, Peter; Lange, Christian; Wendisch, Volker F.; Silakowski, Barbara; Scherer, Siegfried; Neuhaus, Klaus

    2007-01-01

    Corynebacteria form an important part of the red smear cheese microbial surface consortium. To gain a better understanding of molecular adaptation due to low pH induced by lactose fermentation, the global gene expression profile of Corynebacterium glutamicum adapted to pH 5.7 with lactic acid under continuous growth in a chemostat was characterized by DNA microarray analysis. Expression of a total of 116 genes was increased and that of 90 genes was decreased compared to pH 7.5 without lactic acid, representing 7% of the genes in the genome. The up-regulated genes encode mainly transcriptional regulators, proteins responsible for export, import, and metabolism, and several proteins of unknown function. As much as 45% of the up-regulated open reading frames code for hypothetical proteins. These results were validated using real-time reverse transcription-PCR. To characterize the functions of 38 up-regulated genes, 36 single-crossover disruption mutants were generated and analyzed for their lactic acid sensitivities. However, only a sigB knockout mutant showed a highly significant negative effect on growth at low pH, suggesting a function in organic-acid adaptation. A sigE mutant already displayed growth retardation at neutral pH but grew better at acidic pH than the sigB mutant. The lack of acid-sensitive phenotypes in 34 out of 36 disrupted genes suggests either a considerable redundancy in acid adaptation response or coincidental effects. Other up-regulated genes included genes for ion transporters and metabolic pathways, including carbohydrate and respiratory metabolism. The enhanced expression of the nrd (ribonucleotide reductase) operon and a DNA ATPase repair protein implies a cellular response to combat acid-induced DNA damage. Surprisingly, multiple iron uptake systems (totaling 15% of the genes induced ≥2-fold) were induced at low pH. This induction was shown to be coincidental and could be attributed to iron-sequestering effects in complex media at low p

  13. Metabolic responses to pyruvate kinase deletion in lysine producing Corynebacterium glutamicum

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    Wittmann Christoph

    2008-03-01

    Full Text Available Abstract Background Pyruvate kinase is an important element in flux control of the intermediate metabolism. It catalyzes the irreversible conversion of phosphoenolpyruvate into pyruvate and is under allosteric control. In Corynebacterium glutamicum, this enzyme was regarded as promising target for improved production of lysine, one of the major amino acids in animal nutrition. In pyruvate kinase deficient strains the required equimolar ratio of the two lysine precursors oxaloacetate and pyruvate can be achieved through concerted action of the phosphotransferase system (PTS and phosphoenolpyruvate carboxylase (PEPC, whereby a reduced amount of carbon may be lost as CO2 due to reduced flux into the tricarboxylic acid (TCA cycle. In previous studies, deletion of pyruvate kinase in lysine-producing C. glutamicum, however, did not yield a clear picture and the exact metabolic consequences are not fully understood. Results In this work, deletion of the pyk gene, encoding pyruvate kinase, was carried out in the lysine-producing strain C. glutamicum lysCfbr, expressing a feedback resistant aspartokinase, to investigate the cellular response to deletion of this central glycolytic enzyme. Pyk deletion was achieved by allelic replacement, verified by PCR analysis and the lack of in vitro enzyme activity. The deletion mutant showed an overall growth behavior (specific growth rate, glucose uptake rate, biomass yield which was very similar to that of the parent strain, but differed in slightly reduced lysine formation, increased formation of the overflow metabolites dihydroxyacetone and glycerol and in metabolic fluxes around the pyruvate node. The latter involved a flux shift from pyruvate carboxylase (PC to PEPC, by which the cell maintained anaplerotic supply of the TCA cycle. This created a metabolic by-pass from PEP to pyruvate via malic enzyme demonstrating its contribution to metabolic flexibility of C. glutamicum on glucose. Conclusion The metabolic

  14. Prediction of DtxR regulon: Identification of binding sites and operons controlled by Diphtheria toxin repressor in Corynebacterium diphtheriae

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    Hasnain Seyed

    2004-09-01

    Full Text Available Abstract Background The diphtheria toxin repressor, DtxR, of Corynebacterium diphtheriae has been shown to be an iron-activated transcription regulator that controls not only the expression of diphtheria toxin but also of iron uptake genes. This study aims to identify putative binding sites and operons controlled by DtxR to understand the role of DtxR in patho-physiology of Corynebacterium diphtheriae. Result Positional Shannon relative entropy method was used to build the DtxR-binding site recognition profile and the later was used to identify putative regulatory sites of DtxR within C. diphtheriae genome. In addition, DtxR-regulated operons were also identified taking into account the predicted DtxR regulatory sites and genome annotation. Few of the predicted motifs were experimentally validated by electrophoretic mobility shift assay. The analysis identifies motifs upstream to the novel iron-regulated genes that code for Formamidopyrimidine-DNA glycosylase (FpG, an enzyme involved in DNA-repair and starvation inducible DNA-binding protein (Dps which is involved in iron storage and oxidative stress defense. In addition, we have found the DtxR motifs upstream to the genes that code for sortase which catalyzes anchoring of host-interacting proteins to the cell wall of pathogenic bacteria and the proteins of secretory system which could be involved in translocation of various iron-regulated virulence factors including diphtheria toxin. Conclusions We have used an in silico approach to identify the putative binding sites and genes controlled by DtxR in Corynebacterium diphtheriae. Our analysis shows that DtxR could provide a molecular link between Fe+2-induced Fenton's reaction and protection of DNA from oxidative damage. DtxR-regulated Dps prevents lethal combination of Fe+2 and H2O2 and also protects DNA by nonspecific DNA-binding. In addition DtxR could play an important role in host interaction and virulence by regulating the levels of sortase

  15. A novel bioremediation strategy for petroleum hydrocarbon pollutants using salt tolerant Corynebacterium variabile HRJ4 and biochar.

    Science.gov (United States)

    Zhang, Hairong; Tang, Jingchun; Wang, Lin; Liu, Juncheng; Gurav, Ranjit Gajanan; Sun, Kejing

    2016-09-01

    The present work aimed to develop a novel strategy to bioremediate the petroleum hydrocarbon contaminants in the environment. Salt tolerant bacterium was isolated from Dagang oilfield, China and identified as Corynebacterium variabile HRJ4 based on 16S rRNA gene sequence analysis. The bacterium had a high salt tolerant capability and biochar was developed as carrier for the bacterium. The bacteria with biochar were most effective in degradation of n-alkanes (C16, C18, C19, C26, C28) and polycyclic aromatic hydrocarbons (NAP, PYR) mixture. The result demonstrated that immobilization of C. variabile HRJ4 with biochar showed higher degradation of total petroleum hydrocarbons (THPs) up to 78.9% after 7-day of incubation as compared to the free leaving bacteria. The approach of this study will be helpful in clean-up of petroleum-contamination in the environments through bioremediation process using eco-friendly and cost effective materials like biochar. PMID:27593267

  16. Conjugative plasmid in Corynebacterium flaccumfaciens subsp. oortii that confers resistance to arsenite, arsenate, and antimony(III)

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    Hendrick, C.A.; Haskins, W.P.; Vidaver, A.K.

    1984-07-01

    Gene transfer systems for phytopathogenic corynebacteria have not been reported previously. In this paper a conjugative 46-megadalton plasmid (pDG101) found in Corynebacterium flaccumfaciens subsp. oorii CO101 is described that mediates resistance to arsenite, arsenate, and antimony(III). Transfer of the plasmid from CO101 to four other strains from the C. flaccumfaciens group occurred between cells immobilized on nitrocellulose filters or on agar surfaces. Transconjugant strains expressed the same levels of metal resistance as the donor strain and were able to act as donor strains in subsequent matings. The physical presence of the plasmid was detected by agarose gel electrophoresis. Arsenite-sensitive derivatives of the donor and transconjugant strains were obtained after heat treatment; these were cured of pDG101.

  17. Growth response of Avena sativa in amino-acids-rich soils converted from phenol-contaminated soils by Corynebacterium glutamicum.

    Science.gov (United States)

    Lee, Soo Youn; Kim, Bit-Na; Choi, Yong Woo; Yoo, Kye Sang; Kim, Yang-Hoon; Min, Jiho

    2012-04-01

    The biodegradation of phenol in laboratory-contaminated soil was investigated using the Gram-positive soil bacterium Corynebacterium glutamicum. This study showed that the phenol degradation caused by C. glutamicum was greatly enhanced by the addition of 1% yeast extract. From the toxicity test using Daphnia magna, the soil did not exhibit any hazardous effects after the phenol was removed using C. glutamicum. Additionally, the treatment of the phenolcontaminated soils with C. glutamicum increased various soil amino acid compositions, such as glycine, threonine, isoleucine, alanine, valine, leucine, tyrosine, and phenylalanine. This phenomenon induced an increase in the seed germination rate and the root elongation of Avena sativa (oat). This probably reflects that increased soil amino acid composition due to C. glutamicum treatment strengthens the plant roots. Therefore, the phenol-contaminated soil was effectively converted through increased soil amino acid composition, and additionally, the phenol in the soil environment was biodegraded by C. glutamicum.

  18. Draft Genome Sequence for the Type Strain of Corynebacterium afermentans LCDC 88-0199T, Isolated from a Human Blood Culture.

    Science.gov (United States)

    Bernier, Anne-Marie; Bernard, Kathryn

    2016-01-01

    A draft genome for Corynebacterium afermentans LCDC 88-0199(T) was investigated. The size of the genome was 2,345,615 bp with an observed G+C content of 64.85%. Annotation revealed 2 rRNA sequences, 54 tRNA genes, and 2,164 coding sequences. Genome coverage was 85× and consisted of 24 contigs with an N50 of 187,988 bp.

  19. Whole-Genome Sequence of Corynebacterium auriscanis Strain CIP 106629 Isolated from a Dog with Bilateral Otitis from the United Kingdom

    Science.gov (United States)

    Tiwari, Sandeep; Jamal, Syed Babar; Oliveira, Leticia Castro; Clermont, Dominique; Bizet, Chantal; Mariano, Diego; de Carvalho, Paulo Vinicius Sanches Daltro; Souza, Flavia; Pereira, Felipe Luiz; de Castro Soares, Siomar; Dorella, Fernanda; Carvalho, Alex; Leal, Carlos; Barh, Debmalya; Figueiredo, Henrique; Hassan, Syed Shah; Azevedo, Vasco

    2016-01-01

    In this work, we describe a set of features of Corynebacterium auriscanis CIP 106629 and details of the draft genome sequence and annotation. The genome comprises a 2.5-Mbp-long single circular genome with 1,797 protein-coding genes, 5 rRNA, 50 tRNA, and 403 pseudogenes, with a G+C content of 58.50%. PMID:27516502

  20. In Vitro Activities of the New Semisynthetic Glycopeptide Telavancin (TD-6424), Vancomycin, Daptomycin, Linezolid, and Four Comparator Agents against Anaerobic Gram-Positive Species and Corynebacterium spp.

    OpenAIRE

    Goldstein, Ellie J. C.; Citron, Diane M.; Merriam, C. Vreni; Warren, Yumi A.; Tyrrell, Kerin L.; Fernandez, Helen T.

    2004-01-01

    Telavancin is a new semisynthetic glycopeptide anti-infective with multiple mechanisms of action, including inhibition of bacterial membrane phospholipid synthesis and inhibition of bacterial cell wall synthesis. We determined the in vitro activities of telavancin, vancomycin, daptomycin, linezolid, quinupristin-dalfopristin, imipenem, piperacillin-tazobactam, and ampicillin against 268 clinical isolates of anaerobic gram-positive organisms and 31 Corynebacterium strains using agar dilution m...

  1. Draft Genome Sequence for the Type Strain of Corynebacterium afermentans LCDC 88-0199T, Isolated from a Human Blood Culture

    Science.gov (United States)

    Bernier, Anne-Marie

    2016-01-01

    A draft genome for Corynebacterium afermentans LCDC 88-0199T was investigated. The size of the genome was 2,345,615 bp with an observed G+C content of 64.85%. Annotation revealed 2 rRNA sequences, 54 tRNA genes, and 2,164 coding sequences. Genome coverage was 85× and consisted of 24 contigs with an N50 of 187,988 bp. PMID:27389276

  2. Molecular cloning and DNA sequence analysis of a diphtheria tox iron-dependent regulatory element (dtxR) from Corynebacterium diphtheriae.

    OpenAIRE

    Boyd, J.; Oza, M N; Murphy, J. R.

    1990-01-01

    Although the structural gene for diphtheria toxin, tox, is carried by a family of closely related corynebacteriophages, the regulation of tox expression is controlled, to a large extent, by its bacterial host Corynebacterium diphtheriae. Optimal yields of tox gene products are obtained only when iron becomes the growth-rate-limiting substrate. Previous studies suggest that regulation of tox expression is mediated through an iron-binding aporepressor. To facilitate molecular cloning of the tox...

  3. Effects of non-specific immunopotentiators in experimental Schistosoma mansoni infection: II. Corynebacterium parvum Efeitos de imunopotenciadores não específicos na infecção experimental pelo Schistosoma mansoni: II. Corynebacterium parvum

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    Kirte M Teixeira

    1996-10-01

    Full Text Available The effects of Corynebacterium parvum on host protection, tissue reaction and "in vivo" chemotaxis in Schistosoma mansoni infected mice were studied. The C. parvum was given intraperitoneally using a dose of 0.7 mg, twice a week (for 4 weeks, thirty days before (prophylactic treatment or after infection (curative treatment. The host protection was evaluated through the recovery of adult worms by liver perfusion and was lower in the prophylactic group as compared to the control group (p = 0.018, resulting in 44% protection. The "in vivo" leukocyte response in both prophylactic and curative groups was higher as compared to the infected/non treated group (p = 0.009 and p = 0.003, respectively. Tissue reactions were described in the experimental and control groups, but there were not remarkable differences among them. The possible biological implications and relevance of the findings for the defensive response of the host and control of schistosomiasis are discussed.Neste trabalho foram avaliados os efeitos do Corynebacterium parvum na proteção do hospedeiro, reação tecidual e quimiotaxia "in vivo" em camundongos infectados pelo S. mansoni. O C. parvum foi dado intraperitonealmente usando uma dose de 0,7 mg, duas vezes por semana (durante 04 semanas, 30 dias antes (tratamento profilático e 30 dias após a infecção (tratamento curativo. A proteção do hospedeiro foi avaliada através da contagem de vermes adultos obtidos através da perfusão hepática de camundongos infectados e esse número foi bem menor no grupo profilático comparado ao grupo controle (p = 0,018, obtendo-se 44% de proteção. A resposta quimiotática "in vivo", nos grupos curativo e profilático, foi maior do que no grupo infectado/não tratado (p = 0,009 e p = 0,003, respectivamente. As reações teciduais foram descritas em todos os grupos, embora não tenha ocorrido diferenças marcantes entre eles. As possíveis implicações biológicas e a relevância dos achados

  4. Characterization of OxyR as a negative transcriptional regulator that represses catalase production in Corynebacterium diphtheriae.

    Science.gov (United States)

    Kim, Ju-Sim; Holmes, Randall K

    2012-01-01

    Corynebacterium diphtheriae and Corynebacterium glutamicum each have one gene (cat) encoding catalase. In-frame Δcat mutants of C. diphtheriae and C. glutamicum were hyper-sensitive to growth inhibition and killing by H(2)O(2). In C. diphtheriae C7(β), both catalase activity and cat transcription decreased ~2-fold during transition from exponential growth to early stationary phase. Prototypic OxyR in Escherichia coli senses oxidative stress and it activates katG transcription and catalase production in response to H(2)O(2). In contrast, exposure of C. diphtheriae C7(β) to H(2)O(2) did not stimulate transcription of cat. OxyR from C. diphtheriae and C. glutamicum have 52% similarity with E. coli OxyR and contain homologs of the two cysteine residues involved in H(2)O(2) sensing by E. coli OxyR. In-frame ΔoxyR deletion mutants of C. diphtheriae C7(β), C. diphtheriae NCTC13129, and C. glutamicum were much more resistant than their parental wild type strains to growth inhibition by H(2)O(2). In the C. diphtheriae C7(β) ΔoxyR mutant, cat transcripts were about 8-fold more abundant and catalase activity was about 20-fold greater than in the C7(β) wild type strain. The oxyR gene from C. diphtheriae or C. glutamicum, but not from E. coli, complemented the defect in ΔoxyR mutants of C. diphtheriae and C. glutamicum and decreased their H(2)O(2) resistance to the level of their parental strains. Gel-mobility shift, DNaseI footprint, and primer extension assays showed that purified OxyR from C. diphtheriae C7(β) bound, in the presence or absence of DTT, to a sequence in the cat promoter region that extends from nucleotide position -55 to -10 with respect to the +1 nucleotide in the cat ORF. These results demonstrate that OxyR from C. diphtheriae or C. glutamicum functions as a transcriptional repressor of the cat gene by a mechanism that is independent of oxidative stress induced by H(2)O(2). PMID:22438866

  5. Characterization of OxyR as a negative transcriptional regulator that represses catalase production in Corynebacterium diphtheriae.

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    Ju-Sim Kim

    Full Text Available Corynebacterium diphtheriae and Corynebacterium glutamicum each have one gene (cat encoding catalase. In-frame Δcat mutants of C. diphtheriae and C. glutamicum were hyper-sensitive to growth inhibition and killing by H(2O(2. In C. diphtheriae C7(β, both catalase activity and cat transcription decreased ~2-fold during transition from exponential growth to early stationary phase. Prototypic OxyR in Escherichia coli senses oxidative stress and it activates katG transcription and catalase production in response to H(2O(2. In contrast, exposure of C. diphtheriae C7(β to H(2O(2 did not stimulate transcription of cat. OxyR from C. diphtheriae and C. glutamicum have 52% similarity with E. coli OxyR and contain homologs of the two cysteine residues involved in H(2O(2 sensing by E. coli OxyR. In-frame ΔoxyR deletion mutants of C. diphtheriae C7(β, C. diphtheriae NCTC13129, and C. glutamicum were much more resistant than their parental wild type strains to growth inhibition by H(2O(2. In the C. diphtheriae C7(β ΔoxyR mutant, cat transcripts were about 8-fold more abundant and catalase activity was about 20-fold greater than in the C7(β wild type strain. The oxyR gene from C. diphtheriae or C. glutamicum, but not from E. coli, complemented the defect in ΔoxyR mutants of C. diphtheriae and C. glutamicum and decreased their H(2O(2 resistance to the level of their parental strains. Gel-mobility shift, DNaseI footprint, and primer extension assays showed that purified OxyR from C. diphtheriae C7(β bound, in the presence or absence of DTT, to a sequence in the cat promoter region that extends from nucleotide position -55 to -10 with respect to the +1 nucleotide in the cat ORF. These results demonstrate that OxyR from C. diphtheriae or C. glutamicum functions as a transcriptional repressor of the cat gene by a mechanism that is independent of oxidative stress induced by H(2O(2.

  6. The identification of enzyme targets for the optimization of a valine producing Corynebacterium glutamicum strain using a kinetic model.

    Science.gov (United States)

    Magnus, Jørgen Barsett; Oldiges, Marco; Takors, Ralf

    2009-01-01

    The enzyme targets for the rational optimization of a Corynebacterium glutamicum strain constructed for valine production are identified by analyzing the control of flux in the valine/leucine pathway. The control analysis is based on measurements of the intracellular metabolite concentrations and on a kinetic model of the reactions in the investigated pathway. Data-driven and model-based methods are used and evaluated against each other. The approach taken gives a quantitative evaluation of the flux control and it is demonstrated how the understanding of flux control is used to reach specific recommendations for strain optimization. The flux control coefficients (FCCs) with respect to the valine excretion rate were calculated, and it was found that the control is distributed mainly between the acetohydroxyacid synthase enzyme (FCC = 0.32), the branched chain amino acid transaminase (FCC = 0.27), and the exporting translocase (FCC = 0.43). The availability of the precursor pyruvate has substantial influence on the valine flux, whereas the cometabolites are less important as demonstrated by the calculation of the respective response coefficients. The model is further used to make in-silico predictions of the change in valine flux following a change in enzyme level. A doubling of the enzyme level of valine translocase will result in an increase in valine flux of 31%. By optimizing the enzyme levels with respect to valine flux it was found that the valine flux can be increased by a factor 2.5 when the optimal enzyme levels are implemented.

  7. Enhanced valine production in Corynebacterium glutamicum with defective H+-ATPase and C-terminal truncated acetohydroxyacid synthase.

    Science.gov (United States)

    Wada, Masaru; Hijikata, Nowaki; Aoki, Ryo; Takesue, Nobuchika; Yokota, Atsushi

    2008-11-01

    We have reported increased glutamate production by a mutant of Corynebacterium glutamicum ATCC14067 (strain F172-8) with reduced H(+)-ATPase activity under biotin-limiting culture conditions (Aoki et al. Biosci. Biotechnol. Biochem., 69, 1466-1472 (2005)). In the present study, we examined valine production by an H(+)-ATPase-defective mutant of C. glutamicum. Using the double-crossover chromosome replacement technique, we constructed a newly defined H(+)-ATPase-defective mutant from ATCC13032. After transforming the new strain (A-1) with a C-terminal truncation of acetohydroxyacid synthase gene (ilvBN), valine production increased from 21.7 mM for the wild-type strain to 46.7 mM for the A-1 in shaking flask cultures with 555 mM glucose. Increased production of the valine intermediate acetoin was also observed in A-1, and was reduced by inserting acetohydroxyacid isomeroreductase gene (ilvC) into the ilvBN plasmid. After transformation with this new construct, valine production increased from 38.3 mM for the wild-type strain to 95.7 mM for A-1 strain. To the best of our knowledge, this is the first report indicating that an H(+)-ATPase-defective mutant of C. glutamicum is capable of valine production. Our combined results with glutamate and valine suggest that the H(+)-ATPase defect is also effective in the fermentative production of other practical compounds.

  8. The physiological role of riboflavin transporter and involvement of FMN-riboswitch in its gene expression in Corynebacterium glutamicum.

    Science.gov (United States)

    Takemoto, Norihiko; Tanaka, Yuya; Inui, Masayuki; Yukawa, Hideaki

    2014-05-01

    Riboflavin is a precursor of flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), which work as cofactors of numerous enzymes. Understanding the supply system of these cofactors in bacteria, particularly those used for industrial production of value added chemicals, is important given the pivotal role the cofactors play in substrate metabolism. In this work, we examined the effect of disruption of riboflavin utilization genes on cell growth, cytoplasmic flavin levels, and expression of riboflavin transporter in Corynebacterium glutamicum. Disruption of the ribA gene that encodes bifunctional GTP cyclohydrolase II/3,4-dihydroxy-2-butanone 4-phosphate synthase in C. glutamicum suppressed growth in the absence of supplemental riboflavin. The growth was fully recovered upon supplementation with 1 μM riboflavin, albeit at reduced intracellular concentrations of FMN and FAD during the log phase. Concomitant disruption of the ribA and ribM gene that encodes a riboflavin transporter exacerbated supplemental riboflavin requirement from 1 μM to 50 μM. RibM expression in FMN-rich cells was about 100-fold lower than that in FMN-limited cells. Mutations in putative FMN-riboswitch present immediately upstream of the ribM gene abolished the FMN response. This 5'UTR sequence of ribM constitutes a functional FMN-riboswitch in C. glutamicum. PMID:24531272

  9. Effects of the removal of the primary tumor and immunotherapy with Corynebacterium parvum on metastatic tumor proliferation.

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    Hashimoto,Osamu

    1983-08-01

    Full Text Available The effects of surgical intervention by removal of the primary focus, and the effectiveness of an immunomodulator, Corynebacterium parvum (Cp, on the proliferation of metastatic tumor tissue were investigated by following the postoperative changes in the 3H-thymidine labelling rate of metastatic tissue in an experimental model of metastasis in mice. In addition, the delayed type hypersensitivity reaction (DTH was studied to investigate the immune capacity of the host. The labelling rate of mice that had the primary focus removed remained high with little variation, while that of the mice not operated on decreased gradually. On the other hand, in mice undergoing a sham operation, the rate was the same as that of the mice with the primary focus removed for a short while, but then gradually decreased. When Cp was administered, especially before removal of the primary focus, the rate was lower than that of the tumor bearing control group and decreased steadily. The number of pulmonary metastatic nodules was increased by removal of the primary focus, but this increase was inhibited by the administration of Cp which prolonged life. The depression in the DTH was less in the group given Cp preoperativeLy than in either the group of mice having the primary focus removed or those not having it removed.

  10. Combined use of local irradiation and corynebacterium parvum in the treatment of the murine line 1 lung carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Ullrich, R.L.; Adams, L.M.

    1978-02-01

    The effectiveness of Corynebacterium parvum in combination with local irradiation has been examined in the treatment of the murine line 1 lung carcinoma, a highly radioresistant, weakly immunogenic tumor that kills the host by means of metastatic spread. Sixteen-week-old, specific-pathogen-free female BABL/c mice were given 10/sup 6/ tumor cells im into the right thigh. Tumors were irradiated on Day 7 after transplant. Those receiving C. parvum treatment were given 0.1 mg either by the intralesional (il), ip, or iv route on Day 4 after transplant or by the il or ip route on Day 8. An additional group received C. parvum ip once a week for 4 weeks beginning on Day 8. The influence of the various treatments on local control and metastasis was assessed. To evaluate further the time course and incidence of metastases, cleared lungs were examined at 21, 28, and 35 days in groups given irradiation combined with C. parvum on day 8. C. parvum was more effective in facilitating local control and inhibiting metastatic spread when given after radiation exposure rather than before.

  11. Improvement of enantioselectivity of the B-type halohydrin hydrogen-halide-lyase from Corynebacterium sp. N-1074.

    Science.gov (United States)

    Watanabe, Fumiaki; Yu, Fujio; Ohtaki, Akashi; Yamanaka, Yasuaki; Noguchi, Keiichi; Odaka, Masafumi; Yohda, Masafumi

    2016-09-01

    Halohydrin hydrogen-halide-lyase (H-Lyase) is a bacterial enzyme involved in the degradation of halohydrins. This enzyme catalyzes the intramolecular nucleophilic displacement of a halogen by a vicinal hydroxyl group in halohydrins, producing the corresponding epoxides. The H-Lyases have been classified into A, B and C subtypes based on amino acid sequence similarities. These enzymes have attracted much attention as industrial catalysts in the synthesis of chiral chemicals from prochiral halohydrins. In the present study, we constructed mutants of B-type H-Lyase from Corynebacterium sp. N-1074 (HheB) displaying higher enantioselectivity by structure-based site-directed mutagenesis and random mutagenesis. A triple mutant of HheB exhibited 98.5% enantioselectivity, the highest ever reported, toward (R)-4-chloro-3-hydroxy-butyronitrile production, with the yield reaching approximately two-fold that of the wild-type enzyme. We discuss the structural basis of the high enantioselectivity and productivity of the mutant by comparing the crystal structures of the mutant HheB and the wild-type enzyme in complex with or without the substrate analogue. PMID:27215832

  12. Corynebacterium glutamicum harbours a molybdenum cofactor-dependent formate dehydrogenase which alleviates growth inhibition in the presence of formate.

    Science.gov (United States)

    Witthoff, Sabrina; Eggeling, Lothar; Bott, Michael; Polen, Tino

    2012-09-01

    Here, we show that Corynebacterium glutamicum ATCC 13032 co-metabolizes formate when it is grown with glucose as the carbon and energy source. CO(2) measurements during bioreactor cultivation and use of (13)C-labelled formate demonstrated that formate is almost completely oxidized to CO(2). The deletion of fdhF (cg0618), annotated as formate dehydrogenase (FDH) and located in a cluster of genes conserved in the family Corynebacteriaceae, prevented formate utilization. Similarly, deletion of fdhD (cg0616) resulted in the inability to metabolize formate and deletion of cg0617 markedly reduced formate utilization. These results illustrated that all three gene products are required for FDH activity. Growth studies with molybdate and tungstate indicated that the FDH from C. glutamicum ATCC 13032 is a molybdenum-dependent enzyme. The presence of 100 mM formate caused a 25 % lowered growth rate during cultivation of C. glutamicum ATCC 13032 wild-type in glucose minimal medium. This inhibitory effect was increased in the strains lacking FDH activity. Our data demonstrate that C. glutamicum ATCC 13032 possesses an FDH with a currently unknown electron acceptor. The presence of the FDH might help the soil bacterium C. glutamicum ATCC 13032 to alleviate growth retardation caused by formate, which is ubiquitously present in the environment. PMID:22767548

  13. Improvement of enantioselectivity of the B-type halohydrin hydrogen-halide-lyase from Corynebacterium sp. N-1074.

    Science.gov (United States)

    Watanabe, Fumiaki; Yu, Fujio; Ohtaki, Akashi; Yamanaka, Yasuaki; Noguchi, Keiichi; Odaka, Masafumi; Yohda, Masafumi

    2016-09-01

    Halohydrin hydrogen-halide-lyase (H-Lyase) is a bacterial enzyme involved in the degradation of halohydrins. This enzyme catalyzes the intramolecular nucleophilic displacement of a halogen by a vicinal hydroxyl group in halohydrins, producing the corresponding epoxides. The H-Lyases have been classified into A, B and C subtypes based on amino acid sequence similarities. These enzymes have attracted much attention as industrial catalysts in the synthesis of chiral chemicals from prochiral halohydrins. In the present study, we constructed mutants of B-type H-Lyase from Corynebacterium sp. N-1074 (HheB) displaying higher enantioselectivity by structure-based site-directed mutagenesis and random mutagenesis. A triple mutant of HheB exhibited 98.5% enantioselectivity, the highest ever reported, toward (R)-4-chloro-3-hydroxy-butyronitrile production, with the yield reaching approximately two-fold that of the wild-type enzyme. We discuss the structural basis of the high enantioselectivity and productivity of the mutant by comparing the crystal structures of the mutant HheB and the wild-type enzyme in complex with or without the substrate analogue.

  14. Two-stage pH Control Mode in Batch Fermentation of a Novel Bioflocculant from Corynebacterium Glutamicum

    Institute of Scientific and Technical Information of China (English)

    HE Ning; WU Xiao-jie; DENG Xu; LU Ying-hua; LI Qing-biao

    2004-01-01

    The effect of pH of the fermentation medium on cell growth and the production of a novel bioflocculant (named REA-11 ) by Corynebacterium glutamicum CCTCC M201005 were investigated. The maximum biomass (2.23 g/L) and flocculating activity (142.2 U/mL) were simultaneously obtained at the 14th hour when the pH value of the culture medium was maintained at 7.0 during the whole fermentation process. The production of REA-11 kept on a trend of increase till the later phase of fermentation process, which resulted in the ultimate flocculating activity of the culture broth to enhance to nearly 100 U/mL at pH 6.0. A twostage pH control mode was adopted in REA-11 production in which the pH value of the culture medium was controlled at 7.0 during the first 14 h, then decreased to 6.0 that was maintained until the end of the fermentation process. With the two-stage pH control mode, the maximum flocculating activity reached 178.8 U/mL which was 30% higher than that obtained under the condition of pH 7.0 and the biomass enhanced about 15%. Compared with the fermentation process without pH control, REA-11 production and cell growth via the two-stage pH control mode increased 80% and 25%, respectively.

  15. Formation of xylitol and xylitol-5-phosphate and its impact on growth of d-xylose-utilizing Corynebacterium glutamicum strains.

    Science.gov (United States)

    Radek, Andreas; Müller, Moritz-Fabian; Gätgens, Jochem; Eggeling, Lothar; Krumbach, Karin; Marienhagen, Jan; Noack, Stephan

    2016-08-10

    Wild-type Corynebacterium glutamicum has no endogenous metabolic activity for utilizing the lignocellulosic pentose d-xylose for cell growth. Therefore, two different engineering approaches have been pursued resulting in platform strains harbouring a functional version of either the Isomerase (ISO) or the Weimberg (WMB) pathway for d-xylose assimilation. In a previous study we found for C. glutamicum WMB by-product formation of xylitol during growth on d-xylose and speculated that the observed lower growth rates are due to the growth inhibiting effect of this compound. Based on a detailed phenotyping of the ISO, WMB and the wild-type strain of C. glutamicum, we here show that this organism has a natural capability to synthesize xylitol from d-xylose under aerobic cultivation conditions. We furthermore observed the intracellular accumulation of xylitol-5-phosphate as a result of the intracellular phosphorylation of xylitol, which was particularly pronounced in the C. glutamicum ISO strain. Interestingly, low amounts of supplemented xylitol strongly inhibit growth of this strain on d-xylose, d-glucose and d-arabitol. These findings demonstrate that xylitol is a suitable substrate of the endogenous xylulokinase (XK, encoded by xylB) and its overexpression in the ISO strain leads to a significant phosphorylation of xylitol in C. glutamicum. Therefore, in order to circumvent cytotoxicity by xylitol-5-phosphate, the WMB pathway represents an interesting alternative route for engineering C. glutamicum towards efficient d-xylose utilization. PMID:27297548

  16. A role for sigma factor SigE in Corynebacterium pseudotuberculosis resistance to nitric oxide/ peroxide stress

    Directory of Open Access Journals (Sweden)

    Luis G. C. Pacheco

    2012-04-01

    Full Text Available Pathogenic intracellular bacteria can respond to antimicrobial mechanisms of the host cell through transient activation of stress-responsive genes by alternative sigma (σ factors of the RNA polymerase. We evaluated the contribution of the extracytoplasmic function sigma factor σE for Corynebacterium pseudotuberculosis resistance to stress conditions resembling those found intracellularly during infection. A sigE null mutant strain (delta-sigE of this bacterium was more susceptible in vitro to acidic pH, cell surface stressors, and biologically relevant concentrations of nitric oxide (NO. The same mutant strain was unable to persist in C57BL/6 mice but remained infective in mice lacking inducible nitric oxide synthase (iNOS, confirming the significance of σE for resistance to nitric oxide/peroxide stress in vivo. High-throughput proteomic analysis identified NO-responsive extracellular proteins of C. pseudotuberculosis and demonstrated the participation of σE in composition of this bacterium´s exoproteome.

  17. Novel Polyoxyethylene-Containing Glycolipids Are Synthesized in Corynebacterium matruchotii and Mycobacterium smegmatis Cultured in the Presence of Tween 80

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    Cindy Wang

    2011-01-01

    Full Text Available The addition of polyoxyethylene sorbitan monooleate (Tween 80 to a culture of mycobacteria greatly influences cell permeability and sensitivity to antibiotics but very little is known regarding the underlying mechanism. Here we show that Corynebacterium matruchotii (surrogate of mycobacteria converts Tween 80 to a structural series of polyoxyethylenic acids which are then used to form novel series-2A and series-2B glycolipids. Minor series-3 glycolipids were also synthesized. The polyoxyethylenic acids replaced corynomycolic acids in the cell wall. Correspondingly the trehalose dicorynomycolate content was reduced. MALDI mass spectrometry, MS-MS, 1H-NMR, and 13C-NMR were used to characterize the series-2 glycolipids. Series-2A glycolipid is trehalose 6-C36:2-corynomycolate-6′-polyoxyethylenate and series-2B glycolipid is trehalose 6-C36:2-corynomycolate-6′-furan ring-containing polyoxyethylenate. Mycobacterium smegmatis grown in the presence of Tween 80 also synthesizes series-2 type glycolipids. The synthesis of these novel glycolipids in corynebacteria and mycobacteria should result in gross changes in the cell wall permeability and drug sensitivity.

  18. Interaction between DAHP synthase and chorismate mutase endows new regulation on DAHP synthase activity in Corynebacterium glutamicum.

    Science.gov (United States)

    Li, Pan-Pan; Li, De-Feng; Liu, Di; Liu, Yi-Ming; Liu, Chang; Liu, Shuang-Jiang

    2013-12-01

    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 CMCg, prephenate markedly inhibited DSCg activity. Prephenate competed with the substrate phosphoenolpyruvate, and the inhibition constant (K i) was determined to be 0.945 mM. Modeling based on the structure of the complex formed between DAHP synthase and chorismate mutase of Mycobacterium tuberculosis predicted the interaction surfaces of the putative DSCg-CMCg complex. The amino acid residues and structural domains that contributed to the interaction surfaces were experimentally identified to be the (212)SPAGARYE(219) sequence of DSCg and the (60)SGGTR(64) loop and C-terminus ((97)RGKLG(101)) of CMCg. PMID:23467831

  19. Ethambutol-mediated cell wall modification in recombinant Corynebacterium glutamicum increases the biotransformation rates of cyclohexanone derivatives.

    Science.gov (United States)

    Yun, Ji-Yeong; Lee, Jung-Eun; Yang, Kyung-Mi; Cho, Suekyung; Kim, Arim; Kwon, Yong-Uk; Kwon, Yong-Euk; Park, Jin-Byung

    2012-01-01

    The effects of structural modification of cell wall on the biotransformation capability by recombinant Corynebacterium glutamicum cells, expressing the chnB gene encoding cyclohexanone monooxygenase of Acinetobacter calcoaceticus NCIMB 9871, were investigated. Baeyer-Villiger oxygenation of 2-(2'-acetoxyethyl) cyclohexanone (MW 170 Da) into R-7-(2'-acetoxyethyl)-2-oxepanone was used as a model reaction. The whole-cell biotransformation followed Michaelis-Menten kinetics. The V (max) and K (S) values were estimated as 96.8 U g(-1) of dry cells and 0.98 mM, respectively. The V (max) was comparable with that of cyclohexanone oxygenation, whereas the K (S) was almost eightfold higher. The K (S) value of 2-(2'-acetoxyethyl) cyclohexanone oxygenation was reduced by ca. 30% via altering the cell envelop structure of C. glutamicum with ethambutol, which inhibits arabinosyl transferases involved in the biosynthesis of cell wall arabinogalactan and mycolate layers. The higher whole-cell biotransformation rate was also observed in the oxygenation of ethyl 2-cyclohexanone acetate upon ethambutol treatment of the recombinant C. glutamicum. Therefore, it was assumed that the biotransformation efficiency of C. glutamicum-based biocatalysts, with respect to medium- to large-sized lipophilic organic substrates (MW > ca. 170), can be enhanced by engineering their cell wall outer layers, which are known to function as a formidable barrier to lipophilic molecules.

  20. Phenotypic, molecular characterization, antimicrobial susceptibility and draft genome sequence of Corynebacterium argentoratense strains isolated from clinical samples

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    I. Fernández-Natal

    2016-03-01

    Full Text Available During a 12-year period we isolated five Corynebacterium argentoratense strains identified by phenotypic methods, including the use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF and 16S rRNA gene sequencing. In addition, antimicrobial susceptibility was determined, and genome sequencing for the detection of antibiotic resistance genes was performed. The organisms were isolated from blood and throat cultures and could be identified by all methods used. All strains were resistant to cotrimoxazole, and resistance to β-lactams was partly present. Two strains were resistant to erythromycin and clindamycin. The draft genome sequences of theses isolates revealed the presence of the erm(X resistance gene that is embedded in the genetic structure of the transposable element Tn5423. Although rarely reported as a human pathogen, C. argentoratense can be involved in bacteraemia and probably in other infections. Our results also show that horizontal transfer of genes responsible for antibiotic resistance is occurring in this species.

  1. PARÁMETROS FISICOQUÍMICOS PARA LA SÍNTESIS DE ÁCIDO LÁCTICO O ETANOL DE LA BACTERIA (Corynebacterium glutamicum Physico-Chemical Parameter for Production of Lactic Acid or Ethanol of (Corynebacterium glutamicum Bacteria

    Directory of Open Access Journals (Sweden)

    ANGÉLICA CASTELLANOS

    2011-08-01

    Full Text Available El interés por obtener productos para la industria de biocombustibles a partir de desechos agrícolas, conduce a la búsqueda de nuevos sistemas biotecnológicos resistentes y costo-efectivos. Corynebacterium glutamicum, es un microorganismo usado para producir amino-ácidos que crece en gran variedad de sustratos y es resistente durante la fermentación, a variaciones de pH, temperatura, presión osmótica y acumulación de alcohol, características que lo hacen candidato a ser mejorado para la síntesis de ácido láctico y etanol. Aún se desconocen aspectos de su fisiología que aumenten su eficiencia en convertir azúcares (C5 y C6 en estos dos metabolitos. Por tanto, este trabajo buscó identificar los parámetros fisicoquímicos que tuvieron un mayor efecto sobre crecimiento bacteriano y síntesis de ácido láctico o etanol en un sistema por lotes. Para lograr este objetivo, ocho variables fueron evaluadas en un modelo estadístico producido en erlenmeyer; con los resultados obtenidos, se hallaron las mejores condiciones que fueron puestas a prueba en un cultivo en biorreactor. La temperatura, concentración de biotina y azúcar fueron las variables de mayor impacto (pThe interest to obtain products for the bio-fuel industry from renewable resources has directed research to find resistant and costs-effective biotechnological systems. Corynebacterium glutamicum, is a microorganism used to produce amino acids, that grows in wide variety of substrates and its resistance during fermentation to pH, temperature, osmotic pressure variations and alcohol aggregate, renders this organism a suitable candidate to improve by genetic modifications lactic acid and ethanol synthesis. However, some aspects of its physiology remain unknown, such us increase lactic acid and ethanol production from C5 and C6 sugars. For this reason, the main aim in our work was to identify the most important variables with impact on culture and the best culture conditions

  2. Technetium-99m labeling and fibronectin binding ability of Corynebacterium diphtheriae; Marcacao de Corynebacterium diphtheriae com Tecnecio-99m e avaliacao da capacidade de ligacao a fibronectina de plasma humano

    Energy Technology Data Exchange (ETDEWEB)

    Souza, S.M.S.; Nagao, P.E.; Bernardo-Filho, M. [Universidade do Estado do Rio de Janeiro, RJ (Brazil). Inst. de Biologia Roberto Alcantara Gomes; Pereira, G.A.; Napoleao, F.; Andrade, A.F.B.; Hirata Junior, R.; Mattos-Guaraldi, A.L. [Universidade do Estado do Rio de Janeiro, RJ (Brazil). Faculdade de Ciencias Medicas

    2004-04-15

    The use of radionuclides has permitted advances in areas of clinical and scientific knowledge. Several molecules and cells have been labelled with Technetium-99m ({sup 99m}Tc). The stannous chloride (SnCl{sub 2}) has a significant influence on the labeling and stability of {sup 99m}Tc radiotracers. The frequent risk of diphtheria epidemics has intensified interest in the virulence factors of Corynebacterium diphtheriae. Although studies have looked at potential adhesins including haemagglutinins and exposed sugar residues, the molecular basis of mechanisms of adherence remains unclear. Adherence of pathogens to mammalian tissues may be mediated by fibronectin (FN) found in body fluids, matrix of connective tissues, and cell surfaces. In the present study we evaluated the binding ability to human plasma FN by {sup 99m}Tc labeled-C.diphtheriae. Due to adverse effects of stannous ions, microorganisms were submitted to survival and filamentation induction assays. Data showed a dose dependent susceptibility to SnCl{sub 2} bactericidal effects. Cell filamentation was observed for concentrations of SnCl{sub 2} > 110 {mu}g/ml. Adherence levels of {sup 99m}Tc labelled 241strain to coverslips coated with 20 {mu}g/ml FN were higher (P = 0.0037) than coated with bovine serum albumin. FN binding by the sucrose fermenting 241 C. diphtheriae strain (8.9% + 2.6) was significantly lower (P=0.0139) than Staphylococcus aureus Cowan I strain (34.1% {+-} 1.2). Therefore, bacterial {sup 99m}Tc labeling represents an additional tool that may contribute to the comprehension of C. diphtheriae interactions with host receptors such as FN that act as biological organizers by holding bacterial cells in position and guiding their migration. (author)

  3. Transcriptional Regulation of the Vanillate Utilization Genes (vanABK Operon) of Corynebacterium glutamicum by VanR, a PadR-Like Repressor

    OpenAIRE

    Morabbi Heravi, Kambiz; Lange, Julian; Watzlawick, Hildegard; Kalinowski, Jörn; Altenbuchner, Josef

    2014-01-01

    Corynebacterium glutamicum is able to utilize vanillate, the product of lignin degradation, as the sole carbon source. The vanillate utilization components are encoded by the vanABK operon. The vanA and vanB genes encode the subunits of vanillate O-demethylase, converting vanillate to protocatechuate, while VanK is the specific vanillate transporter. The vanABK operon is regulated by a PadR-type repressor, VanR. Heterologous gene expression and variations of the vanR open reading frame reveal...

  4. Carbon Flux Analysis by 13C Nuclear Magnetic Resonance To Determine the Effect of CO2 on Anaerobic Succinate Production by Corynebacterium glutamicum

    OpenAIRE

    Radoš, Dušica; David L Turner; Fonseca, Luís L.; Carvalho, Ana Lúcia; Blombach, Bastian; Eikmanns, Bernhard J.; Neves, Ana Rute; Santos, Helena

    2014-01-01

    Wild-type Corynebacterium glutamicum produces a mixture of lactic, succinic, and acetic acids from glucose under oxygen deprivation. We investigated the effect of CO2 on the production of organic acids in a two-stage process: cells were grown aerobically in glucose, and subsequently, organic acid production by nongrowing cells was studied under anaerobic conditions. The presence of CO2 caused up to a 3-fold increase in the succinate yield (1 mol per mol of glucose) and about 2-fold increase i...

  5. 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

    Science.gov (United States)

    Scheele, Sandra; Oertel, Dan; Bongaerts, Johannes; Evers, Stefan; Hellmuth, Hendrik; Maurer, Karl-Heinz; Bott, Michael; Freudl, Roland

    2013-01-01

    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 demonstrate for the first time that, also for cofactor-containing proteins, a secretory production strategy is a feasible and promising alternative to conventional intracellular expression strategies. PMID:23163932

  6. 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.

    Science.gov (United States)

    Scheele, Sandra; Oertel, Dan; Bongaerts, Johannes; Evers, Stefan; Hellmuth, Hendrik; Maurer, Karl-Heinz; Bott, Michael; Freudl, Roland

    2013-03-01

    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 demonstrate for the first time that, also for cofactor-containing proteins, a secretory production strategy is a feasible and promising alternative to conventional intracellular expression strategies.

  7. Toward homosuccinate fermentation: metabolic engineering of Corynebacterium glutamicum for anaerobic production of succinate from glucose and formate.

    Science.gov (United States)

    Litsanov, Boris; Brocker, Melanie; Bott, Michael

    2012-05-01

    Previous studies have demonstrated the capability of Corynebacterium glutamicum for anaerobic succinate production from glucose under nongrowing conditions. In this work, we have addressed two shortfalls of this process, the formation of significant amounts of by-products and the limitation of the yield by the redox balance. To eliminate acetate formation, a derivative of the type strain ATCC 13032 (strain BOL-1), which lacked all known pathways for acetate and lactate synthesis (Δcat Δpqo Δpta-ackA ΔldhA), was constructed. Chromosomal integration of the pyruvate carboxylase gene pyc(P458S) into BOL-1 resulted in strain BOL-2, which catalyzed fast succinate production from glucose with a yield of 1 mol/mol and showed only little acetate formation. In order to provide additional reducing equivalents derived from the cosubstrate formate, the fdh gene from Mycobacterium vaccae, coding for an NAD(+)-coupled formate dehydrogenase (FDH), was chromosomally integrated into BOL-2, leading to strain BOL-3. In an anaerobic batch process with strain BOL-3, a 20% higher succinate yield from glucose was obtained in the presence of formate. A temporary metabolic blockage of strain BOL-3 was prevented by plasmid-borne overexpression of the glyceraldehyde 3-phosphate dehydrogenase gene gapA. In an anaerobic fed-batch process with glucose and formate, strain BOL-3/pAN6-gap accumulated 1,134 mM succinate in 53 h with an average succinate production rate of 1.59 mmol per g cells (dry weight) (cdw) per h. The succinate yield of 1.67 mol/mol glucose is one of the highest currently described for anaerobic succinate producers and was accompanied by a very low level of by-products (0.10 mol/mol glucose).

  8. Corynebacterium glutamicum ATP-phosphoribosyl transferases suitable for L-histidine production--Strategies for the elimination of feedback inhibition.

    Science.gov (United States)

    Kulis-Horn, Robert K; Persicke, Marcus; Kalinowski, Jörn

    2015-07-20

    L-Histidine biosynthesis in Corynebacterium glutamicum is mainly regulated by L-histidine feedback inhibition of the ATP-phosphoribosyltransferase HisG that catalyzes the first step of the pathway. The elimination of this feedback inhibition is the first and most important step in the development of an L-histidine production strain. For this purpose, a combined approach of random mutagenesis and rational enzyme redesign was performed. Mutants spontaneously resistant to the toxic L-histidine analog β-(2-thiazolyl)-DL-alanine (2-TA) revealed novel and unpredicted mutations in the C-terminal regulatory domain of HisG resulting in increased feedback resistance. Moreover, deletion of the entire C-terminal regulatory domain in combination with the gain of function mutation S143F in the catalytic domain resulted in a HisG variant that is still highly active even at L-histidine concentrations close to the solubility limit. Notably, the S143F mutation on its own provokes feedback deregulation, revealing for the first time an amino acid residue in the catalytic domain of HisG that is involved in the feedback regulatory mechanism. In addition, we investigated the effect of hisG mutations for L-histidine production on different levels. This comprised the analysis of different expression systems, including plasmid- and chromosome-based overexpression, as well as the importance of codon choice for HisG mutations. The combination of domain deletions, single amino acid exchanges, codon choice, and chromosome-based overexpression resulted in production strains accumulating around 0.5 g l(-1) L-histidine, demonstrating the added value of the different approaches. PMID:25892668

  9. Key enzymes of the protocatechuate branch of the β-ketoadipate pathway for aromatic degradation in Corynebacterium glutamicum

    Institute of Scientific and Technical Information of China (English)

    SHEN; Xihui; LIU; Shuangjiang

    2005-01-01

    Although the protocatechuate branch of the β-ketoadipate pathway in Gram bacteria has been well studied, this branch is less understood in Gram+ bacteria. In this study,Corynebacterium glutamicum was cultivated with protocatechuate, p-cresol, vanillate and 4-hydroxybenzoate as sole carbon and energy sources for growth. Enzymatic assays indicated that growing cells on these aromatic compounds exhibited protocatechuate 3,4-dioxygenase activities. Data-mining of the genome of this bacterium revealed that the genetic locus ncg12314-ncg12315 encoded a putative protocatechuate 3,4-dioxygenase. The genes,ncg12314 and ncg12315, were amplified by PCR technique and were cloned into plasmid (pET21aP34D). Recombinant Escherichia coli strain harboring this plasmid actively expressed protocatechuate 3,4-dioxygenase activity. Further, when this locus was disrupted in C. glutamicum, the ability to degrade and assimilate protocatechuate, p-cresol, vanillate or 4-hydroxybenzoate was lost and protocatechuate 3,4-dioxygenase activity was disappeared. The ability to grow with these aromatic compounds and protocatechuate 3,4-dioxygenase activity of C.glutamicum mutant could be restored by gene complementation. Thus, it is clear that the key enzyme for ring-cleavage, protocatechuate 3,4-dioxygenase, was encoded by ncg12314 and ncg12315. The additional genes involved in the protocatechuate branch of the β-ketoadipate pathway were identified by mining the genome data publically available in the GenBank. The functional identification of genes and their unique organization in C. glutamicum provided new insight into the genetic diversity of aromatic compound degradation.

  10. Improvement of the redox balance increases L-valine production by Corynebacterium glutamicum under oxygen deprivation conditions.

    Science.gov (United States)

    Hasegawa, Satoshi; Uematsu, Kimio; Natsuma, Yumi; Suda, Masako; Hiraga, Kazumi; Jojima, Toru; Inui, Masayuki; Yukawa, Hideaki

    2012-02-01

    Production of L-valine under oxygen deprivation conditions by Corynebacterium glutamicum lacking the lactate dehydrogenase gene ldhA and overexpressing the L-valine biosynthesis genes ilvBNCDE was repressed. This was attributed to imbalanced cofactor production and consumption in the overall L-valine synthesis pathway: two moles of NADH was generated and two moles of NADPH was consumed per mole of L-valine produced from one mole of glucose. In order to solve this cofactor imbalance, the coenzyme requirement for L-valine synthesis was converted from NADPH to NADH via modification of acetohydroxy acid isomeroreductase encoded by ilvC and introduction of Lysinibacillus sphaericus leucine dehydrogenase in place of endogenous transaminase B, encoded by ilvE. The intracellular NADH/NAD(+) ratio significantly decreased, and glucose consumption and L-valine production drastically improved. Moreover, L-valine yield increased and succinate formation decreased concomitantly with the decreased intracellular redox state. These observations suggest that the intracellular NADH/NAD(+) ratio, i.e., reoxidation of NADH, is the primary rate-limiting factor for L-valine production under oxygen deprivation conditions. The L-valine productivity and yield were even better and by-products derived from pyruvate further decreased as a result of a feedback resistance-inducing mutation in the acetohydroxy acid synthase encoded by ilvBN. The resultant strain produced 1,470 mM L-valine after 24 h with a yield of 0.63 mol mol of glucose(-1), and the L-valine productivity reached 1,940 mM after 48 h.

  11. Bioconversion of Gibberellin Fermentation Residue into Feed Supplement and Organic Fertilizer Employing Housefly (Musca domestica L. Assisted by Corynebacterium variabile.

    Directory of Open Access Journals (Sweden)

    Sen Yang

    Full Text Available The accumulation of a considerable quantity of gibberellin fermentation residue (GFR during gibberellic acid A3 (GA3 production not only results in the waste of many resources, but also poses a potential hazard to the environment, indicating that the safe treatment of GFR has become an urgent issue for GA3 industry. The key to recycle GFR is converting it into an available resource and removing the GA3 residue. To this end, we established a co-bioconversion process in this study using house fly larvae (HFL and microbes (Corynebacterium variabile to convert GFR into insect biomass and organic fertilizer. About 85.5% GA3 in the GFR was removed under the following optimized solid-state fermentation conditions: 60% GFR, 40% rice straw powder, pH 8.5 and 6 days at 26 °C. A total of 371 g housefly larvae meal and 2,064 g digested residue were bio-converted from 3,500 g raw GFR mixture contaning1, 400 g rice straw in the unit of (calculated dry matter. HFL meal derived from GFR contained 56.4% protein, 21.6% fat, and several essential amino acids, suggesting that it is a potential alternative animal feed protein source. Additionally, the digested GFR could be utilized as an organic fertilizer with a content of 3.2% total nitrogen, 2.0% inorganic phosphorus, 1.3% potassium and 91.5% organic matter. This novel GFR bio-conversion method can mitigate potential environmental pollution and recycle the waste resources.

  12. Bioconversion of Gibberellin Fermentation Residue into Feed Supplement and Organic Fertilizer Employing Housefly (Musca domestica L.) Assisted by Corynebacterium variabile.

    Science.gov (United States)

    Yang, Sen; Xie, Jiufeng; Hu, Nan; Liu, Yixiong; Zhang, Jiner; Ye, Xiaobin; Liu, Ziduo

    2015-01-01

    The accumulation of a considerable quantity of gibberellin fermentation residue (GFR) during gibberellic acid A3 (GA3) production not only results in the waste of many resources, but also poses a potential hazard to the environment, indicating that the safe treatment of GFR has become an urgent issue for GA3 industry. The key to recycle GFR is converting it into an available resource and removing the GA3 residue. To this end, we established a co-bioconversion process in this study using house fly larvae (HFL) and microbes (Corynebacterium variabile) to convert GFR into insect biomass and organic fertilizer. About 85.5% GA3 in the GFR was removed under the following optimized solid-state fermentation conditions: 60% GFR, 40% rice straw powder, pH 8.5 and 6 days at 26 °C. A total of 371 g housefly larvae meal and 2,064 g digested residue were bio-converted from 3,500 g raw GFR mixture contaning1, 400 g rice straw in the unit of (calculated) dry matter. HFL meal derived from GFR contained 56.4% protein, 21.6% fat, and several essential amino acids, suggesting that it is a potential alternative animal feed protein source. Additionally, the digested GFR could be utilized as an organic fertilizer with a content of 3.2% total nitrogen, 2.0% inorganic phosphorus, 1.3% potassium and 91.5% organic matter. This novel GFR bio-conversion method can mitigate potential environmental pollution and recycle the waste resources.

  13. Analysis of SOS-induced spontaneous prophage induction in Corynebacterium glutamicum at the single-cell level.

    Science.gov (United States)

    Nanda, Arun M; Heyer, Antonia; Krämer, Christina; Grünberger, Alexander; Kohlheyer, Dietrich; Frunzke, Julia

    2014-01-01

    The genome of the Gram-positive soil bacterium Corynebacterium glutamicum ATCC 13032 contains three integrated prophage elements (CGP1 to -3). Recently, it was shown that the large lysogenic prophage CGP3 (∼187 kbp) is excised spontaneously in a small number of cells. In this study, we provide evidence that a spontaneously induced SOS response is partly responsible for the observed spontaneous CGP3 induction. Whereas previous studies focused mainly on the induction of prophages at the population level, we analyzed the spontaneous CGP3 induction at the single-cell level using promoters of phage genes (Pint2 and Plysin) fused to reporter genes encoding fluorescent proteins. Flow-cytometric analysis revealed a spontaneous CGP3 activity in about 0.01 to 0.08% of the cells grown in standard minimal medium, which displayed a significantly reduced viability. A PrecA-eyfp promoter fusion revealed that a small fraction of C. glutamicum cells (∼0.2%) exhibited a spontaneous induction of the SOS response. Correlation of PrecA to the activity of downstream SOS genes (PdivS and PrecN) confirmed a bona fide induction of this stress response rather than stochastic gene expression. Interestingly, the reporter output of PrecA and CGP3 promoter fusions displayed a positive correlation at the single-cell level (ρ = 0.44 to 0.77). Furthermore, analysis of the PrecA-eyfp/Pint2-e2-crimson strain during growth revealed the highest percentage of spontaneous PrecA and Pint2 activity in the early exponential phase, when fast replication occurs. Based on these studies, we postulate that spontaneously occurring DNA damage induces the SOS response, which in turn triggers the induction of lysogenic prophages.

  14. Effect of biotin on transcription levels of key enzymes and glutamate efflux in glutamate fermentation by Corynebacterium glutamicum.

    Science.gov (United States)

    Cao, Yan; Duan, Zuoying; Shi, Zhongping

    2014-02-01

    Biotin is an important factor affecting the performance of glutamate fermentation by biotin auxotrophic Corynebacterium glutamicum and glutamate is over-produced only when initial biotin content is controlled at suitable levels or initial biotin is excessive but with Tween 40 addition during fermentation. The transcription levels of key enzymes at pyruvate, isocitrate and α-ketoglutarate metabolic nodes, as well as transport protein (TP) of glutamate were investigated under the conditions of varied biotin contents and Tween 40 supplementation. When biotin was insufficient, the genes encoding key enzymes and TP were down-regulated in the early production phase, in particular, the transcription level of isocitrate dehydrogenase (ICDH) which was only 2% of that of control. Although the cells' morphology transformation and TP level were not affected, low transcription level of ICDH led to lower final glutamate concentration (64 g/L). When biotin was excessive, the transcription levels of key enzymes were at comparable levels as those of control with ICDH as an exception, which was only 3-22% of control level throughout production phase. In this case, little intracellular glutamate accumulation (1.5 mg/g DCW) and impermeable membrane resulted in non glutamate secretion into broth, even though the quantity of TP was more than 10-folds of control level. Addition of Tween 40 when biotin was excessive stimulated the expression of all key enzymes and TP, intracellular glutamate content was much higher (10-12 mg/g DCW), and final glutamate concentration reached control level (75-80 g/L). Hence, the membrane alteration and TP were indispensable in glutamate secretion. Biotin and Tween 40 influenced the expression level of ICDH and glutamate efflux, thereby influencing glutamate production.

  15. Chemoimmunotherapy of small cell bronchogenic carcinoma with VP-16-213, ifosfamide, vincristine, adriamycin, and Corynebacterium parvum

    Energy Technology Data Exchange (ETDEWEB)

    Valdivieso, M.; Tenczynski, T.F.; Rodriguez, V.; Burgess, M.A.; Mountain, C.F.; Barkley, H.T. Jr.; Hersh, E.M.; Bodey, G.P.

    1981-07-15

    Thirty-five consecutive patients with small cell bronchogenic carcinoma (SCBC) received chemoimmunotherapy with VP-16-213, Ifosfamide, vincristine, Adriamycin, and Corynebacterium parvum. Of 33 evaluable patients, 26 (79%) responded with complete (55%) or partial (24%) remissions. Complete remissions were more common among patients with limited disease (11/14 patients, 79%) compared with those with extensive disease (7/19 patients, 37%) and among patients (11/14 patients, 79%) compared with those with extensive disease (7/19 patients, 37%) and among patients who were ambulatory prior to therapy (16/25 patients, 64%) compared with those who were nonambulatory (2/8 patients, 25%). Myelosuppression consisted primarily of neutropenia. Eight percent of the treatment courses in 29% of the patients were associated with hematuria and/or documented episodes of infection during neutropenia. There were three deaths possibly related to treatment, in two of which there was no evidence of disease at post-mortem examination. Six patients relapsed in the central nervous system (CNS). In four instances, CNS relapse was the only site of tumor progression. Central nervous system relapse was more common among evaluable patients who did not receive prophylactic brain irradiation (5/17 patients, 29%, vs. 1/15 patients, 7%; P . 0.23). The median survival duration for all patients was 63 weeks, being slightly longer for patients with limited disease than for those with extensive disease (70.9 weeks vs. 56 weeks; P . 0.18). This was also true for patients who achieved complete rather than partial remissions (71 weeks vs. 50 weeks; P . 0.09). Patients receiving prophylactic brain irradiation experienced longer survival (100.8 weeks vs. 48 weeks; P . 0.01).

  16. Regulation of the malic enzyme gene malE by the transcriptional regulator MalR in Corynebacterium glutamicum.

    Science.gov (United States)

    Krause, Jens P; Polen, Tino; Youn, Jung-Won; Emer, Denise; Eikmanns, Bernhard J; Wendisch, Volker F

    2012-06-15

    Corynebacterium glutamicum is a Gram-positive nonpathogenic bacterium that is used for the biotechnological production of amino acids. Here, we investigated the transcriptional control of the malE gene encoding malic enzyme (MalE) in C. glutamicum ATCC 13032, which is known to involve the nitrogen regulator AmtR. Gel shift experiments using purified regulators RamA and RamB revealed binding of these regulators to the malE promoter. In DNA-affinity purification experiments a hitherto uncharacterized transcriptional regulator belonging to the MarR family was found to bind to malE promoter DNA and was designated as MalR. C. glutamicum cells overexpressing malR showed reduced MalE activities in LB medium or in minimal media with acetate, glucose, pyruvate or citrate. Deletion of malR positively affected MalE activities during growth in LB medium and minimal media with pyruvate, glucose or the TCA cycle dicarboxylates l-malate, succinate and fumarate. Transcriptional fusion analysis revealed elevated malE promoter activity in the malR deletion mutant during growth in pyruvate minimal medium suggesting that MalR acts as a repressor of malE. Purified MalR bound malE promoter DNA in gel shift experiments. Two MalR binding sites were identified in the malE promoter by mutational analysis. Thus, MalR contributes to the complex transcriptional control of malE which also involves RamA, RamB and AmtR. PMID:22261175

  17. Assessment of heavy metal tolerance and hexavalent chromium reducing potential of Corynebacterium paurometabolum SKPD 1204 isolated from chromite mine seepage

    Directory of Open Access Journals (Sweden)

    Satarupa Dey

    2016-07-01

    Full Text Available Corynebacterium paurometabolum SKPD 1204 (MTCC 8730, a heavy metal tolerant and chromate reducing bacterium isolated from chromite mine seepage of Odisha, India has been evaluated for chromate reduction under batch culture. The isolate was found to tolerate metals like Co(II, Cu(II, Ni(II, Mn(II, Zn(II, Fe(III and Hg(II along with Cr(VI and was resistant to different antibiotics as evaluated by disc-diffusion method. The isolate, SKPD 1204 was found to reduce 62.5% of 2 mM Cr(VI in Vogel Bonner broth within 8 days of incubation. Chromate reduction capability of SKPD 1204 decreased with increase in Cr(VI concentration, but increased with increase in cell density and attained its maximum at 1010 cells/mL. Chromate reducing efficiency of SKPD 1204 was promoted in the presence of glycerol and glucose, while the highest reduction was recorded at pH 7.0 and 35 °C. The reduction process was inhibited by divalent cations Zn(II, Cd(II, Cu(II, and Ni(II, but not by Mn(II. Anions like nitrate, phosphate, sulphate and sulphite was found to be inhibitory to the process of Cr(VI reduction. Similarly, sodium fluoride, carbonyl cyanide m-chlorophenylhydrazone, sodium azide and N, N,-Di cyclohexyl carboiimide were inhibitory to chromate reduction, while 2,4-dinitrophenol appeared to be neither promotive nor inhibitory to the process.

  18. Identification of mannose uptake and catabolism genes in Corynebacterium glutamicum and genetic engineering for simultaneous utilization of mannose and glucose.

    Science.gov (United States)

    Sasaki, Miho; Teramoto, Haruhiko; Inui, Masayuki; Yukawa, Hideaki

    2011-03-01

    Here, focus is on Corynebacterium glutamicum mannose metabolic genes with the aim to improve this industrially important microorganism's ability to ferment mannose present in mixed sugar substrates. cgR_0857 encodes C. glutamicum's protein with 36% amino acid sequence identity to mannose 6-phosphate isomerase encoded by manA of Escherichia coli. Its deletion mutant did not grow on mannose and exhibited noticeably reduced growth on glucose as sole carbon sources. In effect, C. glutamicum manA is not only essential for growth on mannose but also important in glucose metabolism. A double deletion mutant of genes encoding glucose and fructose permeases (ptsG and ptsF, respectively) of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was not able to grow on mannose unlike the respective single deletion mutants with mannose utilization ability. A mutant deficient in ptsH, a general PTS gene, did not utilize mannose. These indicate that the glucose-PTS and fructose-PTS are responsible for mannose uptake in C. glutamicum. When cultured with a glucose and mannose mixture, mannose utilization of manA-overexpressing strain CRM1 was significantly higher than that of its wild-type counterpart, but with a strong preference for glucose. ptsF-overexpressing strain CRM2 co-utilized mannose and glucose, but at a total sugar consumption rate much lower than that of the wild-type strain and CRM1. Strain CRM3 overexpressing both manA and ptsF efficiently co-utilized mannose and glucose. Under oxygen-deprived conditions, high volumetric productivity of organic acids concomitant with the simultaneous consumption of the mixed sugars was achieved by the densely packed growth-arrested CRM3 cells.

  19. Prevalence of diphtheria and tetanus antibodies and circulation of Corynebacterium diphtheriae in São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    K.G. Divino-Goes

    2007-12-01

    Full Text Available The introduction of routine vaccination against tetanus and diphtheria in Brazil has decreased the incidence and changed the epidemiology of both diseases. We then investigated the prevalence of Corynebacterium diphtheriae carrier status and diphtheria and tetanus immunity in São Paulo, Brazil. From November 2001 to March 2003, 374 individuals were tested for the presence of C. diphtheriae in the naso-oropharynx and of serum diphtheria and tetanus antibodies. Participants were all healthy individuals without acute or chronic pathologies and they were stratified by age as follows: 0-12 months and 1-4, 5-9, 10-14, 15-24, 25-39, 40-59, and ³60 years. Antibodies were assessed using a double-antigen ELISA. C. diphtheriae species were identified by biochemical analysis and toxigenicity was assessed by the Elek test. For diphtheria, full protection (antibodies ³0.1 IU/mL was present in 84% of the individuals, 15% had basic protection (antibodies ³0.01 and <0.1 IU/mL and 1% were susceptible (antibodies <0.01 IU/mL. Full tetanus protection (antibodies ³0.1 IU/mL was present in 79% of the participants, 18% had basic protection (antibodies ³0.01 and <0.1 IU/mL and 3% were susceptible (antibodies <0.01 IU/mL. The geometric mean of diphtheria and tetanus antibodies reached the highest values at 5-9 years and decreased until the 40-59-year age range, increasing again in individuals over 60 years. Three participants (0.8% were carriers of C. diphtheriae, all non-toxigenic strains. The present results demonstrate the clear need of periodic booster for tetanus and diphtheria vaccine in adolescents and adults after primary immunization in childhood.

  20. Engineering of Corynebacterium glutamicum to utilize methyl acetate, a potential feedstock derived by carbonylation of methanol with CO.

    Science.gov (United States)

    Choo, Seungjung; Um, Youngsoon; Han, Sung Ok; Woo, Han Min

    2016-04-20

    The possibilities to utilize one-carbon substrates (C1) like CO, methane and methanol have been explored as a cheap alternative feedstock in the biotechnology. For the first time, methyl acetate (MeOAc), which can be formed from carbonylation of methanol with CO, was demonstrated to be an alternative carbon source for the cell growth of Corynebacterium glutamicum as a model microbial cell factory. To do so, a carboxyl esterase activity was necessary to hydrolyze MeOAc to methanol and acetate. Although the wild-type has an unknown esterase activity to MeOAc, the activity was not high enough to grow from 270mM MeOAc as sole carbon source, reaching OD600 of 5.28±0.2 in 32h. Based on the literatures studied for the esterase, we chose three esterases (MekB of Pseudomonas veronii MEK700, AcmB of Gordonia sp. Strain TY-5, and Est of Pyrobaculum calidifontis VA1) and cloned into the wild-type. As a result, the recombinant C. glutamicum expressing the highly active MekB esterase (28.6±0.77U/mg protein) showed complete degradation of MeOAc and utilization of acetate, resulting in OD600 of 16.5±0.02at 24h. In addition, the recombinant strain exhibited the rapid degradation of MeOAc to methanol and acetate in 2h under anaerobic condition. Therefore, MeOAc can be used as another C1-derived carbon source in the biotechnology. PMID:26970052

  1. 棒状杆菌临床分离株药物敏感性及同源性分析%Antibiotic susceptibility and homology analysis of Corynebacterium isolated from clinical specimens

    Institute of Scientific and Technical Information of China (English)

    谢爱香

    2012-01-01

    目的 了解棒状杆菌感染在临床中的耐药状况及同源性.方法 用肉汤微量细菌定量法检测抗菌药物的最低抑菌浓度(MIC),随机扩增多态性DNA(RAPD-PCR)并对棒状杆菌进行基因分型.结果 184株棒状杆菌对青霉素、头孢噻肟、红霉素、环丙沙星、四环素、克林霉素的耐药率均超过50%,对多西环素、亚胺培南、庆大霉素、利福平的耐药率在60%~80%之间,未发现对万古霉素和利奈唑胺耐药.18株纹带分离株可分成5型,其中A型占14株(77.8%).结论 棒状杆菌对多种抗菌药物的敏感性降低,纹带棒状杆菌存在院内传播的趋势.%Objective To investigate the drug susceptibility and homology of Corynebacterium isolated from clinical specimens . Methods Minimal inhibitory concentrations (MICs) of Corynebacterium to 12 antibiotics were tested with home-brew broth dilution microplates and genotypes of Corynebacterium were characterized by random amplified polymorphic DNA -polymerase chain reaction (RAPD-PC R) .Results The drug resistance of 184 strains of Corynebacterium to penicillin , cefotaxime, erythromycin, cip-rofloxacin, tetracycline and clindamyci were more than 50%, and to doxycycline, imipenem , gentamicin and rifampicin were about 60%-80% ,to vancomycin and linezolid were 0% . Eighteen strains of Corynebacterium stratum could be divided into 5 types , a-mong which 14(77 .8% ) strains belonged to A-type .Conclusion The sensitivity of Corynebacterium to various antibiotics might be decreasing and there could be a tendency of in-hospital spread of Corynebacterium stratum .

  2. Characterization of aspartate kinase and homoserine dehydrogenase from Corynebacterium glutamicum IWJ001 and systematic investigation of L-isoleucine biosynthesis.

    Science.gov (United States)

    Dong, Xunyan; Zhao, Yue; Zhao, Jianxun; Wang, Xiaoyuan

    2016-06-01

    Previously we have characterized a threonine dehydratase mutant TD(F383V) (encoded by ilvA1) and an acetohydroxy acid synthase mutant AHAS(P176S, D426E, L575W) (encoded by ilvBN1) in Corynebacterium glutamicum IWJ001, one of the best L-isoleucine producing strains. Here, we further characterized an aspartate kinase mutant AK(A279T) (encoded by lysC1) and a homoserine dehydrogenase mutant HD(G378S) (encoded by hom1) in IWJ001, and analyzed the consequences of all these mutant enzymes on amino acids production in the wild type background. In vitro enzyme tests confirmed that AK(A279T) is completely resistant to feed-back inhibition by L-threonine and L-lysine, and that HD(G378S) is partially resistant to L-threonine with the half maximal inhibitory concentration between 12 and 14 mM. In C. glutamicum ATCC13869, expressing lysC1 alone led to exclusive L-lysine accumulation, co-expressing hom1 and thrB1 with lysC1 shifted partial carbon flux from L-lysine (decreased by 50.1 %) to L-threonine (4.85 g/L) with minor L-isoleucine and no L-homoserine accumulation, further co-expressing ilvA1 completely depleted L-threonine and strongly shifted carbon flux from L-lysine (decreased by 83.0 %) to L-isoleucine (3.53 g/L). The results demonstrated the strongly feed-back resistant TD(F383V) might be the main driving force for L-isoleucine over-synthesis in this case, and the partially feed-back resistant HD(G378S) might prevent the accumulation of toxic intermediates. Information exploited from such mutation-bred production strain would be useful for metabolic engineering. PMID:27033538

  3. (L)-Valine production with minimization of by-products' synthesis in Corynebacterium glutamicum and Brevibacterium flavum.

    Science.gov (United States)

    Hou, Xiaohu; Chen, Xinde; Zhang, Yue; Qian, He; Zhang, Weiguo

    2012-12-01

    Corynebacterium glutamicum ATCC13032 and Brevibacterium flavum JV16 were engineered for L-valine production by over-expressing ilvEBN ( r ) C genes at 31 °C in 72 h fermentation. Different strategies were carried out to reduce the by-products' accumulation in L-valine fermentation and also to increase the availability of precursor for L-valine biosynthesis. The native promoter of ilvA of C. glutamicum was replaced with a weak promoter MPilvA (P-ilvAM1CG) to reduce the biosynthetic rate of L-isoleucine. Effect of different relative dissolved oxygen on L-valine production and by-products' formation was recorded, indicating that 15 % saturation may be the most appropriate relative dissolved oxygen for L-valine fermentation with almost no L-lactic acid and L-glutamate formed. To minimize L-alanine accumulation, alaT and/or avtA was inactivated in C. glutamicum and B. flavum, respectively. Compared to high concentration of L-alanine accumulated by alaT inactivated strains harboring ilvEBN ( r ) C genes, L-alanine concentration was reduced to 0.18 g/L by C. glutamicum ATCC13032MPilvA△avtA pDXW-8-ilvEBN ( r ) C, and 0.22 g/L by B. flavum JV16avtA::Cm pDXW-8-ilvEBN ( r ) C. Meanwhile, L-valine production and conversion efficiency were enhanced to 31.15 g/L and 0.173 g/g by C. glutamicum ATCC13032MPilvA△avtA pDXW-8-ilvEBN ( r ) C, 38.82 g/L and 0.252 g/g by B. flavum JV16avtA::Cm pDXW-8-ilvEBN ( r ) C. This study provides combined strategies to improve L-valine yield by minimization of by-products' production.

  4. Transcriptional response of Corynebacterium glutamicum ATCC 13032 to hydrogen peroxide stress and characterization of the OxyR regulon.

    Science.gov (United States)

    Milse, Johanna; Petri, Kathrin; Rückert, Christian; Kalinowski, Jörn

    2014-11-20

    The aerobic soil bacterium Corynebacterium glutamicum ATCC 13032 has a remarkable natural resistance to hydrogen peroxide. A major player in hydrogen peroxide defense is the LysR type transcriptional regulator OxyR, homologs of which are present in a wide range of bacteria. In this study, the global transcriptional response of C. glutamicum to oxidative stress induced by hydrogen peroxide was examined using whole genome DNA microarrays, demonstrating the dynamic reaction of the regulatory networks. Deletion of oxyR resulted in an increased resistance of the C. glutamicum mutant to hydrogen peroxide. By performing DNA microarray hybridizations and RT-qPCR, differentially expressed genes were detected in the mutant. The direct control by OxyR was verified by electrophoretic mobility shift assays for 12 target regions. The results demonstrated that OxyR in C. glutamicum acts as a transcriptional repressor under non-stress conditions for a total of 23 genes. The regulated genes encode proteins related to oxidative stress response (e.g. katA), iron homeostasis (e.g. dps) and sulfur metabolism (e.g. suf cluster). Besides the regulator of the suf cluster, SufR, OxyR regulated the gene cg1695 encoding a putative transcriptional regulator, indicating the role of OxyR as a master regulator in defense against oxidative stress. Using a modified DNase footprint approach, the OxyR-binding sites in five target promoter regions, katA, cydA, hemH, dps and cg1292, were localized and in each upstream region at least two overlapping binding sites were found. The DNA regions protected by the OxyR protein are about 56bp in length and do not have evident sequence similarities. Still, by giving an insight in the H2O2 stimulon and extending the OxyR regulon this study considerably contributes to the understanding of the response of C. glutamicum to hydrogen peroxide-mediated oxidative stress. PMID:25107507

  5. Molecular characterization of the Corynebacterium pseudotuberculosis hsp60-hsp10 operon, and evaluation of the immune response and protective efficacy induced by hsp60 DNA vaccination in mice

    Directory of Open Access Journals (Sweden)

    Oliveira Sérgio C

    2011-07-01

    Full Text Available Abstract Background Heat shock proteins (HSPs are important candidates for the development of vaccines because they are usually able to promote both humoral and cellular immune responses in mammals. We identified and characterized the hsp60-hsp10 bicistronic operon of the animal pathogen Corynebacterium pseudotuberculosis, a Gram-positive bacterium of the class Actinobacteria, which causes caseous lymphadenitis (CLA in small ruminants. Findings To construct the DNA vaccine, the hsp60 gene of C. pseudotuberculosis was cloned in a mammalian expression vector. BALB/c mice were immunized by intramuscular injection with the recombinant plasmid (pVAX1/hsp60. Conclusion This vaccination induced significant anti-hsp60 IgG, IgG1 and IgG2a isotype production. However, immunization with this DNA vaccine did not confer protective immunity.

  6. Study on the fermentation process conditions of L-valine produced by Corynebacterium glutamicum%L-缬氨酸的发酵工艺条件研究

    Institute of Scientific and Technical Information of China (English)

    曾青兰; 孙连连; 王志勇

    2012-01-01

    以谷氨酸棒杆菌(Corynebacterium glutamicum)CICC20887为生产菌株,采用单因素实验研究了发酵工艺条件对L-缬氨酸产量的影响.结果表明,该菌发酵生产L-缬氨酸的适宜初糖浓度、生物素添加量、VB1添加量、玉米浆添加量分别为90 g/L、80 μg/L、0.20 mg/L、30 g/L,发酵期间,24 h前pH值应控制在6.5~6.7、后48 h应控制在7.0~7.2,温度30~31℃,发酵周期应控制在66~72 h.%Using Corynebacterium glutamicum CICC20887 as producing strain,the effect of fermentation process condition on the yield of L-valine were studied with single factor design; The results showed that the optimum concentration of initial glucose, D-biotin, VB1 and corn syrup were 90g/L,80μg/L,0. 20 mg /L, 30g/L respectively;The optimal fermentation conditions of pH, temperature and fermentation period were 6. 5~6. 7 (for the initial 24 hours) ,7. 0~7. 2 (during 24 to 72 hours), 30~31℃ , 66~72 h respectively. This study could offe a basis for industrial production of L-valine.

  7. Quinone-dependent D-lactate dehydrogenase Dld (Cg1027 is essential for growth of Corynebacterium glutamicum on D-lactate

    Directory of Open Access Journals (Sweden)

    Oikawa Tadao

    2010-12-01

    Full Text Available Abstract Background Corynebacterium glutamicum is able to grow with lactate as sole or combined carbon and energy source. Quinone-dependent L-lactate dehydrogenase LldD is known to be essential for utilization of L-lactate by C. glutamicum. D-lactate also serves as sole carbon source for C. glutamicum ATCC 13032. Results Here, the gene cg1027 was shown to encode the quinone-dependent D-lactate dehydrogenase (Dld by enzymatic analysis of the protein purified from recombinant E. coli. The absorption spectrum of purified Dld indicated the presence of FAD as bound cofactor. Inactivation of dld resulted in the loss of the ability to grow with D-lactate, which could be restored by plasmid-borne expression of dld. Heterologous expression of dld from C. glutamicum ATCC 13032 in C. efficiens enabled this species to grow with D-lactate as sole carbon source. Homologs of dld of C. glutamicum ATCC 13032 are not encoded in the sequenced genomes of other corynebacteria and mycobacteria. However, the dld locus of C. glutamicum ATCC 13032 shares 2367 bp of 2372 bp identical nucleotides with the dld locus of Propionibacterium freudenreichii subsp. shermanii, a bacterium used in Swiss-type cheese making. Both loci are flanked by insertion sequences of the same family suggesting a possible event of horizontal gene transfer. Conclusions Cg1067 encodes quinone-dependent D-lactate dehydrogenase Dld of Corynebacterium glutamicum. Dld is essential for growth with D-lactate as sole carbon source. The genomic region of dld likely has been acquired by horizontal gene transfer.

  8. Scientific Opinion on the safety and efficacy of L-valine (ValAMINO®) produced by Corynebacterium glutamicum (DSM 25202) for all animal species, based on a dossier submitted by Evonik Industries AG

    OpenAIRE

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP)

    2014-01-01

    The product L-valine, feed grade (ValAMINO®), is a feed additive produced by fermentation with a genetically modified strain of Corynebacterium glutamicum (DSM 25202). Neither the production strain nor its recombinant DNA was detected in the final product. Therefore, the final product does not give rise to any safety concerns with regard to the genetic modification. The additive L-valine, feed grade, produced by C. glutamicum (DSM 25202) is safe for all target animals when supplemented in app...

  9. D-Allulose Production from D-Fructose by Permeabilized Recombinant Cells of Corynebacterium glutamicum Cells Expressing D-Allulose 3-Epimerase Flavonifractor plautii

    Science.gov (United States)

    Park, Chul-Soon; Kim, Taeyong; Hong, Seung-Hye; Shin, Kyung-Chul; Kim, Kyoung-Rok; Oh, Deok-Kun

    2016-01-01

    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 conditions, 10 g/L permeabilized cells produced 235 g/L d-allulose from 750 g/L d-fructose after 40 min, with a conversion rate of 31% (w/w) and volumetric productivity of 353 g/L/h, which were 1.4- and 2.1-fold higher than those obtained for nonpermeabilized cells, respectively. PMID:27467527

  10. Corynebacterium glutamicum ggtB encodes a functional γ-glutamyl transpeptidase with γ-glutamyl dipeptide synthetic and hydrolytic activity.

    Science.gov (United States)

    Walter, Frederik; Grenz, Sebastian; Ortseifen, Vera; Persicke, Marcus; Kalinowski, Jörn

    2016-08-20

    In this work the role of γ-glutamyl transpeptidase in the metabolism of γ-glutamyl dipeptides produced by Corynebacterium glutamicum ATCC 13032 was studied. The enzyme is encoded by the gene ggtB (cg1090) and synthesized as a 657 amino acids long preprotein. Gamma-glutamyl transpeptidase activity was found to be associated with intact cells of C. glutamicum and was abolished upon deletion of ggtB. Bioinformatic analysis indicated that the enzyme is a lipoprotein and is attached to the outer side of the cytoplasmic membrane. Biochemical parameters of recombinant GgtB were determined using the chromogenic substrate γ-glutamyl-p-nitroanilide. Highest activity of the enzyme was measured in sodium bicarbonate buffer at pH 9.6 and 45°C. The KM value was 123μM. GgtB catalyzed the concentration-dependent synthesis and hydrolysis of γ-glutamyl dipeptides and showed strong glutaminase activity. The intracellular concentrations of five γ-glutamyl dipeptides (γ-Glu-Glu, γ-Glu-Gln, γ-Glu-Val, γ-Glu-Leu, γ-Glu-Met) were determined by HPLC-MS and ranged from 0.15 to 0.4mg/g CDW after exponential growth in minimal media. Although deletion and overexpression of ggtB had significant effects on intracellular dipeptide concentrations, it was neither essential for biosynthesis nor catabolism of these dipeptides in vivo. PMID:26528625

  11. A thiol-disulfide oxidoreductase of the Gram-positive pathogen Corynebacterium diphtheriae is essential for viability, pilus assembly, toxin production and virulence

    Science.gov (United States)

    Reardon-Robinson, Melissa E.; Osipiuk, Jerzy; Jooya, Neda; Chang, Chungyu; Joachimiak, Andrzej; Das, Asis; Ton-That, Hung

    2016-01-01

    Summary The Gram-positive pathogen Corynebacterium diphtheriae exports through the Sec apparatus many extracellular proteins that include the key virulence factors diphtheria toxin and the adhesive pili. How these proteins attain their native conformations after translocation as unfolded precursors remains elusive. The fact that the majority of these exported proteins contain multiple cysteine residues and that several membrane-bound oxidoreductases are encoded in the corynebacterial genome suggests the existence of an oxidative protein-folding pathway in this organism. Here we show that the shaft pilin SpaA harbors a disulfide bond in vivo and alanine substitution of these cysteines abrogates SpaA polymerization and leads to the secretion of degraded SpaA peptides. We then identified a thiol-disulfide oxidoreductase (MdbA), whose structure exhibits a conserved thioredoxin-like domain with a CPHC active site. Remarkably, deletion of mdbA results in a severe temperature-sensitive cell division phenotype. This mutant also fails to assemble pilus structures and is greatly defective in toxin production. Consistent with these defects, the ΔmdbA mutant is attenuated in a guinea pig model of diphtheritic toxemia. Given its diverse cellular functions in cell division, pilus assembly and toxin production, we propose that MdbA is a component of the general oxidative folding machine in C. diphtheriae. PMID:26294390

  12. Roles of export genes cgmA and lysE for the production of L-arginine and L-citrulline by Corynebacterium glutamicum.

    Science.gov (United States)

    Lubitz, Dorit; Jorge, João M P; Pérez-García, Fernando; Taniguchi, Hironori; Wendisch, Volker F

    2016-10-01

    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 was shown to compensate for the lack of lysE with regard to L-arginine export. Moreover, plasmid-borne overexpression of cgmA rescued the toxic effect caused by feeding of the dipeptide Arg-Ala to lysE-deficient C. glutamicum and argO-deficient Escherichia coli strains. Deletion of the repressor gene cgmR improved L-arginine titers by 5 %. Production of L-lysine and L-citrulline was not affected by cgmA overexpression. Taken together, CgmA may function as an export system not only for the diamine putrescine and cadaverine but also for L-arginine. The major export system for L-lysine and L-arginine LysE may also play a role in L-citrulline export since production of L-citrulline was reduced when lysE was deleted and improved by 45 % when lysE was overproduced.

  13. Methionine uptake in Corynebacterium glutamicum by MetQNI and by MetPS, a novel methionine and alanine importer of the NSS neurotransmitter transporter family.

    Science.gov (United States)

    Trötschel, Christian; Follmann, Martin; Nettekoven, Jeannine A; Mohrbach, Tobias; Forrest, Lucy R; Burkovski, Andreas; Marin, Kay; Krämer, Reinhard

    2008-12-01

    The soil bacterium Corynebacterium glutamicum is a model organism in amino acid biotechnology. Here we present the identification of two different L-methionine uptake systems including the first characterization of a bacterial secondary methionine carrier. The primary carrier MetQNI is a high affinity ABC-type transporter specific for l-methionine. Its expression is under the control of the transcription factor McbR, the global regulator of sulfur metabolism in C. glutamicum. Besides MetQNI, a novel secondary methionine uptake system of the NSS (neurotransmitter:sodium symporter) family was identified and named MetP. The MetP system is characterized by a lower affinity for methionine and uses Na(+) ions for energetic coupling. It is also the main alanine transporter in C. glutamicum and is expressed constitutively. These observations are consistent with models of methionine, alanine, and leucine bound to MetP, derived from the X-ray crystal structure of the LeuT transporter from Aquifex aeolicus. Complementation studies show that MetP consists of two components, a large subunit with 12 predicted transmembrane segments and, surprisingly, an additional subunit with one predicted transmembrane segment only. Thus, this new member of the NSS transporter family adds a novel feature to this class of carriers, namely, the functional dependence on an additional small subunit.

  14. Evaluation of the food grade expression systems NICE and pSIP for the production of 2,5-diketo-D-gluconic acid reductase from Corynebacterium glutamicum.

    Science.gov (United States)

    Kaswurm, Vanja; Nguyen, Tien-Thanh; Maischberger, Thomas; Kulbe, Klaus D; Michlmayr, Herbert

    2013-01-28

    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 satisfies food safety requirements. In the present study, three recently described food grade variants of the Lactobacillales based expression systems pSIP (Lactobacillus plantarum) and NICE (Lactococcus lactis) were evaluated with regard to their effictiveness to produce 2,5-DKG reductase from Corynebacterium glutamicum. Our results indicate that both systems are suitable for 2,5-DKG reductase expression. Maximum production yields were obtained with Lb. plantarum/pSIP609 by pH control at 6.5. With 262 U per litre of broth, this represents the highest heterologous expression level so far reported for 2,5-DKG reductase from C. glutamicum. Accordingly, Lb. plantarum/pSIP609 might be an interesting alternative to Escherichia coli expression systems for industrial 2,5-DKG reductase production.

  15. B7-1/CD80-transduced tumor cells elicit better systemic immunity than wild-type tumor cells admixed with Corynebacterium parvum.

    Science.gov (United States)

    Chen, L; McGowan, P; Ashe, S; Johnston, J V; Hellström, I; Hellström, K E

    1994-10-15

    Tumor cells genetically modified by transduction of B7 (B7-1/CD80), a natural ligand for the T-cell costimulatory molecules CD28 and CTLA-4, can elicit potent tumor immunity, and they can be effective for treatment of established cancers in animal models. In this study, three tumor lines, the EL4 lymphoma, the P815 mastocytoma, and the MCA102 sarcoma were transduced with recombinant retrovirus containing the murine B7 gene, and their potency to induce systemic immunity protective against challenge with wild-type tumor was compared to that of the same tumor cells admixed with the commonly used adjuvant Corynebacterium parvum. While admixture of tumor cells with C. parvum resulted in complete regression of tumors in syngeneic mice, it did not induce protective immunity against a subsequent challenge of wild-type cells from any of the 3 tumors tested. In contrast, B7-transduced EL4 and P815 tumors regressed locally and induced a potent systemic immunity to wild-type tumors and a higher level of cytotoxic T-cell activity than did tumor cells admixed with C. parvum. No systemic immunity was induced by B7-transduced nonimmunogenic MCA102 sarcoma cells. Our results demonstrate that immunogenic tumor cells transduced with the B7 gene are superior to tumor cells mixed with C. parvum for the induction of systemic tumor immunity. PMID:7522958

  16. Metabolic flux distributions in Corynebacterium glutamicum during growth and lysine overproduction. Reprinted from Biotechnology and Bioengineering, Vol. 41, Pp 633-646 (1993).

    Science.gov (United States)

    Vallino, J J; Stephanopoulos, G

    2000-03-20

    The two main contributions of this article are the solidification of Corynebacterium glutamicum biochemistry guided by bioreaction network analysis, and the determination of basal metabolic flux distributions during growth and lysine synthesis. Employed methodology makes use of stoichiometrically based mass balances to determine flux distributions in the C. glutamicum metabolic network. Presented are a brief description of the methodology, a thorough literature review of glutamic acid bacteria biochemistry, and specific results obtained through a combination of fermentation studies and analysis-directed intracellular assays. The latter include the findings of the lack of activity of glyoxylate shunt, and that phosphoenolpyruvate carboxylase (PPC) is the only anaplerotic reaction expressed in C. glutamicum cultivated on glucose minimal media. Network simplifications afforded by the above findings facilitated the determination of metabolic flux distributions under a variety of culture conditions and led to the following conclusions. Both the pentose phosphate pathway and PPC support significant fluxes during growth and lysine overproduction, and that flux partitioning at the glucosa-6-phosphate branch point does not appear to limit lysine synthesis. PMID:10699864

  17. Isolation and structural characterization of Coryxin, a novel cyclic lipopeptide from Corynebacterium xerosis NS5 having emulsifying and anti-biofilm activity.

    Science.gov (United States)

    Dalili, Dina; Amini, Mohsen; Faramarzi, Mohammad Ali; Fazeli, Mohammad Reza; Khoshayand, Mohammad Reza; Samadi, Nasrin

    2015-11-01

    Herein we reported the structure and several properties of a new biosurfactants produced by Corynebacterium xerosis strain NS5. This strain was capable of producing a novel lipopeptide biosurfactant that we have named coryxin. The biosurfactant structure was characterized by using Fourier transform infrared spectroscopy (FTIR), Nuclear magnetic resonance spectroscopy (NMR), and Liquid chromatography-mass spectrometry (LC-MS). It contained a hydrophobic moiety of 3-hydroxydecanoic acid and a peptide part predicted as a sequence of seven amino acids including Asn-Arg-Asn-Gln-Pro-Asn-Ser. Coryxin lowered the surface tension of water to 31.4 mN/m, with a critical micelle concentration of 25mg/l. It was a strong emulsifier with an emulsification index of 61% against n-hexane. Coryxin showed antibacterial activity against test organisms belonging to Gram-positive and Gram-negative bacteria and disrupted preformed biofilms of Staphylococcus aureus (82.5%), Streptococcus mutans (80%), Escherichia coli (66%) and Pseudomonas aeruginosa (30%). In conclusion, microbial surfactant from C. xerosis exhibited inhibitory and disruptive activities against biofilm formation that could be of use in biofilm-related menace. PMID:26280817

  18. [Comparative electron-microscopic study of 8 representatives of the genus Corynebacterium grown on solid nutrient medium during the stationary phase of development].

    Science.gov (United States)

    Vysotskiĭ, V V; Mazurova, I K; Shmeleva, E A

    1976-09-01

    After 18 hours of growth on selective serum-agar medium C diphtheriae cultures with different toxicogenic activity, and also diphtheroid and Hoffmann's baccillus cultures were removed, washed of the remnants of the nutrient medium and fixed under cold conditions by two combined methods (with glutaric aldehyde-osmic acid--uranyl acetate, and potassium permeanganate--uranyl acetate). The preparations were studied in ultrathin sections. It appeared that corynebacteria had during the stationary phase of development a general structural plan characteristic of Gram positive microorganisms and for all the corynebacterium genus. Cells of diphtheria toxicognic strains had signs of the accelerated (in comparison with other strains) rate of development; the principal mass of toxicogenic cells after 18 hours of growth had morphological signs of the stage of rest. The majority of cells whose toxicogenicity was inconstant had an extensive microcapsule which was also a characteristic element of the diphtheroid and Hoffmann's bacillus ultrastructure. The total thickness of the walls in the cells of toxicogenic strains and of the strains whose toxicogenicity was inconstant constituted 190-200 A; in nontoxicogenic strains, diphtheroid and Hoffmann's bacillus it was from 230 to 320 A. Surface structures of corynebacteria were differentiated better in the cells with toxicogenic activity. In the majority of cells of nontoxicogenic strains and also diphtheroid and Hoffmann's bacillus individual wall layers were differentiated with difficulty. PMID:827882

  19. A de novo NADPH generation pathway for improving lysine production of Corynebacterium glutamicum by rational design of the coenzyme specificity of glyceraldehyde 3-phosphate dehydrogenase.

    Science.gov (United States)

    Bommareddy, Rajesh Reddy; Chen, Zhen; Rappert, Sugima; Zeng, An-Ping

    2014-09-01

    Engineering the cofactor availability is a common strategy of metabolic engineering to improve the production of many industrially important compounds. In this work, a de novo NADPH generation pathway is proposed by altering the coenzyme specificity of a native NAD-dependent glyceraldehyde 3-phosphate dehydrogenase (GAPDH) to NADP, which consequently has the potential to produce additional NADPH in the glycolytic pathway. Specifically, the coenzyme specificity of GAPDH of Corynebacterium glutamicum is systematically manipulated by rational protein design and the effect of the manipulation for cellular metabolism and lysine production is evaluated. By a combinatorial modification of four key residues within the coenzyme binding sites, different GAPDH mutants with varied coenzyme specificity were constructed. While increasing the catalytic efficiency of GAPDH towards NADP enhanced lysine production in all of the tested mutants, the most significant improvement of lysine production (~60%) was achieved with the mutant showing similar preference towards both NAD and NADP. Metabolic flux analysis with (13)C isotope studies confirmed that there was no significant change of flux towards the pentose phosphate pathway and the increased lysine yield was mainly attributed to the NADPH generated by the mutated GAPDH. The present study highlights the importance of protein engineering as a key strategy in de novo pathway design and overproduction of desired products.

  20. Development of an indirect ELISA to detect Corynebacterium pseudotuberculosis specific antibodies in sheep employing T1 strain culture supernatant as antigen

    Directory of Open Access Journals (Sweden)

    Miriam F. Rebouças

    2013-11-01

    Full Text Available Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis (CLA, a chronic disease that affects goats and sheep, characterized by granuloma formation in subcutaneous and internal lymph nodes. CLA causes significant economic losses to commercial goat herds. In this study, we aimed to test secreted antigens secreted from T1 strain bacteria grown in brain heart infusion (BHI broth in an indirect ELISA system to determine the presence of specific immunoglobulins against C. pseudotuberculosis. We analyzed the BHI antigen electrophoretic profile and the recognition pattern by infected sheep sera samples. The ELISA results were compared with multiplex PCR assay and IFN-gamma production. The ELISA was able to discriminate between negative and positive animals, with a sensitivity of 89% and a specificity of 99%, using microbiological isolation as gold standard. When this assay was compared with multiplex PCR and specific IFN-gamma quantification, six discrepant results were found among thirty-two samples. We concluded that the ELISA using antigens secreted from C. pseudotuberculosis T1 strain growth in BHI broth culture can be used for the serodiagnosis of CLA in sheep.

  1. Conservación por congelación de Bordetella pertussis y Corynebacterium diphtheriae, empleados en la producción de vacunas para uso humano

    Directory of Open Access Journals (Sweden)

    Yilian Plasencia,

    2000-11-01

    Full Text Available En el presente estudio se evaluó el método de congelación a –70ºC para la preservación de Bordetella pertussis y Corynebacterium diphtheriae. Para verificar el sustento de los cultivos se realizó un adecuado control de calidad, que incluyó comprobación de pureza, viabilidad y estabilidad de las propiedades de interés. En este trabajo se probaron diferentes formulaciones. Se seleccionó la que arrojó los mejores resultados y se realizó un estudio de mantenimiento de las características evaluadas durante el tiempo. Para medir determinados parámetros se realizaron procesos a escala industrial, empleándose para esto un biorreactor Chemap de 35 L. Se tomaron como referencia los valores obtenidos por las cepas conservadas por liofilización. De esta forma se buscaron alternativas y soluciones a problemas presentados en su conservación. Los resultados obtenidos sugieren la posible inclusión en el Programa de Mantenimiento establecido.

  2. The transcriptional regulatory network of Corynebacterium jeikeium K411 and its interaction with metabolic routes contributing to human body odor formation.

    Science.gov (United States)

    Barzantny, Helena; Schröder, Jasmin; Strotmeier, Jasmin; Fredrich, Eugenie; Brune, Iris; Tauch, Andreas

    2012-06-15

    Lipophilic corynebacteria are involved in the generation of volatile odorous products in the process of human body odor formation by degrading skin lipids and specific odor precursors. Therefore, these bacteria represent appropriate model systems for the cosmetic industry to examine axillary malodor formation on the molecular level. To understand the transcriptional control of metabolic pathways involved in this process, the transcriptional regulatory network of the lipophilic axilla isolate Corynebacterium jeikeium K411 was reconstructed from the complete genome sequence. This bioinformatic approach detected a gene-regulatory repertoire of 83 candidate proteins, including 56 DNA-binding transcriptional regulators, nine two-component systems, nine sigma factors, and nine regulators with diverse physiological functions. Furthermore, a cross-genome comparison among selected corynebacterial species of the taxonomic cluster 3 revealed a common gene-regulatory repertoire of 44 transcriptional regulators, including the MarR-like regulator Jk0257, which is exclusively encoded in the genomes of this taxonomical subline. The current network reconstruction comprises 48 transcriptional regulators and 674 gene-regulatory interactions that were assigned to five interconnected functional modules. Most genes involved in lipid degradation are under the combined control of the global cAMP-sensing transcriptional regulator GlxR and the LuxR-family regulator RamA, probably reflecting the essential role of lipid degradation in C. jeikeium. This study provides the first genome-scale in silico analysis of the transcriptional regulation of metabolism in a lipophilic bacterium involved in the formation of human body odor.

  3. Effect of biosurfactant and fertilizer on biodegradation of crude oil by marine isolates of Bacillus megaterium, Corynebacterium kutscheri and Pseudomonas aeruginosa.

    Science.gov (United States)

    Thavasi, Rengathavasi; Jayalakshmi, Singaram; Banat, Ibrahim M

    2011-01-01

    This study was conducted to investigate the effects of fertilizers and biosurfactants on biodegradation of crude oil by three marine bacterial isolates; Bacillus megaterium, Corynebacterium kutscheri and Pseudomonas aeruginosa. Five sets of experiments were carried out in shake flask and microcosm conditions with crude oil as follows: Set 1-only bacterial cells added (no fertilizer and biosurfactant), Set 2-with additional fertilizer only, Set 3-with additional biosurfactant only, Set 4-with added biosurfactant+fertilizer, Set 5-with no bacterial cells added (control), all the above experimental sets were incubated for 168 h. The biosurfactant+fertilizer added Set 4, resulted in maximum crude oil degradation within shake flask and microcosm conditions. Among the three bacterial isolates, P. aeruginosa and biosurfactant produced by this strain resulted in maximum crude oil degradation compared to the other two bacterial strains investigated. Interestingly, when biosurfactant and bacterial cells were used (Set 3), significant oil biodegradation activity occurred and the difference between this treatment and that in Set 4 with added fertilizer+biosurfactant were only 4-5% higher degradation level in shake flask and 3.2-7% in microcosm experiments for all three bacterial strains used. It is concluded that, biosurfactants alone capable of promoting biodegradation to a large extent without added fertilizers, which will reduce the cost of bioremediation process and minimizes the dilution or wash away problems encountered when water soluble fertilizers used during bioremediation of aquatic environments. PMID:20863694

  4. Parámetros fisicoquímicos para la síntesis de ácido láctico ó etanol de la bacteria (corynebacterium glutamicum)

    OpenAIRE

    Garcia Lina Marcela; Castellanos Sánchez Angélica María; Lopez Galan Jorge Enrique; Astudillo Myriam; Florez Pardo Luz Marina

    2011-01-01

    El interés por obtener productos para la industria de biocombustibles a partir de desechos agrícolas, conduce a las investigaciones en la búsqueda de sistemas microbianos resistentes y costo-efectivos. La Corynebacterium glutamicum, es un microorganismo usado para producir amino-ácidos, crece en gran variedad de sustratos y es resistente durante la fermentación, a variaciones en el pH, temperatura, presión osmótica y acumulación de alcohol, características que lo hacen candidato a ser mejora...

  5. Integration of E.coli aroG-pheA tandem genes into Corynebacterium glutamicum tyrA locus and its effect on L-phenylalanine biosynthesis

    Institute of Scientific and Technical Information of China (English)

    Dong-Xin Liu; Chang-Sheng Fan; Ju-Hong Tao; Guo-Xin Liang; Shan-E Gao; Hai-Jiao Wang; Xin Li; Da-Xin Song

    2004-01-01

    AIM: To study the effect of integration of tandem aroG-pheA genes into the tyrA locus of Corynebacterium glutamicum (C. glutamicum) on the production of L-phenylalanine.METHODS: By nitrosoguanidine mutagenesis, five pfluorophenylalanine (FP)-resistant mutants of C.glutamicurm FP were selected. The tyrA gene encoding prephenate dehydrogenase (PDH) of C.glutamicum was amplified by polymerase chain reaction (PCR) and cloned on the plasmid pPR. Kanamycin resistance gene (Km) and the PBF-aroGpheA-T (GA) fragment of pGA were inserted into tyrA gene to form targeting vectors pTK and pTGAK, respectively. Then,they were transformed into C.glutamicum FP respectively by electroporation. Cultures were screened by a medium containing kanamycin and detected by PCR and phenotype analysis. The transformed strains were used for L-phenylalanine fermentation and enzyme assays.RESULTS: Engineering strains of C.glutamicum (Tyr-) were obtained. Compared with the original strain, the transformed strain C. glutamicum GAK was observed to have the highest elevation of L-phenylalanine production by a 1.71-fold,and 2.9-, 3.36-, and 3.0-fold in enzyme activities of chorismate mutase, prephenate dehydratase and 3-deoxy-Darabinoheptulosonate-7-phosphate synthase, respectively.CONCLUSION: Integration of tandem aroG-pheA genes into tyrA locus of C. glutamicum chromosome can disrupt tyrA gene and increase the yield of L-phenylalanine production.

  6. Competition of Reactive red 4, Reactive orange 16 and Basic blue 3 during biosorption of Reactive blue 4 by polysulfone-immobilized Corynebacterium glutamicum

    International Nuclear Information System (INIS)

    Competition of Reactive red 4 (RR4), Reactive orange 16 (RO16) and Basic blue 3 (BB3) during biosorption of Reactive blue 4 (RB4) by polysulfone-immobilized protonated Corynebacterium glutamicum (PIPC) was investigated in batch and column mode of operations. Through potentiometric titrations, and with the aid of proton-binding model, carboxyl, phosphonate and amine were identified as functional groups of PIPC, with apparent pKa values of 3.47 ± 0.05, 7.08 ± 0.07 and 9.90 ± 0.05 mmol/g, respectively. Since reactive dyes release dye anions (ROSO3-) in solutions, the positively charged amine groups were responsible for biosorption. PIPC favored biosorption at pH 3 when RB4 was studied/used as single-solute; while the presence of RR4 and RO16 severely affected the RB4 biosorption. When present as a single-solute, PIPC recorded 184.5 mg RB4/g; while PIPC exhibited 126.9, 120.9 and 169.6 mg RB4/g in the presence of RR4, RO16 and BB3, respectively. In general, the accessibility of amine group depends on the molecular size, number of sulfonate groups and reactivity of each reactive dye. Single and multicomponent Freundlich equations successfully described the biosorption isotherms. With 0.1 M NaOH, it is possible to reuse PIPC for RB4 biosorption in 10 repeated cycles. Column experiments in an up-flow packed column coincided with batch results, that is PIPC showed strong preference towards highly reactive and relatively small RB4 anions; however, the presence of competing dyes hinder the RB4 column biosorption performance

  7. Application of a genetically encoded biosensor for live cell imaging of L-valine production in pyruvate dehydrogenase complex-deficient Corynebacterium glutamicum strains.

    Science.gov (United States)

    Mustafi, Nurije; Grünberger, Alexander; Mahr, Regina; Helfrich, Stefan; Nöh, Katharina; Blombach, Bastian; Kohlheyer, Dietrich; Frunzke, Julia

    2014-01-01

    The majority of biotechnologically relevant metabolites do not impart a conspicuous phenotype to the producing cell. Consequently, the analysis of microbial metabolite production is still dominated by bulk techniques, which may obscure significant variation at the single-cell level. In this study, we have applied the recently developed Lrp-biosensor for monitoring of amino acid production in single cells of gradually engineered L-valine producing Corynebacterium glutamicum strains based on the pyruvate dehydrogenase complex-deficient (PDHC) strain C. glutamicum ΔaceE. Online monitoring of the sensor output (eYFP fluorescence) during batch cultivation proved the sensor's suitability for visualizing different production levels. In the following, we conducted live cell imaging studies on C. glutamicum sensor strains using microfluidic chip devices. As expected, the sensor output was higher in microcolonies of high-yield producers in comparison to the basic strain C. glutamicum ΔaceE. Microfluidic cultivation in minimal medium revealed a typical Gaussian distribution of single cell fluorescence during the production phase. Remarkably, low amounts of complex nutrients completely changed the observed phenotypic pattern of all strains, resulting in a phenotypic split of the population. Whereas some cells stopped growing and initiated L-valine production, others continued to grow or showed a delayed transition to production. Depending on the cultivation conditions, a considerable fraction of non-fluorescent cells was observed, suggesting a loss of metabolic activity. These studies demonstrate that genetically encoded biosensors are a valuable tool for monitoring single cell productivity and to study the phenotypic pattern of microbial production strains.

  8. A slow-forming isopeptide bond in the structure of the major pilin SpaD from Corynebacterium diphtheriae has implications for pilus assembly

    International Nuclear Information System (INIS)

    Two crystal structures of the major pilin SpaD from C. diphtheriae have been determined at 1.87 and 2.5 Å resolution. The N-terminal domain is found to contain an isopeptide bond that forms slowly over time in the recombinant protein. Given its structural context, this provides insight into the relationship between internal isopeptide-bond formation and pilus assembly. The Gram-positive organism Corynebacterium diphtheriae, the cause of diphtheria in humans, expresses pili on its surface which it uses for adhesion and colonization of its host. These pili are covalent protein polymers composed of three types of pilin subunit that are assembled by specific sortase enzymes. A structural analysis of the major pilin SpaD, which forms the polymeric backbone of one of the three types of pilus expressed by C. diphtheriae, is reported. Mass-spectral and crystallographic analysis shows that SpaD contains three internal Lys–Asn isopeptide bonds. One of these, shown by mass spectrometry to be located in the N-terminal D1 domain of the protein, only forms slowly, implying an energy barrier to bond formation. Two crystal structures, of the full-length three-domain protein at 2.5 Å resolution and of a two-domain (D2-D3) construct at 1.87 Å resolution, show that each of the three Ig-like domains contains a single Lys–Asn isopeptide-bond cross-link, assumed to give mechanical stability as in other such pili. Additional stabilizing features include a disulfide bond in the D3 domain and a calcium-binding loop in D2. The N-terminal D1 domain is more flexible than the others and, by analogy with other major pilins of this type, the slow formation of its isopeptide bond can be attributed to its location adjacent to the lysine used in sortase-mediated polymerization during pilus assembly

  9. Administration of interleukin-10 at the time of priming protects Corynebacterium parvum-primed mice against LPS- and TNF-alpha-induced lethality.

    Science.gov (United States)

    Smith, S R; Terminelli, C; Denhardt, G; Narula, S; Thorbecke, G J

    1996-11-01

    Several laboratories have described the protective effects of interleukin-10 (IL-10) in mouse models of lethal endotoxemia. In most of these experiments, protection was observed in normal mice that were given a lethal dose of LPS. However, we failed to observe protection with IL-10 in LPS-challenged mice that had been primed with Corynebacterium parvum (Proprionibacterium acnes). We have extended our studies with IL-10 in C. parvum-primed mice and in some cases have observed protection that appears to depend on the strength of the sensitization to C. parvum. When IL-10 was administered to mice at the time of priming, it was particularly effective in blocking sensitization, as evidenced by the inability of treated mice to mount a strong inflammatory cytokine response when subsequently challenged with LPS. Following such treatment with IL-10, C. parvum-primed mice were also protected from a subsequent lethal challenge with rMuTNF-alpha. In addition, the mice were protected against LPS- and TNF-alpha-induced lethality with a single dose of an anti-TNF-alpha or anti-IFN-gamma mAb given at the time of priming. Our results suggest that TNF-alpha and IFN-gamma are produced early after priming with C. parvum and are at least partly responsible for the enhanced sensitivity of the mice to LPS and TNF-alpha. IL-10 affords protection to the mice because of its ability to block the C. parvum-induced TNF-alpha and IFN-gamma responses. PMID:8912878

  10. Single-Domain Peptidyl-Prolyl cis/trans Isomerase FkpA from Corynebacterium glutamicum Improves the Biomass Yield at Increased Growth Temperatures.

    Science.gov (United States)

    Kallscheuer, Nicolai; Bott, Michael; van Ooyen, Jan; Polen, Tino

    2015-11-01

    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 predicted single-domain FK-506 (tacrolimus) binding protein (FKBP)-type PPIase (FkpA) is encoded directly downstream of gltA, which encodes citrate synthase (CS). This gene cluster is also present in other Actinobacteria. Here we carried out in vitro and in vivo experiments to study the function and influence of predicted FkpA in C. glutamicum. In vitro, FkpA indeed shows typical PPIase activity with artificial substrates and is inhibited by FK-506. Furthermore, FkpA delays the aggregation of CS, which is also inhibited by FK-506. Surprisingly, FkpA has a positive effect on the activity and temperature range of CS in vitro. Deletion of fkpA causes a 50% reduced biomass yield compared to that of the wild type when grown at 37°C, whereas there is only a 10% reduced biomass yield at the optimal growth temperature of 30°C accompanied by accumulation of 7 mM l-glutamate and 22 mM 2-oxoglutarate. Thus, FkpA may be exploited for improved product formation in biotechnical processes. Comparative transcriptome analysis revealed 69 genes which exhibit ≥2-fold mRNA level changes in C. glutamicum ΔfkpA, giving insight into the transcriptional response upon mild heat stress when FkpA is absent. PMID:26341203

  11. Carbon flux analysis by 13C nuclear magnetic resonance to determine the effect of CO2 on anaerobic succinate production by Corynebacterium glutamicum.

    Science.gov (United States)

    Radoš, Dušica; Turner, David L; Fonseca, Luís L; Carvalho, Ana Lúcia; Blombach, Bastian; Eikmanns, Bernhard J; Neves, Ana Rute; Santos, Helena

    2014-05-01

    Wild-type Corynebacterium glutamicum produces a mixture of lactic, succinic, and acetic acids from glucose under oxygen deprivation. We investigated the effect of CO2 on the production of organic acids in a two-stage process: cells were grown aerobically in glucose, and subsequently, organic acid production by nongrowing cells was studied under anaerobic conditions. The presence of CO2 caused up to a 3-fold increase in the succinate yield (1 mol per mol of glucose) and about 2-fold increase in acetate, both at the expense of l-lactate production; moreover, dihydroxyacetone formation was abolished. The redistribution of carbon fluxes in response to CO2 was estimated by using (13)C-labeled glucose and (13)C nuclear magnetic resonance (NMR) analysis of the labeling patterns in end products. The flux analysis showed that 97% of succinate was produced via the reductive part of the tricarboxylic acid cycle, with the low activity of the oxidative branch being sufficient to provide the reducing equivalents needed for the redox balance. The flux via the pentose phosphate pathway was low (~5%) regardless of the presence or absence of CO2. Moreover, there was significant channeling of carbon to storage compounds (glycogen and trehalose) and concomitant catabolism of these reserves. The intracellular and extracellular pools of lactate and succinate were measured by in vivo NMR, and the stoichiometry (H(+):organic acid) of the respective exporters was calculated. This study shows that it is feasible to take advantage of natural cellular regulation mechanisms to obtain high yields of succinate with C. glutamicum without genetic manipulation. PMID:24610842

  12. A slow-forming isopeptide bond in the structure of the major pilin SpaD from Corynebacterium diphtheriae has implications for pilus assembly

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Hae Joo; Paterson, Neil G. [University of Auckland, Private Bag 92019, Auckland 1142 (New Zealand); Kim, Chae Un [Cornell University, Ithaca, NY 14853 (United States); Middleditch, Martin [University of Auckland, Private Bag 92019, Auckland 1142 (New Zealand); Chang, Chungyu; Ton-That, Hung [University of Texas–Houston Medical School, Houston, TX 77030 (United States); Baker, Edward N., E-mail: ted.baker@auckland.ac.nz [University of Auckland, Private Bag 92019, Auckland 1142 (New Zealand)

    2014-05-01

    Two crystal structures of the major pilin SpaD from C. diphtheriae have been determined at 1.87 and 2.5 Å resolution. The N-terminal domain is found to contain an isopeptide bond that forms slowly over time in the recombinant protein. Given its structural context, this provides insight into the relationship between internal isopeptide-bond formation and pilus assembly. The Gram-positive organism Corynebacterium diphtheriae, the cause of diphtheria in humans, expresses pili on its surface which it uses for adhesion and colonization of its host. These pili are covalent protein polymers composed of three types of pilin subunit that are assembled by specific sortase enzymes. A structural analysis of the major pilin SpaD, which forms the polymeric backbone of one of the three types of pilus expressed by C. diphtheriae, is reported. Mass-spectral and crystallographic analysis shows that SpaD contains three internal Lys–Asn isopeptide bonds. One of these, shown by mass spectrometry to be located in the N-terminal D1 domain of the protein, only forms slowly, implying an energy barrier to bond formation. Two crystal structures, of the full-length three-domain protein at 2.5 Å resolution and of a two-domain (D2-D3) construct at 1.87 Å resolution, show that each of the three Ig-like domains contains a single Lys–Asn isopeptide-bond cross-link, assumed to give mechanical stability as in other such pili. Additional stabilizing features include a disulfide bond in the D3 domain and a calcium-binding loop in D2. The N-terminal D1 domain is more flexible than the others and, by analogy with other major pilins of this type, the slow formation of its isopeptide bond can be attributed to its location adjacent to the lysine used in sortase-mediated polymerization during pilus assembly.

  13. Transcriptome sequencing revealed the transcriptional organization at ribosome-mediated attenuation sites in Corynebacterium glutamicum and identified a novel attenuator involved in aromatic amino acid biosynthesis.

    Science.gov (United States)

    Neshat, Armin; Mentz, Almut; Rückert, Christian; Kalinowski, Jörn

    2014-11-20

    The Gram-positive bacterium Corynebacterium glutamicum belongs to the order Corynebacteriales and is used as a producer of amino acids at industrial scales. Due to its economic importance, gene expression and particularly the regulation of amino acid biosynthesis has been investigated extensively. Applying the high-resolution technique of transcriptome sequencing (RNA-seq), recently a vast amount of data has been generated that was used to comprehensively analyze the C. glutamicum transcriptome. By analyzing RNA-seq data from a small RNA cDNA library of C. glutamicum, short transcripts in the known transcriptional attenuators sites of the trp operon, the ilvBNC operon and the leuA gene were verified. Furthermore, whole transcriptome RNA-seq data were used to elucidate the transcriptional organization of these three amino acid biosynthesis operons. In addition, we discovered and analyzed the novel attenuator aroR, located upstream of the aroF gene (cg1129). The DAHP synthase encoded by aroF catalyzes the first step in aromatic amino acid synthesis. The AroR leader peptide contains the amino acid sequence motif F-Y-F, indicating a regulatory effect by phenylalanine and tyrosine. Analysis by real-time RT-PCR suggests that the attenuator regulates the transcription of aroF in dependence of the cellular amount of tRNA loaded with phenylalanine when comparing a phenylalanine-auxotrophic C. glutamicum mutant fed with limiting and excess amounts of a phenylalanine-containing dipeptide. Additionally, the very interesting finding was made that all analyzed attenuators are leaderless transcripts. PMID:24910972

  14. 酶法处理短棒状杆菌的效果分析%Analysis of effect of enzyme-digested Corynebacterium parvum product

    Institute of Scientific and Technical Information of China (English)

    曹芳; 许弟群; 罗开梅; 郭立君

    2012-01-01

    Objective To investigate the effect of enzyme-digested Corynebacterium parvum product (ECCP). Methods ECPP was prepared by heating inactivation,ultrasonication and Pronase digestion,observed for structure by electron microscopy,and determined for protein,polysaccharide and nucleic acid contents. Pure BALB/c mice,Chinchilla rabbits and NIH mice were randomly divided into negative control (CK,0. 9% sodium chloride),C. parvum product (CPP,1 000 μg/ml) as well as ECPP I (ECPP 500 μg/ml) and II (ECPP 1 000 μg/ml) groups separately. The pure BALB/c mice were determined for spleen index,abnormal toxicily and serum IL-2,IFNγ and TNF-α levels,while Chinchilla rabbits for pyrogen reaction,and NIH mice for tumor-inhibiting effect. Results Compared with those of heat-inaclivated CP,the cell walls of ECPP were thin,while the intracellukr structure showed no significant change. The protein,polysaccharide and nucleic acid contents of ECPP decreased significantly as compared with those of CPP (P< 0. 01). The spleen index in ECPP II group was 2. 96,and the inhibitory effect on Ehrlich ascites tumor was observed,with a survival rate of animals of 80%. The rabbits in ECPP II group were qualified in pyrogen test. Abnormal toxicity test showed that the bodyweights of mice in ECPP II group increased significantly as compared with those in CPP group. However,the serum IL-2,IFNγ and TNF-α levels of mice in ECPP II group increased significantly. Conclusion ECPP decreased the adverse reaction and enhanced the antitumor function,which showed good antitumor effect and might be used as a novel immunomodulator of good prospect of application.%目的 探讨酶法处理短棒状杆菌(Enzyme-digested Corynebacterium parvum product,ECPP)的效果.方法 采用热灭活、超声、Pronase酶消化等方法处理短棒状杆菌(Corynebacterium parvum,CP),电镜观察其结构,并测定其蛋白、多糖及核酸含量.将BALB/c纯系小鼠、青紫蓝家兔和NIH小鼠均

  15. Platform engineering of Corynebacterium glutamicum with reduced pyruvate dehydrogenase complex activity for improved production of L-lysine, L-valine, and 2-ketoisovalerate.

    Science.gov (United States)

    Buchholz, Jens; Schwentner, Andreas; Brunnenkan, Britta; Gabris, Christina; Grimm, Simon; Gerstmeir, Robert; Takors, Ralf; Eikmanns, Bernhard J; Blombach, Bastian

    2013-09-01

    Exchange of the native Corynebacterium glutamicum promoter of the aceE gene, encoding the E1p subunit of the pyruvate dehydrogenase complex (PDHC), with mutated dapA promoter variants led to a series of C. glutamicum strains with gradually reduced growth rates and PDHC activities. Upon overexpression of the l-valine biosynthetic genes ilvBNCE, all strains produced l-valine. Among these strains, C. glutamicum aceE A16 (pJC4 ilvBNCE) showed the highest biomass and product yields, and thus it was further improved by additional deletion of the pqo and ppc genes, encoding pyruvate:quinone oxidoreductase and phosphoenolpyruvate carboxylase, respectively. In fed-batch fermentations at high cell densities, C. glutamicum aceE A16 Δpqo Δppc (pJC4 ilvBNCE) produced up to 738 mM (i.e., 86.5 g/liter) l-valine with an overall yield (YP/S) of 0.36 mol per mol of glucose and a volumetric productivity (QP) of 13.6 mM per h [1.6 g/(liter × h)]. Additional inactivation of the transaminase B gene (ilvE) and overexpression of ilvBNCD instead of ilvBNCE transformed the l-valine-producing strain into a 2-ketoisovalerate producer, excreting up to 303 mM (35 g/liter) 2-ketoisovalerate with a YP/S of 0.24 mol per mol of glucose and a QP of 6.9 mM per h [0.8 g/(liter × h)]. The replacement of the aceE promoter by the dapA-A16 promoter in the two C. glutamicum l-lysine producers DM1800 and DM1933 improved the production by 100% and 44%, respectively. These results demonstrate that C. glutamicum strains with reduced PDHC activity are an excellent platform for the production of pyruvate-derived products.

  16. The DtxR protein acting as dual transcriptional regulator directs a global regulatory network involved in iron metabolism of Corynebacterium glutamicum

    Directory of Open Access Journals (Sweden)

    Hüser Andrea T

    2006-02-01

    Full Text Available Abstract Background The knowledge about complete bacterial genome sequences opens the way to reconstruct the qualitative topology and global connectivity of transcriptional regulatory networks. Since iron is essential for a variety of cellular processes but also poses problems in biological systems due to its high toxicity, bacteria have evolved complex transcriptional regulatory networks to achieve an effective iron homeostasis. Here, we apply a combination of transcriptomics, bioinformatics, in vitro assays, and comparative genomics to decipher the regulatory network of the iron-dependent transcriptional regulator DtxR of Corynebacterium glutamicum. Results A deletion of the dtxR gene of C. glutamicum ATCC 13032 led to the mutant strain C. glutamicum IB2103 that was able to grow in minimal medium only under low-iron conditions. By performing genome-wide DNA microarray hybridizations, differentially expressed genes involved in iron metabolism of C. glutamicum were detected in the dtxR mutant. Bioinformatics analysis of the genome sequence identified a common 19-bp motif within the upstream region of 31 genes, whose differential expression in C. glutamicum IB2103 was verified by real-time reverse transcription PCR. Binding of a His-tagged DtxR protein to oligonucleotides containing the 19-bp motifs was demonstrated in vitro by DNA band shift assays. At least 64 genes encoding a variety of physiological functions in iron transport and utilization, in central carbohydrate metabolism and in transcriptional regulation are controlled directly by the DtxR protein. A comparison with the bioinformatically predicted networks of C. efficiens, C. diphtheriae and C. jeikeium identified evolutionary conserved elements of the DtxR network. Conclusion This work adds considerably to our currrent understanding of the transcriptional regulatory network of C. glutamicum genes that are controlled by DtxR. The DtxR protein has a major role in controlling the

  17. PARÁMETROS FISICOQUÍMICOS PARA LA SÍNTESIS DE ÁCIDO LÁCTICO Ó ETANOL DE LA BACTERIA (Corynebacterium glutamicum

    Directory of Open Access Journals (Sweden)

    Garcia Lina Marcela

    2011-08-01

    Full Text Available El interés por obtener productos para la industria de biocombustibles a partir de desechos agrícolas, conduce a las investigaciones en la búsqueda de sistemas microbianos resistentes y costo-efectivos. La Corynebacterium glutamicum, es un microorganismo usado para producir amino-ácidos, crece en gran variedad de sustratos y es resistente durante la fermentación, a variaciones en el pH, temperatura, presión osmótica y acumulación de alcohol, características que lo hacen candidato a ser mejorado para la síntesis de ácido láctico y etanol. Aún se desconocen aspectos de su fisiología que aumenten su eficiencia en convertir azúcares (C5 y C6 en estos dos metabolitos. Por tanto, este trabajo se basó en estudiar e identificar los parámetros fisicoquímicos que tuvieron un mayor efecto sobre el crecimiento bacteriano y la síntesis de ácido láctico. Para lograr este objetivo, ocho variables fueron evaluadas en un modelo estadístico producido en erlenmeyer, con estos resultados se hallaron las condiciones óptimas que fueron evaluadas en un cultivo por lotes en biorreactor. La temperatura, la concentración de biotina y azúcar fueron las variables con mayor impacto (p< 0,05 sobre el cultivo. Usando las condiciones óptimas, 36 °C; 6,1 mg/L de biotina y 50 g/L de glucosa, se obtuvo una max de 0,394 h-1, 16 g/L de ácido láctico a las 15 h del proceso con un rendimiento del 32%; observándose un mayor consumo de sustrato durante el crecimiento y poca disponibilidad para la fermentación, que sugiriendo una alimentación del cultivo al final de la fase exponencial que aumente los rendimientos de producción.

  18. Global gene expression during stringent response in Corynebacterium glutamicum in presence and absence of the rel gene encoding (pppGpp synthase

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    Kalinowski Jörn

    2006-09-01

    Full Text Available Background The stringent response is the initial reaction of microorganisms to nutritional stress. During stringent response the small nucleotides (pppGpp act as global regulators and reprogram bacterial transcription. In this work, the genetic network controlled by the stringent response was characterized in the amino acid-producing Corynebacterium glutamicum. Results The transcriptome of a C. glutamicum rel gene deletion mutant, unable to synthesize (pppGpp and to induce the stringent response, was compared with that of its rel-proficient parent strain by microarray analysis. A total of 357 genes were found to be transcribed differentially in the rel-deficient mutant strain. In a second experiment, the stringent response was induced by addition of DL-serine hydroxamate (SHX in early exponential growth phase. The time point of the maximal effect on transcription was determined by real-time RT-PCR using the histidine and serine biosynthetic genes. Transcription of all of these genes reached a maximum at 10 minutes after SHX addition. Microarray experiments were performed comparing the transcriptomes of SHX-induced cultures of the rel-proficient strain and the rel mutant. The differentially expressed genes were grouped into three classes. Class A comprises genes which are differentially regulated only in the presence of an intact rel gene. This class includes the non-essential sigma factor gene sigB which was upregulated and a large number of genes involved in nitrogen metabolism which were downregulated. Class B comprises genes which were differentially regulated in response to SHX in both strains, independent of the rel gene. A large number of genes encoding ribosomal proteins fall into this class, all being downregulated. Class C comprises genes which were differentially regulated in response to SHX only in the rel mutant. This class includes genes encoding putative stress proteins and global transcriptional regulators that might be

  19. Construction and structural analysis of integrated cellular network of Corynebacterium glutamicum%谷氨酸棒状杆菌集成细胞网络的构建与结构分析

    Institute of Scientific and Technical Information of China (English)

    姜金国; 宋理富; 郑平; 贾士儒; 孙际宾

    2012-01-01

    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 aad 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 hindered the systematic study of Corynebacterium glutamicum and large-scale rational design and optimization for strains. Here, by gathering relevant information from a number of public databases, we successfully constructed an integrated cellular network, which was composed of 1 384 reactions, 1 276 metabolites, 88 transcriptional factors and 999 pairs of transcriptional regulatory relationships. The transcriptional regulatory sub-network could be arranged into five layers and the metabolic sub-network presented a clear bow-tie structure. We proposed a new method to extract complex metabolic and regulatory sub-network for product-orientated study taking lysine biosynthesis as an example. The metabolic and regulatory sub-network extracted by our method was more close to the real functional network than the simplex biochemical pathways. The results would be greatly helpful for understanding the high-yielding biomechanism for amino acids and the re-design of the industrial strains.%谷氨酸棒状杆菌是一种重要的传统工业微生物,其基因组学和分子遗传操作工具的快速发展使得谷氨酸棒状杆菌具备了作为新型细胞工厂的潜力.但是,相对于大肠杆菌等模式生物,对于棒杆菌的代谢调控研究较少,特别是目前还缺乏谷氨酸棒状杆菌集成细胞网络的研究,这一现状阻碍了谷氨酸棒状杆菌的系统生物学研究和大规模菌种理性设计优化.文中综合应用公共数据库、文献数据库资源,

  20. Estudo da difteria na cidade do Recife. I. Nota sôbre levantamento de portadores de Corynebacterium diphtheriae no bairro dos Coelhos Survey on diphtheriae carriers in "Bairro dos Coelhos" Recife, Brazil

    Directory of Open Access Journals (Sweden)

    Dalva A. Mello

    1969-06-01

    Full Text Available De uma amostra probabilística do bairro dos Coelhos da cidade do Recife, 410 indivíduos foram examinados para verificação de portadores de difteria. Sòmente duas amostras de C. diphtheriae foram isoladas de duas crianças de 8 a 9 anos, as quais não apresentaram sintomatologia compatível com o quadro diftérico.From a limited population living around the University Hospital in Recife, Brazil a randomic sample was examined in order to identify diphtheria carriers. Swabs were made from 410 persons in a house-to-house survey. Two strains of Corynebacterium diphtheriae were isolated from healthy 8 and 9-year old children.

  1. l-Lysine production independent of the oxidative pentose phosphate pathway by Corynebacterium glutamicum with the Streptococcus mutans gapN gene.

    Science.gov (United States)

    Takeno, Seiki; Hori, Kazumasa; Ohtani, Sachiko; Mimura, Akinori; Mitsuhashi, Satoshi; Ikeda, Masato

    2016-09-01

    We have recently developed a Corynebacterium glutamicum strain that generates NADPH via the glycolytic pathway by replacing endogenous NAD-dependent glyceraldehyde 3-phosphate dehydrogenase (GapA) with a nonphosphorylating NADP-dependent glyceraldehyde 3-phosphate dehydrogenase (GapN) from Streptococcus mutans. Strain RE2, a suppressor mutant spontaneously isolated for its improved growth on glucose from the engineered strain, was proven to be a high-potential host for l-lysine production (Takeno et al., 2010). In this study, the suppressor mutation was identified to be a point mutation in rho encoding the transcription termination factor Rho. Strain RE2 still showed retarded growth despite the mutation rho696. Our strategy for reconciling improved growth with a high level of l-lysine production was to use GapA together with GapN only in the early growth phase, and subsequently shift this combination-type glycolysis to one that depends only on GapN in the rest of the growth phase. To achieve this, we expressed gapA under the myo-inositol-inducible promoter of iolT1 encoding a myo-inositol transporter in strain RE2. The resulting strain RE2A(iol) was engineered into an l-lysine producer by introduction of a plasmid carrying the desensitized lysC, followed by examination for culture conditions with myo-inositol supplementation. We found that as a higher concentration of myo-inositol was added to the seed culture, the following fermentation period became shorter while maintaining a high level of l-lysine production. This finally reached a fermentation period comparable to that of the control GapA strain, and yielded a 1.5-fold higher production rate compared with strain RE2. The transcript level of gapA, as well as the GapA activity, in the early growth phase increased in proportion to the myo-inositol concentration and then fell to low levels in the subsequent growth phase, indicating that improved growth was a result of increased GapA activity, especially in the

  2. l-Lysine production independent of the oxidative pentose phosphate pathway by Corynebacterium glutamicum with the Streptococcus mutans gapN gene.

    Science.gov (United States)

    Takeno, Seiki; Hori, Kazumasa; Ohtani, Sachiko; Mimura, Akinori; Mitsuhashi, Satoshi; Ikeda, Masato

    2016-09-01

    We have recently developed a Corynebacterium glutamicum strain that generates NADPH via the glycolytic pathway by replacing endogenous NAD-dependent glyceraldehyde 3-phosphate dehydrogenase (GapA) with a nonphosphorylating NADP-dependent glyceraldehyde 3-phosphate dehydrogenase (GapN) from Streptococcus mutans. Strain RE2, a suppressor mutant spontaneously isolated for its improved growth on glucose from the engineered strain, was proven to be a high-potential host for l-lysine production (Takeno et al., 2010). In this study, the suppressor mutation was identified to be a point mutation in rho encoding the transcription termination factor Rho. Strain RE2 still showed retarded growth despite the mutation rho696. Our strategy for reconciling improved growth with a high level of l-lysine production was to use GapA together with GapN only in the early growth phase, and subsequently shift this combination-type glycolysis to one that depends only on GapN in the rest of the growth phase. To achieve this, we expressed gapA under the myo-inositol-inducible promoter of iolT1 encoding a myo-inositol transporter in strain RE2. The resulting strain RE2A(iol) was engineered into an l-lysine producer by introduction of a plasmid carrying the desensitized lysC, followed by examination for culture conditions with myo-inositol supplementation. We found that as a higher concentration of myo-inositol was added to the seed culture, the following fermentation period became shorter while maintaining a high level of l-lysine production. This finally reached a fermentation period comparable to that of the control GapA strain, and yielded a 1.5-fold higher production rate compared with strain RE2. The transcript level of gapA, as well as the GapA activity, in the early growth phase increased in proportion to the myo-inositol concentration and then fell to low levels in the subsequent growth phase, indicating that improved growth was a result of increased GapA activity, especially in the

  3. Skin Infections Due to Corynebacterium

    OpenAIRE

    Meltem Türkmen; Derya Aytimur

    2010-01-01

    Corynebacteria are Gram-positive, non-sporulated, non-capsulated, aerobic diphtheroid bacteria accounting for nearly 50%of the natural skin biocene. This bacterial family is responsible for various skin diseases such as cutaneous diphteria, cromhydrosis, bromhydrosis but the most common of them are pitted keratolysis, trichobacteriosis and erythrasma. A warm and moist environment and poor hygiene are the predisposition factors for these three diseases. Although this skin diseases are seen mor...

  4. 小麦淀粉糖发酵过程谷氨酸菌理化性质研究%Research on Physical and Chemical Properties of Corynebacterium glutamicum in Fermentation Process of the Glucose of Wheat Starch

    Institute of Scientific and Technical Information of China (English)

    黄继红; 周靖波; 欧歌

    2013-01-01

    [目的]为了考察小麦淀粉糖发酵过程中谷氨酸菌种的理化特性.[方法]以不同发酵时间内代谢谷氨酸的纯度以及菌体的光密度和细胞活性能为检测指标、并对定性和定量指标进行分析回归比较.[结果]华勃氏微量呼吸仪与双功能分析仪在试验范围内,检测误差为d=3.316 6%,其线性关系为s1=(1-d)s2,在小麦淀粉糖发酵过程中,细胞活性能平均值6.24μA,最高值为6.61 μA.[结论]细胞活性能比菌体光密度更真实反应菌体在小麦淀粉糖发酵过程的理化性质,细胞膜磷脂与细胞活性能有相关性.%This study aimed to investigate the physical and chemical characteristic of Corynebacterium glutamicum in fermentation process of the glucose of wheat starch.[Method] The purity of glutamic acid in fermentation period,optical density and cell viability of bacteria were detected as indicators for regression comparison and analysis.[Result] The relative error d=3.316 6% within the experimental range of Warburg trace breathing apparatus and double function analyzer.The linear relationship was s1=(1-d)s2.During the fermentation process of the glucose of wheat starch,the average cell activity was 6.24 μA and the maximum cell activity was 6.61 μA.[Conclusion] Compared with optical density,cell viability can more accurately reflect the physical and chemical properties of Corynebacterium glutamicum in fermentation process of the glucose of wheat starch.There was certain correlation between cell membrane phospholipids and cell viability.

  5. Genetic relationships of Corynebacterium diphtheriae strains isolated from a diphtheria case and carriers by restriction fragment length polymorphism of rRNA genes Relação genética de cepas de Corynebacterium diphtheriae isoladas de caso e seus contatos por RLFP de rRNA gene

    Directory of Open Access Journals (Sweden)

    Claudio Tavares Sacchi

    1995-08-01

    Full Text Available In the present study we report the results of an analysis, based on ribotyping of Corynebacterium diphtheriae intermedius strains isolated from a 9 years old child with clinical diphtheria and his 5 contacts. Quantitative analysis of RFLPs of rRNA was used to determine relatedness of these 7 C.diphtheriae strains providing support data in the diphtheria epidemiology. We have also tested those strains for toxigenicity in vitro by using the Elek's gel diffusion method and in vivo by using cell culture method on cultured monkey kidney cell (VERO cells. The hybridization results revealed that the 5 C.diphtheriae strains isolated from contacts and one isolated from the clinical case (nose case strain had identical RFLP patterns with all 4 restriction endonucleases used, ribotype B. The genetic distance from this ribotype and ribotype A (throat case strain, that we initially assumed to be responsible for the illness of the patient, was of 0.450 showing poor genetic correlation among these two ribotypes. We found no significant differences concerned to the toxin production by using the cell culture method. In conclusion, the use of RFLPs of rRNA gene was successful in detecting minor differences in closely related toxigenic C.diphtheriae intermedius strains and providing information about genetic relationships among them.No presente estudo, nós reportamos os resultados de uma análise, baseada na ribotipagem de cepas de C. diphtheriae intermedius isoladas de uma criança de 9 anos com difteria e seus 5 contatos. Análise quantitativa por RFLP de rRNA foi usada para determinar a relação destas 7 cepas de C. diphtheriae fornecendo dados de interesse epidemiológico. Nós também testamos estas cepas para toxicidade in vitro usando método de difusão de Elek e in vivo usando método de cultura celular com células VERO. Os resultados de hibridização revelaram que as 5 cepas de C. diphtheriae isoladas dos contatos e uma isolada do caso (cepa isolada

  6. Corynebacterium glutamicum MTCC 2745 immobilized on granular activated carbon/MnFe2O4 composite: A novel biosorbent for removal of As(III) and As(V) ions.

    Science.gov (United States)

    Podder, M S; Majumder, C B

    2016-11-01

    The optimization of biosorption/bioaccumulation process of both As(III) and As(V) has been investigated by using the biosorbent; biofilm of Corynebacterium glutamicum MTCC 2745 supported on granular activated carbon/MnFe2O4 composite (MGAC). The presence of functional groups on the cell wall surface of the biomass that may interact with the metal ions was proved by FT-IR. To determine the most appropriate correlation for the equilibrium curves employing the procedure of the non-linear regression for curve fitting analysis, isotherm studies were performed for As(III) and As(V) using 30 isotherm models. The pattern of biosorption/bioaccumulation fitted well with Vieth-Sladek isotherm model for As(III) and Brouers-Sotolongo and Fritz-Schlunder-V isotherm models for As(V). The maximum biosorption/bioaccumulation capacity estimated using Langmuir model were 2584.668mg/g for As(III) and 2651.675mg/g for As(V) at 30°C temperature and 220min contact time. The results showed that As(III) and As(V) removal was strongly pH-dependent with an optimum pH value of 7.0. D-R isotherm studies specified that ion exchange might play a prominent role. PMID:27289352

  7. Role of Key Residues at the Flavin Mononucleotide (FMN:Adenylyltransferase Catalytic Site of the Bifunctional Riboflavin Kinase/Flavin Adenine Dinucleotide (FAD  Synthetase from Corynebacterium ammoniagenes

    Directory of Open Access Journals (Sweden)

    Susana Frago

    2012-11-01

    Full Text Available In mammals and in yeast the conversion of Riboflavin (RF into flavin mononucleotide (FMN and flavin adenine dinucleotide (FAD is catalysed by the sequential action of two enzymes: an ATP:riboflavin kinase (RFK and an ATP:FMN adenylyltransferase (FMNAT. However, most prokaryotes depend on a single bifunctional enzyme, FAD synthetase (FADS, which folds into two modules: the C-terminal associated with RFK activity and the N-terminal associated with FMNAT activity. Sequence and structural analysis suggest that the 28-HxGH-31, 123-Gx(D/N-125 and 161-xxSSTxxR-168 motifs from FADS must be involved in ATP stabilisation for the adenylylation of FMN, as well as in FAD stabilisation for FAD phyrophosphorolysis. Mutants were produced at these motifs in the Corynebacterium ammoniagenes FADS (CaFADS. Their effects on the kinetic parameters of CaFADS activities (RFK, FMNAT and FAD pyrophosphorilase, and on substrates and product binding properties indicate that H28, H31, N125 and S164 contribute to the geometry of the catalytically competent complexes at the FMNAT-module of CaFADS.

  8. Metabolic Engineering in Corynebacterium glutamicum to Increase L-Valine Production%谷氨酸棒杆菌生产缬氨酸的代谢工程研究进展

    Institute of Scientific and Technical Information of China (English)

    王小元

    2012-01-01

    L-缬氨酸是人体必需的三种支链氨基酸之一,在生命代谢过程中起着非常重要的作用,因此被广泛应用于食品、医药及饲料等行业.目前,L-缬氨酸主要采用微生物发酵法生产,而谷氨酸棒杆菌是最常用的生产菌种.作者分析了谷氨酸棒杆菌中L-缬氨酸的生物合成途径和代谢调控,综述了对其进行代谢工程改造来提高L-缬氨酸产量的最新研究进展.%L-valine, one of the three branched-chain amino acids, is essential for human, and plays an important role in the metabolism of life, therefore, it is widely used in products of food, medicine and feed. L-valine is mainly produced by bacterial fermentation, and the most used bacterium is Corynebacterium glutamicum. In this article, the pathway and regulation of L-valine biosynthesis in C. glutamicum are analyzed and researches on metabolic engineering to increase L-valine production in C. glutamicum are reviewed.

  9. Factors enhancing L-valine production by the growth-limited L-isoleucine auxotrophic strain Corynebacterium glutamicum DeltailvA DeltapanB ilvNM13 (pECKAilvBNC).

    Science.gov (United States)

    Denina, Ilze; Paegle, Longina; Prouza, Marek; Holátko, Jiri; Pátek, Miroslav; Nesvera, Jan; Ruklisha, Maija

    2010-07-01

    Cell growth limitation is known to be an important condition that enhances L: -valine synthesis in Corynebacterium glutamicum recombinant strains with L: -isoleucine auxotrophy. To identify whether it is the limited availability of L: -isoleucine itself or the L: -isoleucine limitation-induced rel-dependent ppGpp-mediated stringent response that is essential for the enhancement of L: -valine synthesis in growth-limited C. glutamicum cells, we deleted the rel gene, thereby constructing a relaxed (rel (-) ) C. glutamicum DeltailvA DeltapanB Deltarel ilvNM13 (pECKAilvBNC) strain. Variations in enzyme activity and L: -valine synthesis in rel (+) and rel (-) strains under conditions of L: -isoleucine excess and limitation were investigated. A sharp increase in acetohydroxy acid synthase (AHAS) activity, a slight increase in acetohydroxyacid isomeroreductase (AHAIR) activity, and a dramatic increase in L: -valine synthesis were observed in both rel (+) and rel (-) cells exposed to L: -isoleucine limitation. Although the positive effect of induction of the stringent response on AHAS and AHAIR upregulation in cells was not confirmed, we found the stringent response to be beneficial for maintaining increased AHAS, dihydroxyacid dehydratase, and transaminase B activity and L: -valine synthesis in cells during the stationary growth phase.

  10. 短小棒状杆菌CpG-DNA提取方法的研究%Studied on the method of extracting CpG-DNA from Corynebacterium parvum

    Institute of Scientific and Technical Information of China (English)

    孙彬裕; 高尚先

    2009-01-01

    目的:建立一种高效纯化CP-CpG-DNA的方法,为研究DNA免疫增强佐剂功能奠定基础.方法:应用经超声破碎、蛋白酶K消化、CTAB沉淀及酚、氯仿抽提等步骤从短棒杆菌(Corynebacterium parvum,CP)中提取基因组DNA(CP-CpG-DNA).结果:琼脂糖凝胶电泳检测,CP-CpG-DNA主要集中在2.3 Mb;CP-CpG-DNA经限制性内切酶HpaⅡ酶切后,琼脂糖凝胶电泳检测,得到小于1000bp的片断.结论:建立的高效纯化CP-CpG-DNA方法.获得的CP-CpG-DNA纯度较好,而且CP-CpG-DNA中存在大量非甲基化CpG位点,可用于DNA免疫增强佐剂的应用.

  11. Corynebacterium parvum- and Mycobacterium bovis bacillus Calmette-Guerin-induced granuloma formation is inhibited in TNF receptor I (TNF-RI) knockout mice and by treatment with soluble TNF-RI.

    Science.gov (United States)

    Senaldi, G; Yin, S; Shaklee, C L; Piguet, P F; Mak, T W; Ulich, T R

    1996-12-01

    The aim of this study was to examine the role of TNF receptor I (TNF-RI) in the pathogenesis of heat-killed Corynebacterium parvum- and live bacillus Calmette-Guerin (BCG)-induced granulomas. Granuloma formation was analyzed in TNF-RI knockout mice and after treatment with soluble TNF-RI (sTNF-RI). TNF-RI knockout mice injected with C. parvum or BCG developed fewer and smaller granulomas than wild-type control mice. Mice treated with sTNF-RI from days 7 to 13 after injection of C. parvum or BCG developed fewer and smaller granulomas than saline-treated control mice. Established granulomas regressed in rats treated with sTNF-RI from days 10 to 13 after injection of C. parvum. In conclusion, TNF signaling via TNF-RI contributes to the pathogenesis of C. parvum- and BCG-induced granulomas. sTNF-RI inhibits the development of granulomas and can cause the regression of established granulomas. PMID:8943410

  12. Corynebacterium glutamicum MTCC 2745 immobilized on granular activated carbon/MnFe2O4 composite: A novel biosorbent for removal of As(III) and As(V) ions

    Science.gov (United States)

    Podder, M. S.; Majumder, C. B.

    2016-11-01

    The optimization of biosorption/bioaccumulation process of both As(III) and As(V) has been investigated by using the biosorbent; biofilm of Corynebacterium glutamicum MTCC 2745 supported on granular activated carbon/MnFe2O4 composite (MGAC). The presence of functional groups on the cell wall surface of the biomass that may interact with the metal ions was proved by FT-IR. To determine the most appropriate correlation for the equilibrium curves employing the procedure of the non-linear regression for curve fitting analysis, isotherm studies were performed for As(III) and As(V) using 30 isotherm models. The pattern of biosorption/bioaccumulation fitted well with Vieth-Sladek isotherm model for As(III) and Brouers-Sotolongo and Fritz-Schlunder-V isotherm models for As(V). The maximum biosorption/bioaccumulation capacity estimated using Langmuir model were 2584.668 mg/g for As(III) and 2651.675 mg/g for As(V) at 30 °C temperature and 220 min contact time. The results showed that As(III) and As(V) removal was strongly pH-dependent with an optimum pH value of 7.0. D-R isotherm studies specified that ion exchange might play a prominent role.

  13. Corynebacterium glutamicum MTCC 2745 immobilized on granular activated carbon/MnFe2O4 composite: A novel biosorbent for removal of As(III) and As(V) ions.

    Science.gov (United States)

    Podder, M S; Majumder, C B

    2016-11-01

    The optimization of biosorption/bioaccumulation process of both As(III) and As(V) has been investigated by using the biosorbent; biofilm of Corynebacterium glutamicum MTCC 2745 supported on granular activated carbon/MnFe2O4 composite (MGAC). The presence of functional groups on the cell wall surface of the biomass that may interact with the metal ions was proved by FT-IR. To determine the most appropriate correlation for the equilibrium curves employing the procedure of the non-linear regression for curve fitting analysis, isotherm studies were performed for As(III) and As(V) using 30 isotherm models. The pattern of biosorption/bioaccumulation fitted well with Vieth-Sladek isotherm model for As(III) and Brouers-Sotolongo and Fritz-Schlunder-V isotherm models for As(V). The maximum biosorption/bioaccumulation capacity estimated using Langmuir model were 2584.668mg/g for As(III) and 2651.675mg/g for As(V) at 30°C temperature and 220min contact time. The results showed that As(III) and As(V) removal was strongly pH-dependent with an optimum pH value of 7.0. D-R isotherm studies specified that ion exchange might play a prominent role.

  14. Scientific Opinion on the safety and efficacy of L-valine (ValAMINO® produced by Corynebacterium glutamicum (DSM 25202 for all animal species, based on a dossier submitted by Evonik Industries AG

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2014-07-01

    Full Text Available The product L-valine, feed grade (ValAMINO®, is a feed additive produced by fermentation with a genetically modified strain of Corynebacterium glutamicum (DSM 25202. Neither the production strain nor its recombinant DNA was detected in the final product. Therefore, the final product does not give rise to any safety concerns with regard to the genetic modification. The additive L-valine, feed grade, produced by C. glutamicum (DSM 25202 is safe for all target animals when supplemented in appropriate amounts to feed to compensate for valine deficiency in feedingstuffs. The composition of tissues and products of animal origin will not be changed by the use of L-valine, feed grade, in animal nutrition. Considering the high purity of the product under assessment, no risks to the consumer are expected from its use as a feed additive. In the absence of specific data on user safety for the product under application, it would be prudent to consider it as an irritant to the skin, eyes and mucous membranes and as a potential dermal sensitiser. In the absence of any data on inhalation toxicity, the product is also regarded as potentially harmful by inhalation. The amino acid L-valine is a natural component of proteins of living organisms. Therefore, the use of the highly purified product L-valine, feed grade, in animal nutrition does not pose a risk to the environment. The product L-valine, feed grade, is an efficacious source of supplemented amino acid for all species, but it has a low efficiency in ruminants unless it is used in a protected form.

  15. The crystal structures of apo and cAMP-bound GlxR from Corynebacterium glutamicum reveal structural and dynamic changes upon cAMP binding in CRP/FNR family transcription factors.

    Directory of Open Access Journals (Sweden)

    Philip D Townsend

    Full Text Available The cyclic AMP-dependent transcriptional regulator GlxR from Corynebacterium glutamicum is a member of the super-family of CRP/FNR (cyclic AMP receptor protein/fumarate and nitrate reduction regulator transcriptional regulators that play central roles in bacterial metabolic regulatory networks. In C. glutamicum, which is widely used for the industrial production of amino acids and serves as a non-pathogenic model organism for members of the Corynebacteriales including Mycobacterium tuberculosis, the GlxR homodimer controls the transcription of a large number of genes involved in carbon metabolism. GlxR therefore represents a key target for understanding the regulation and coordination of C. glutamicum metabolism. Here we investigate cylic AMP and DNA binding of GlxR from C. glutamicum and describe the crystal structures of apo GlxR determined at a resolution of 2.5 Å, and two crystal forms of holo GlxR at resolutions of 2.38 and 1.82 Å, respectively. The detailed structural analysis and comparison of GlxR with CRP reveals that the protein undergoes a distinctive conformational change upon cyclic AMP binding leading to a dimer structure more compatible to DNA-binding. As the two binding sites in the GlxR homodimer are structurally identical dynamic changes upon binding of the first ligand are responsible for the allosteric behavior. The results presented here show how dynamic and structural changes in GlxR lead to optimization of orientation and distance of its two DNA-binding helices for optimal DNA recognition.

  16. Effect of enzyme-digested Corynebacterium parvum product on immune function of tumor-bearing mice%酶法处理短棒状杆菌对荷瘤鼠脾细胞免疫功能的影响

    Institute of Scientific and Technical Information of China (English)

    曹芳; 杜瑞林; 郭立君

    2012-01-01

    Objective To investigate the effect of enzyme-digested Corynebacterium parvum product (ECPP) on immune function of lymphocytes of tumor-bearing mice. Methods NIH mice were randomly divided into saline control (NS), tumor-bearing control (CK), positive control (Corynebacterium parvum product, CPP) as well as low (ECPP Ⅰ , 500 μg/ml) and high (ECPP Ⅱ, 1 000 μg/ml) dose ECPP groups. The mice in NS group were injected i.p. with 0. 2 ml of 0. 9% NaCl, while those in other four groups with 0. 2 ml of Ehrlich ascites carcinoma (EAC) cell suspension (5. 0×106 cells/ml). The mice in CPP, ECPP Ⅰ and ECPP Ⅱ groups were given the corresponding drugs (each at a volume of 0. 25 ml) starting from the 2nd day after inoculation of EAC cells ev-ery other day for 5 times, then killed on the 2nd day after withdrawal, and determined for malignant peritoneal effusion. The spleen lymphocyte transformation level was determined by 3H-TdR method, while the T lymphocyte subgroup and activity of national killer (NK) cells by flow cytometry, and the transcription levels of IFNγ and TNF-α mRNAs by RT-PCR. Results Compared with those in CK group, the malignant peritoneal effusion levels in CPP, ECPP Ⅰ and ECPP Ⅱ groups decreased significantly (P 0. 05), and was significantly low-er in ECPPⅡ than in ECPP I group (P 0. 05). Conclusion ECPP enhanced the immune function of splenic lymphocytes of tumor-bearing mice.%目的 探讨酶法处理短棒状杆菌(Enzyme-digested Corynebacterium parvum product,ECPP)对荷瘤鼠脾淋巴细胞免疫功能的影响.方法 将NIH小鼠随机分为5组:生理盐水对照组(NS)、荷瘤对照组(CK)、阳性对照组(CPP)、低剂量ECPP组(ECPPⅠ,500 μg/ml ECPP)和高剂量ECPP组(ECPPⅡ,1 000 μg/ml ECPP).除NS组经腹腔注射0.2 ml 0.9% NaCl外,其他4组小鼠均经腹腔注射0.2ml艾氏腹水瘤(Ehrlich ascites carcinoma,EAC)细胞悬液(5.0×106个/ml),接种次日,每组小鼠腹腔注射相应药物(注射体积均为0.25 ml

  17. INFLUENCE OF THE NON-CELL NANO SCALE PRODUCT FROM CORYNEBACTERIUM PARVUM(NCPP) ON NO PRODUCED BY THE PHAGOCYTOSIS OF MACROPHAGES%无细胞短棒状杆菌纳米级制剂对小鼠腹腔巨噬细胞产生NO的影响

    Institute of Scientific and Technical Information of China (English)

    李玮; 张利宁; 宋静; 高尚先; 王群; 张萍

    2004-01-01

    目的:研究无细胞棒状杆菌纳米制剂(Non-cell Nano Scale Product from Corynebacterium Parvum,NCPP)对小鼠腹腔巨噬细胞(PMψ)产生一氧化氮(NO)的影响,探讨其抗肿瘤作用机制,为临床应用提供依据.方法:采用小鼠腹腔注射NCPP与短棒状杆菌制剂(Corynebacterium Parvum Product,CPP),10 d后检测NO水平发现NCPP具有与CPP相同的作用.结果:NCPP组和CPP组与对照组比较,检测结果显示,NCPP和CPP均可明显提高小鼠腹腔Mψ产生NO的水平,两组与对照组相比,有极显著性差异(P<0.01),但NCPP和CPP两组之间无显著性差异(P>0.05).结论:NCPP与CPP具有相同作用,可激活巨噬细胞,分泌产生高水平NO以达到杀伤肿瘤细胞的目的.

  18. 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

    Directory of Open Access Journals (Sweden)

    Busche Tobias

    2012-09-01

    Full Text Available Abstract Background 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 effects of RshA on the SigH regulon, in order to refine the model describing the role of SigH and RshA during stress response. Results Transcription analyses revealed that the sigH gene and rshA gene are cotranscribed from four sigH housekeeping promoters in C. glutamicum. In addition, a SigH-controlled rshA promoter was found to only drive the transcription of the rshA gene. To test the role of the putative anti-sigma factor gene rshA under normal growth conditions, a C. glutamicum rshA deletion strain was constructed and used for genome-wide transcription profiling with DNA microarrays. In total, 83 genes organized in 61 putative transcriptional units, including those previously detected using sigH mutant strains, exhibited increased transcript levels in the rshA deletion mutant compared to its parental strain. The genes encoding proteins related to disulphide stress response, heat stress proteins, components of the SOS-response to DNA damage and proteasome components were the most markedly upregulated gene groups. Altogether six SigH-dependent promoters upstream of the identified genes were determined by primer extension and a refined consensus promoter consisting of 45 original promoter sequences was constructed. Conclusions The rshA gene codes for an anti-sigma factor controlling the function of the stress-responsive sigma factor SigH in C. glutamicum. Transcription of rshA from a SigH-dependent promoter may serve to quickly

  19. 有机氮源对谷氨酸棒杆菌发酵L-缬氨酸的影响%The Effects of Organic Nitrogen Sources on the Fermentation of L-valine by Corynebacterium glutamicum

    Institute of Scientific and Technical Information of China (English)

    徐庆阳; 孙家凯; 吴晓娇; 王晶; 谢希贤; 陈宁

    2012-01-01

    以L-缬氨酸生产菌谷氨酸棒杆菌XV0505为供试菌株,研究有机氮源对L-缬氨酸发酵的影响,确定了玉米浆代替豆饼水解液作为有机氮源的发酵工艺,降低了发酵成本;考察不同玉米浆浓度对谷氨酸棒杆菌XV0505发酵生产工一缬氨酸过程中生物量、耗糖速率、L-缬氨酸产量、副产物积累及氨消耗等方面影响,确定了玉米浆的适宜添加浓度;考察了玉米浆与生物素不同配比对L-缬氨酸分批发酵过程的影响,确定了最适生物素添加浓度。与原工艺相比,新工艺的菌体生物量及产酸提高了13.2%和18.5%。%The effects of organic nitrogen sources on the fermentation of L-valine were studied with L-valine-pro-ducing strain Corynebacterium glutamicum XV0505. Corn steep liquor was selected as the appropriate nitrogen source instead of soybean hydrolysates to reduce the cost of fermentation. The effects of initial concentration of corn steep liq-uor in fermentation process on biomass, glucose consumption rate, yield of L-valine, accumulation of byproduct and ammonia consumption rate were studied by carrying out fed-batch fermentation. The optimal initial concentration of corn steep liquor was also determined. The effects of the combined agents, biotin and corn steep liquor, on overpro-duction of L-valine were also studied, and the concentration of biotin was optimized. Compared with those of the origi-nal process, the biomass and L-valine production from the improved process were increased by 13.2% and 18.5 % respectively.

  20. 细菌16 S核糖体RNA基因检测杰氏棒状杆菌一株%Identification of corynebacterium jeikeium by detection of bacterial 16s ribosomal RNA gene

    Institute of Scientific and Technical Information of China (English)

    崔颖鹏; 黄朱亮

    2015-01-01

    Objective To detect clinical strain by amplifying 16S ribosomal RNA( 16S rRNA) gene with polymerase chain reaction ( PCR) method. Methods 16S rRNA gene was amplifyied by universal primers P1,P2 with polymerase chain reaction,and the polymerase chain reaction products were sequenced to achieve the DNA sequences. Results PCR products were about 1 465bp, and the PCR sequences were blasted in NCBI web to achieve the proper strain name,with the use of this method,the unkown gram⁃positive bacilli strain was identified as corynebacterium jeikeium strain, ATCC25923 was identified as a strain of staphylococcus aureus and negative control was also specific.Conclusion 16S rRNA gene detection can be used as strain identification,and especially for some less common clinical strains.%目的:探讨通过16S核糖体RNA(16S rRNA )基因测序的方法用于临床菌株鉴定。方法以细菌16S rRNA基因为靶序列,采用细菌的通用引物P1、P2进行PCR扩增,同时送到测序公司测序从而达到鉴定。结果待测菌株及ATCC25923阳性对照通过PCR得到的扩增片段为1465bp左右,待测菌株经测序比对鉴定为一株杰氏棒状杆菌,阳性质控ATCC25923鉴定为一株金黄的葡萄球菌,阴性对照未出现任何结果。结论通过PCR检测细菌16srRNA基因可以应用于临床菌株的鉴定,并且适用一些难以鉴定的细菌。

  1. Formation of volutin granules in Corynebacterium glutamicum.

    Science.gov (United States)

    Pallerla, Srinivas Reddy; Knebel, Sandra; Polen, Tino; Klauth, Peter; Hollender, Juliane; Wendisch, Volker F; Schoberth, Siegfried M

    2005-02-01

    Volutin granules are intracellular storages of complexed inorganic polyphosphate (poly P). Histochemical staining procedures differentiate between pathogenic corynebacteria such as Corynebacterum diphtheriae (containing volutin) and non-pathogenic species, such as C. glutamicum. Here we report that strains ATCC13032 and MH20-22B of the non-pathogenic C. glutamicum also formed subcellular entities (18-37% of the total cell volume) that had the typical characteristics of volutin granules: (i) volutin staining, (ii) green UV fluorescence when stained with 4',6-diamidino-2-phenylindole, (iii) electron-dense and rich in phosphorus when determined with transmission electron microscopy and X-ray microanalysis, and (iv) 31P NMR poly P resonances of isolated granules dissolved in EDTA. MgCl2 addition to the growth medium stimulated granule formation but did not effect expression of genes involved in poly P metabolism. Granular volutin fractions from lysed cells contained polyphosphate glucokinase as detected by SDS-PAGE/MALDI-TOF, indicating that this poly P metabolizing enzyme is present also in intact poly P granules. The results suggest that formation of volutin is a more widespread phenomenon than generally accepted. PMID:15668011

  2. Optimization of lysine metabolism in Corynebacterium glutamicum

    DEFF Research Database (Denmark)

    Rytter, Jakob Vang

    , and increased NADPH availability is therefore a potential way to enhance lysine production. The generation of NADPH is mainly located in the pentose phosphate pathway (PPP). Using the genome scale model the phosphoglucoisomerase enzyme (PGI) has been identified as a possible bottleneck in the metabolism, which...

  3. 原位氮饥饿发酵工艺中梯度补氮对谷氨酰胺合成酶的调控%REGULATION OF GLUTAMINE SYNTHETASE IN GLUTAMINE PRODUCTION BY FERMENTATION OF Corynebacterium glutamicum WITH in-situ NITROGEN STARVATION AND GRADIENT FED NITROGEN

    Institute of Scientific and Technical Information of China (English)

    李春; 刘雨磊; 陈奎发; 杨艳; 曹竹安

    2004-01-01

    The effects of uniform and gradient fed nitrogen on glutamine synthetase (GS), glutamate dehydrogenase(GDH) and glutamate synthase ((K)GAT)were investigated in glutamine production by fermentation of Corynebacterium glutamicum NS611 after 3 h of in-situ nitrogen starvation. It was shown that the strain in the later growth phase entered naturally into in-situ nitrogen starvation by controlling the initial concentration of urea and the biomass was slightly decreased. The pH value reached 6.5 again in the culture system, which confirmed the beginning of nitrogen starvation in the culture system. After 3 h nitrogen starvation the activity of GS was increased over two folds and the time of high activity of GS persisted three folds longer in the gradient fed nitrogen system than that in the normal fed batch. The higher activity of GDH was also maintained. The glutamine production increased by 72 % than the original technology of nitrogen starvation and the time of fermentation was shortened by above 12 h.

  4. 陕南白山羊伪结核棒状杆菌分离鉴定及灭活铝胶疫苗研制%Isolation and Identification of Corynebacterium pseudotuberculosis from White Goat in South Shaanxi and Development of an Inactivated Vaccine with Aluminum Hydroxide Adjuvant

    Institute of Scientific and Technical Information of China (English)

    蔡俊波; 王爱玲; 李海敏; 李春燕; 焦阳阳; 张彦明; 郭抗抗

    2016-01-01

    Caseous lymphadenitis also known as ovine pseudotuberculosis,is a chronic zoonotic infectious disease caused by Corynebacterium pseudotuberculosis .This disease is harmful to goat especially,and there is not yet an effective treatment to prevent this disease.White goat is a main breed in Shiquan county of south Shaanxi,Corynebacterium pseudotuberculosis is an important pathogen in goat population and causes large economic losses in recent years.In order to screen drugs that are sensitive to local Corynebacterium pseudotuberculosis and develop an inactivated vaccine,the fresh pus were collected from the suspected pseudotuberculosis goats,the bacteria were isolated and identified from those samples.And then,drug sen-sitivity test was carried out with 1 isolate,and then an inactivated vaccine with aluminum hydroxide adju-vant was developed based this isolate.The results showed that the isolated bacteria formed a medium sized white,dry,flat and opaque,neat edge colony at solid agar,showing Gram stain positive.The isolate was confirmed as Corynebacterium pseudotuberculosis by analysis of 1 6 S rRNA sequences.Susceptibility test results showed this isolate was highly sensitive to ciprofloxacin,ceftriaxone,erythromycin,kanamycin,ce-fotaxime,moderately sensitive to tobramycin,doxycycline,gentamicin,chloramphenicol;tetracycline,rif-ampicin,streptomycin.Subsequently,an activated vaccine was developed with aluminum hydroxide adjuvant based on this isolate,no live bacterium was detected in this vaccine.The inactivated vaccine was injected to experimental goats in those herds that the samples were collected,and no abnormal reactions were ob-served in the goats.Clinical application also was carried out in goat herd and the immune effects will be e-valuated later.This study provided an experimental basis for effective drugs and a choice of inactivated vac-cine for preventing and controlling caseous lymphadenitis.%羊干酪性淋巴结炎又称羊伪结核病,是由伪结核棒

  5. MICROBIAL LANDSCAPE OF MICROFLORA OF A PHARYNX AT PATIENTS WITH TONSILLIT’S PATHOLOGY

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    O. Yu. Borisova

    2015-01-01

    Full Text Available The microbial landscape of microflora of a pharynx at patients with tonsillit’s pathology were studied. Materials and methods. 79 patients from GBUZ “Research Institute of Clinical Otorhinolaryngology” (78.5% patients with various forms of chronic tonsillitis and 21.5% of patients without chronic tonsillitis (control group. Among patients of the main group with chronic tonsillitis, at 60 (96.8% patients there was a diagnosis chronic tonsillitis a toxical-allergic form of 1 degree (TAF I, at 1 (1.6% the patient — chronic tonsillitis a toxical-allergic form II of degree (TAF II and at 1 (1.6% the patient — chronic tonsillitis. Identification of microorganisms carried out on cultural-morphological and biochemical properties. Specific identification of the hardly cultivated microorganisms and Corynebacterium was carried out by MALDI-TOFF MS of BioMerieux VITEK MS MALDI-TOF (“bioMerieux”, France. Identification of the allocated Corynebacterium was carried out by amplification of a gene rpoB and the subsequent direct sequencing. Results. The majority (98.7% of the allocated microorganisms, treated 27 types and were Gram-positive. It is revealed 159 strains of 29 species of microorganisms, from them 41.4% of strains belonged to the Streptococcus, 19.7% — Staphylococcus, 36.9% — Corynebacterium. Among Streptococcus — 55.4% of S. viridans, 38.4% — S. pyogenes and 3.1% — of S. pneumoniae и S. oralis; Staphylococcus — 64.5% of S. aureus, 32.2% of S. epidermidis and 3.3% of S. hominis. 18 types of Corynebacterium — С. tuberculostearicum (17.2% strains, C. рseudodiphtheriticum (15.5% strains and C. aurimucosum (18.9% strains are revealed. At 44.3% of the surveyed patients the microflora is presented by a monoculture and at 55.7% associations are revealed. Conclusion. The microflora at patients with tonsillit’s pathology is characterized. At the expressed pathological process in a microbiota of a pharynx the monoculture while

  6. 谷氨酸棒杆菌L-色氨酸营养缺陷型突变及其高产菌株的选育%Study on breeding of Corynebacterium glutamicum L-tryptophan auxotrophic strains with high yield

    Institute of Scientific and Technical Information of China (English)

    张新武; 侯钢北; 杨晓明; 廉娜娜

    2015-01-01

    Objective To research the breeding of high L-tryptophan yield with multiple synthetic and metabolic pathway mutation strain by using Corynebacterium glutamicum HX-22 as the starting strain. Methods Diethyl sulfate (DES), UV, and cobalt-60 gamma-ray mutagenesis method were performed cross processing starting on C. glutamicum HX-22 and mutant strain. The auxotrophic phenotype and metabolic suicide substrate resistance screening methods were used, and targeted mutagenesis and breeding of L-tryptophan acid synthesis and catabolism mutation of lubricious ammonia acid producing strain. Results Through continuous type defect mutagenesis breeding for many times, the strains with auxotroph and resistance marker of L-tryptophan high yield bacteria HX22-118 (Phe--Tyr--5FTr-4FPr-SGr) was screened, then extend the 10 times in succeeding transfer culture, the L-trp fermentation yield reached to 27.1 g/L, which was increased by 81.8% comparing to the wild start strain. Conclusion The selected multiple mutated strain HX22-118 (Phe--Tyr--5FTr-4FPr-SGr) with high L-trp yielding strain has a good genetic stability, which can be applied in industrial application.%目的:以谷氨酸棒杆菌(Corynebacterium glutamicum)HX-22为出发菌株,研究了目标发酵产物L-色氨酸合成途径交叉反馈抑制途径代谢突变与色氨酸分解代谢类似物抗性的营养缺陷型突变诱导过程及 L-色氨酸高产复合突变菌株的筛选。方法采用硫酸二乙酯、紫外线、钴60γ-射线等诱变方法交叉处理起始与突变菌株,通过营养缺陷表型和自杀代谢底物抗性的筛选方法,定向突变和选育 L-色氨酸合成与分解代谢突变的色氨酸高产菌株。结果通过多次连续营养缺陷型诱变选育,筛选出一株具有分支酸-Trp/Phe/Tyr代谢途径L-Phe和 L-Tyr 合成缺陷型和 L-Trp 分解代谢自杀代谢底物抗性的 L-色氨酸高产菌 HX22-118(Phe--Tyr--5FTr-4FPr-SGr);通过连续传代10次

  7. 钝齿棒杆菌argR基因克隆、表达及其重组菌发酵产精氨酸的探讨%Study on Gene cloning and expression of argR and its recombinant strain producing arg through fermentation in Corynebacterium crenatum

    Institute of Scientific and Technical Information of China (English)

    石磊

    2013-01-01

    目的 探讨N-乙酰鸟氨酸的转氨酶于钝齿棒杆菌当中,其精氨酸合成环节所起到的作用,并验证其酶学性质,从而为优化提高培养基的成分,以及发酵过程的工艺条件,和精氨酸的产量提供理论及实验依据.方法 于精氨酸的高产菌株中,使其钝齿棒杆菌的染色体(SYPA5-5)扩增,从而获取ACOAT的编码基因-argD,其全长为1176bp,编码氨基酸为390个,均于C.crenatum SYPA与Escherichia coli BL21(DE3)上成功表达.选择Ni柱进行亲和层析并纯化后,能够得到重组的蛋白比酶活可达到108.2U/g,并研究其酶学性质.再构建重组的钝齿棒杆菌,以加强其精氨酸的合成途径中ACOAT的蛋白表达量,同时对重组的菌产精氨酸予以发酵分析.结果 重组的钝齿棒杆菌同出发菌株进行对比,其胞内的ACOAT的酶活得到增强;并且重组菌中CCD1其精氨酸的平均产量达到了39.7g/L,其产酸最终提高了14.7%.结论 通过研究,不仅对高产精氨酸其重组基因菌工程的构建提供以理论依据,而且还具有非常重要的指导意义.%Objective N - acety - L - omithine transaminase which reacted at the step of arg biosynthesis was not only discussed, but also its enzyme properties were proved. It even could provide theoretical and experimental basis for improving the culture, optimizing technological conditions and increasing output of Arg. Methods Through amplifying chromosome SYPA5 -5 of Corynebacterium crenatum in high - producing Arg strain,the ArgD - code gene of ACOAT was achived. The gene was 1176 bp in full length,encoding 390 amino acid residues. At last,all of them were expressed in C. Crenatum SYPA and Escherichia coli BL21( DE3). By selecting NI column to do the research, we had got recombinant protein whose enzyme activity was as high as 108. 2U/g after affinity purification. And then we had study its enzymatic properties. At last, recombinant Corynebacterium crenatum had been construct in order to

  8. 超声波作用下提取黑芝麻色素及稳定性试验%Study on the extraction and stability of sesame black pigment by ultrasonic wave

    Institute of Scientific and Technical Information of China (English)

    陈小全; 仇玉芹; 黄田; 邵辉莹; 翟虎; 周秀艳; 毕玉水

    2009-01-01

    采用有机溶剂浸泡法和超声波辅助技术从黑芝麻中提取色素,对不同温度、不同时间下的色素提取率进行测试,以确定最佳提取条件,比较两者的提取效率.结果表明:超声波作用下提取黑米色素能够缩短提取时间、降低提取温度.最佳提取工艺条件:提取剂:蒸馏水;料液比(g/mL):1 ∶ 20;提取温度:75℃;提取时间:30min;超声波功率:80W;色素在酸性条件下相对稳定;光照、加热、金属离子对色素的影响不大;其抗氧化性较差.

  9. 钝齿棒杆菌argR基因缺失株构建及其缺失对精氨酸生物合成途径相关基因转录水平的影响%Construction of Corynebacterium crenatum AS 1.542△argR and analysis of transcriptional levels of the related genes of arginine biosynthetic pathway

    Institute of Scientific and Technical Information of China (English)

    陈雪岚; 汤立; 焦海涛; 徐峰; 熊勇华

    2013-01-01

    [Objective] ArgR, coded by the argR gene from Corynebacterium crenatum AS 1. 542, acts as a negative regulator in arginine biosynthetic pathway. However, the effect of argR on transcriptional levels of the related biosynthetic genes has not been reported. Here, we constructed a deletion mutant of argR gene; C. Crenatum AS 1.542 △argR using marker-less knockout technology, and compared the changes of transcriptional levels of the arginine biosynthetic genes between the mutant strain and the wild-type strain. [Methods] We used marker-less knockout technology to construct C. Crenatum AS 1. 542△argR and analyzed the changes of the relate genes at the transcriptional level using real-time fluorescence quantitative PCR. [Results] C. Crenatum AS 1. 542△argR was successfully obtained and the transcriptional level of arginine biosynthetic genes in this mutant increased significantly with an average of about 162. 1 folds. [Conclusion] The arginine biosynthetic genes in C. Crenatum are clearly controlled by the negative regulator ArgR. However, the deletion of this regulator does not result in a clear change in arginine production in the bacteria.%[目的]钝齿棒杆菌AS 1.542中argR基因编码的蛋白ArgR在精氨酸生物合成途径中扮演负调控的角色,但其对相关基因在转录水平的影响还未见报道.因此,本课题组构建了钝齿棒杆菌argR基因缺失株,并在转录水平上比较野生株与缺失株精氨酸生物合成途径相关基因的变化.[方法]采用无痕敲除的方法构建了钝齿棒杆菌argR基因缺失株,并采用荧光定量PCR方法分析缺失株和野生株精氨酸生物合成途径相关基因在转录水平的变化.[结果]利用pK18mobsacB质粒中蔗糖致死基因sacB反向筛选标记及PCR方法成功筛选到钝齿棒杆菌argR基因缺失株;荧光定量PCR结果表明,argR基因缺失株精氨酸生物合成途径中相关基因在转录水平获得大量提高,平均约上调162.13倍.[结论]

  10. A preliminary study on differential proteomics of Corynebacterium crenatum in two oxygen supply models%高产精氨酸的钝齿棒杆菌在高低供氧条件下的差异蛋白质组学初步研究

    Institute of Scientific and Technical Information of China (English)

    陶帅; 窦文芳; 张晓梅; 许泓瑜; 饶志明; 许正宏

    2011-01-01

    Dissolved oxygen level directly affects the synthesis of arginine by Corynebacterium crenatum (C. crenatum). Proteomic approaches including two-dimensional electrophoresis (2-DE) and matrixassisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) were used to analyze differential expression of proteins in C. crenatum in different oxygen supply conditions. The results showed that most of the isoelectric points of the proteins on the 2-DE gels distributed from 4 to 7. Thirtytwo different protein points were detected in the pH 4-7 gels and characterized by MALDI-TOF-MS as well as peptide mass fingerprintings. Using matrixscience database, 29 protein points were identified, and they represented 27 proteins. Database research showed that the proteins performed different functions in carbohydrate metabolism, amino acid metabolism and energy metabolism. Most of the differentially expressed proteins concentrated in the pentose phosphate pathway, TCA cycle and the urea cycle. The results showed that in the high oxygen supply model, energy metabolism was significantly increased, and the oxidative stress was balanced because of the increase of pentose phosphate pathway. Meanwhile, the activation of TCA cycle and urea cycle were beneficial to the synthesis of arginine.%溶氧水平直接影响钝齿棒杆菌合成精氨酸产量的高低,运用双向电泳结合MALDI-TOF-MS及MS/MS质谱鉴定技术的方法对高低供氧条件下高产精氨酸的钝齿棒杆菌的胞内可溶性蛋白进行分离鉴定.结果表明,钝齿棒杆菌菌体蛋白的等电点主要分布在4-7的范围内,采用pH4-7的IPG胶条进行分离,得到了645±12个蛋白点.通过软件比对分析发现,在高低两种供氧条件下存在32个显著差异的蛋白点,成功鉴定出了其中的29个蛋白点,代表了27种差异蛋白.采用数据库检索发现这些蛋白参与不同的代谢途径,包括糖类代谢、氨基酸代谢及能量代谢等.其主要集中

  11. "Cara inchada" of cattle, an infectious, apparently soil antibiotics-dependant periodontitis in Brazil "Cara inchada" dos bovinos, uma periodontite infecciosa, aparentemente desencadeada por antibióticos do solo

    Directory of Open Access Journals (Sweden)

    Jürgen Döbereiner

    2000-06-01

    Full Text Available The objective of this review on the investigation of "cara inchada" in cattle (CI, pursued over the last 30 years, was to elucidate the pathogenicity of the disease and come to proper conclusions on its etiology. CI has been widely considered to be of nutritional origin, caused primarily by mineral deficiency or imbalance. However, the disease consists of a rapidly progressive periodontitis, affecting the periodontal tissues at the level of the premolars and molars during the period of tooth eruption generally starting in young calves. The disease led to great economic losses for farmers in central-western Brazil, after the occupation of new land for cattle raising in the 1960s and 1970s. The lateral enlargement of the maxillary bones of affected calves gave the disease the popular name of "cara inchada", i.e., swollen or enlarged face. The enlargement was found to be due to a chronic ossifying periostitis resulting from the purulent alveolitis of CI. Black-pigmented non-saccharolytic Bacteroides melaninogenicus, always together with Actinomyces (Corynebacterium pyogenes, were isolated in large numbers from the periodontal lesions. B. melaninogenicus could be isolated in small numbers also from the marginal gingiva of a few healthy calves maintained on CI-free farms. "In vitro"-assays showed that streptomycin and actinomycin, as well as the supernatants of cultivates of actinomycetes from soils of CI-prone farms, applied in subinhibitory concentrations to the bacteria tested, enhanced significantly (up to 10 times the adherence of the black-pigmented B.melaninogenicus to epithelial cells of the bovine gingiva. The antibiotics are apparently produced in large quantities by the increased number of soil actinomycetes, including the genus Streptomyces, that develop when soil microflora are modified by cultivating virgin forest or "Cerrado" (tree-savanna for the first time for cattle grazing. The epidemiology of CI now provides strong evidence that

  12. Comprehensive discovery and characterization of small RNAs in Corynebacterium glutamicum ATCC 13032

    OpenAIRE

    Mentz, Almut; Neshat, Armin; Pfeifer, Katharina; Pühler, Alfred; Rückert, Christian; Kalinowski, Jörn

    2013-01-01

    Background Recent discoveries on bacterial transcriptomes gave evidence that small RNAs (sRNAs) have important regulatory roles in prokaryotic cells. Modern high-throughput sequencing approaches (RNA-Seq) enable the most detailed view on transcriptomes offering an unmatched comprehensiveness and single-base resolution. Whole transcriptome data obtained by RNA-Seq can be used to detect and characterize all transcript species, including small RNAs. Here, we describe an RNA-Seq approach for comp...

  13. Importance of NADPH supply for improved L-valine formation in Corynebacterium glutamicum.

    Science.gov (United States)

    Bartek, Tobias; Blombach, Bastian; Zönnchen, Enrico; Makus, Pia; Lang, Siegmund; Eikmanns, Bernhard J; Oldiges, Marco

    2010-01-01

    Cofactor recycling is known to be crucial for amino acid synthesis. Hence, cofactor supply was now analyzed for L-valine to identify new targets for an improvement of production. The central carbon metabolism was analyzed by stoichiometric modeling to estimate the influence of cofactors and to quantify the theoretical yield of L-valine on glucose. Three different optimal routes for L-valine biosynthesis were identified by elementary mode (EM) analysis. The modes differed mainly in the manner of NADPH regeneration, substantiating that the cofactor supply may be crucial for efficient L-valine production. Although the isocitrate dehydrogenase as an NADPH source within the tricarboxylic acid cycle only enables an L-valine yield of Y(Val/Glc) = 0.5 mol L-valine/mol glucose (mol Val/mol Glc), the pentose phosphate pathway seems to be the most promising NADPH source. Based on the theoretical calculation of EMs, the gene encoding phosphoglucoisomerase (PGI) was deleted to achieve this EM with a theoretical yield Y(Val/Glc) = 0.86 mol Val/mol Glc during the production phase. The intracellular NADPH concentration was significantly increased in the PGI-deficient mutant. L-Valine yield increased from 0.49 +/- 0.13 to 0.67 +/- 0.03 mol Val/mol Glc, and, concomitantly, the formation of by-products such as pyruvate was reduced.

  14. Exploring the potential of sigma factors for strain development in Corynebacterium glutamicum

    OpenAIRE

    Taniguchi, Hironori

    2016-01-01

    A sigma factor is one of the components of RNA polymerase holoenzyme and responsible for promoter recognition and transcription initiation. Bacteria usually possess multiple genes encoding sigma factors and strictly control their activity. Activation of a specific sigma factor alters the promoter recognition of the RNA polymerase holoenzyme and enhances the transcription of a specific set of genes with similar promoter sequences. Therefore, sigma factors can be used for activating expression ...

  15. Corynebacterium glucuronolyticum causing genitourinary tract infection: Case report and review of the literature

    Directory of Open Access Journals (Sweden)

    G. Gherardi

    2015-01-01

    In this report, we describe a urethritis case caused by C. glucuronolyticum in a 37-year-old, apparently healthy male, who complained mild pain in the lower abdomen, with several urinary symptoms. While urethral and semen specimens did not yield positive results for microbiological evaluation, cultures of urine samples revealed the monomicrobial growth on blood-containing media of tiny colonies after 24 h of incubation, clearly evident only after 48 h of incubation under CO2-enriched atmosphere. Colonies were identified as C. glucuronolyticum both by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF and 16S rRNA gene sequencing. Oral ciprofloxacin gradually led to clinical improvement and, finally, to a complete recovery, in accordance with microbiological findings. In spite of its infrequent detection, C. glucuronolyticum might be a potential urogenital pathogen in males more commonly that what believed, perhaps due to slow growth leading to underrecognition; we suggest therefore to consider the organism in the differential diagnostics of bacterial diseases of the urinary tract.

  16. Evaluation of ERIC-PCR as Genotyping Method for Corynebacterium pseudotuberculosis Isolates

    Science.gov (United States)

    Dorneles, Elaine M. S.; Santana, Jordana A.; Ribeiro, Dayana; Dorella, Fernanda Alves; Guimarães, Alessandro S.; Moawad, Mohamed S.; Selim, Salah A.; Garaldi, Ana Luiza M.; Miyoshi, Anderson; Ribeiro, Márcio G.; Gouveia, Aurora M. G.; Azevedo, Vasco; Heinemann, Marcos B.; Lage, Andrey P.

    2014-01-01

    The aim of this study was to evaluate the Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) as a tool for molecular typing of C. pseudotuberculosis isolates from eight different hosts in twelve countries. Ninety-nine C. pseudotuberculosis field strains, one type strain (ATCC 19410T) and one vaccine strain (1002) were fingerprinted using the ERIC-1R and ERIC-2 primers, and the ERIC-1R+ERIC-2 primer pair. Twenty-nine different genotypes were generated by ERIC 1-PCR, 28 by ERIC 2-PCR and 35 by ERIC 1+2-PCR. The discriminatory index calculated for ERIC 1, ERIC 2, and ERIC 1+2-PCR was 0.89, 0.86, and 0.92, respectively. Epidemiological concordance was established for all ERIC-PCR assays. ERIC 1+2-PCR was defined as the best method based on suitability of the amplification patterns and discriminatory index. Minimal spanning tree for ERIC 1+2-PCR revealed three major clonal complexes and clustering around nitrate-positive (biovar Equi) and nitrate-negative (biovar Ovis) strains. Therefore, ERIC 1+2-PCR proved to be the best technique evaluated in this study for genotyping C. pseudotuberculosis strains, due to its usefulness for molecular epidemiology investigations. PMID:24901343

  17. Coordinated regulation of biosynthetic and regulatory genes coincides with anthocyanin accumulation in developing eggplant fruit

    Science.gov (United States)

    Violet to black pigmentation of eggplant (Solanum melongena) fruit is attributed to anthocyanin accumulation. Model systems support the interaction of biosynthetic and regulatory genes for anthocyanin biosynthesis. Anthocyanin structural gene transcription requires the expression of at least one m...

  18. Detection of DOPA-Melanin in the Dimorphic Fungal Pathogen Penicillium marneffei and Its Effect on Macrophage Phagocytosis In Vitro

    OpenAIRE

    Donghua Liu; Lili Wei; Ting Guo; Weifen Tan

    2014-01-01

    The fungal pathogen Penicillium marneffei produces melanin-like pigment in vitro. The synthetic pathway of melanin and its possible influence in the protective yeast cells surviving within macrophage cells are not known. In this work, P. marneffei produced brown black pigment in the presence of L-DOPA and black particles were extracted from yeast cells treated with proteolytic enzymes, denaturant and concentrated hot acid. Kojic acid inhibited the brown-black pigment production of P. marneffe...

  19. Removal of black stains from teeth by photodynamic therapy: clinical and microbiological analysis.

    Science.gov (United States)

    Pessoa, Larissa; Galvão, Virgilio; Damante, Carla; Sant'Ana, Adriana Campos Passanezi

    2015-01-01

    Black-pigmented bacteria (BPB) are Gram-negative anaerobic, non-motile, proteolytic rods strongly implicated in the pathogenesis of periodontal disease. Although pigments are produced in vitro, black pigmentation is rarely found clinically. However, it may compromise aesthetics and contribute to gingival inflammation. The aim of this report is to describe a clinical case of a 10-year-old boy showing black pigmentation covering all teeth and to propose an alternative therapy for removal of black pigmentation, based on photodynamic therapy (PDT). In order to perform microbiological analysis, plaque samples were collected before and after PDT, and analysed by real-time-PCR (RT-PCR). The results showed a significant reduction in BPB levels after therapy, along with clinical evidence of absence of black pigmentation and reduction in gingival bleeding, although the plaque index remained unaltered. This case showed that PDT is effective for eliminating black pigmentation caused by BPB, without recurrence during a follow-up period of 7 months. PMID:26701991

  20. Comparative 13C Metabolic Flux Analysis of Pyruvate Dehydrogenase Complex-Deficient, l-Valine-Producing Corynebacterium glutamicum▿†

    OpenAIRE

    Bartek, Tobias; Blombach, Bastian; Lang, Siegmund; Eikmanns, Bernhard J.; Wiechert, Wolfgang; Oldiges, Marco; Nöh, Katharina; Noack, Stephan

    2011-01-01

    l-Valine can be formed successfully using C. glutamicum strains missing an active pyruvate dehydrogenase enzyme complex (PDHC). Wild-type C. glutamicum and four PDHC-deficient strains were compared by 13C metabolic flux analysis, especially focusing on the split ratio between glycolysis and the pentose phosphate pathway (PPP). Compared to the wild type, showing a carbon flux of 69% ± 14% through the PPP, a strong increase in the PPP flux was observed in PDHC-deficient strains with a maximum o...

  1. Optimization of the IPP precursor supply for the production of lycopene, decaprenoxanthin and astaxanthin by Corynebacterium glutamicum

    Directory of Open Access Journals (Sweden)

    Sabine A.E. Heider

    2014-08-01

    Full Text Available The biotechnologically relevant bacterium C. glutamicum, currently used for the million ton-scale production of amino acids for the food and feed industries, is pigmented due to synthesis of the rare cyclic C50 carotenoid decaprenoxanthin and its glucosides. The precursors of carotenoid biosynthesis, isopenthenyl pyrophosphate (IPP and its isomer dimethylallyl pyrophosphate (DMAPP, are synthesized in this organism via the methylerythritol phosphate (MEP or non-mevalonate pathway. Terminal pathway engineering in recombinant C. glutamicum permitted the production of various nonnative C50 and C40 carotenoids. Here, the role of engineering isoprenoid precursor supply for lycopene production by C. glutamicum was characterized. Overexpression of dxs encoding the enzyme that catalyzes the first committed step of the MEP-pathway by chromosomal promoter exchange in a prophage-cured, genome-reduced C. glutamicum strain improved lycopene formation. Similarly, an increased IPP supply was achieved by chromosomal integration of two artificial operons comprising MEP pathway genes under the control of a constitutive promoter. Combined overexpression of dxs and the other six MEP pathways genes in C. glutamicum strain LYC3-MEP was not synergistic with respect to improving lycopene accumulation. Based on C. glutamicum strain LYC3-MEP astaxanthin could be produced in the mg per g cell dry weight range when the endogenous genes crtE, crtB and crtI for conversion of geranylgeranyl pyrophosphate to lycopene were coexpressed with the genes for lycopene cyclase and β-carotene hydroxylase from Pantoea ananatis and carotene C(4 oxygenase from Brevundimonas aurantiaca.

  2. Comparative 13C metabolic flux analysis of pyruvate dehydrogenase complex-deficient, L-valine-producing Corynebacterium glutamicum.

    Science.gov (United States)

    Bartek, Tobias; Blombach, Bastian; Lang, Siegmund; Eikmanns, Bernhard J; Wiechert, Wolfgang; Oldiges, Marco; Nöh, Katharina; Noack, Stephan

    2011-09-01

    L-Valine can be formed successfully using C. glutamicum strains missing an active pyruvate dehydrogenase enzyme complex (PDHC). Wild-type C. glutamicum and four PDHC-deficient strains were compared by (13)C metabolic flux analysis, especially focusing on the split ratio between glycolysis and the pentose phosphate pathway (PPP). Compared to the wild type, showing a carbon flux of 69% ± 14% through the PPP, a strong increase in the PPP flux was observed in PDHC-deficient strains with a maximum of 113% ± 22%. The shift in the split ratio can be explained by an increased demand of NADPH for l-valine formation. In accordance, the introduction of the Escherichia coli transhydrogenase PntAB, catalyzing the reversible conversion of NADH to NADPH, into an L-valine-producing C. glutamicum strain caused the PPP flux to decrease to 57% ± 6%, which is below the wild-type split ratio. Hence, transhydrogenase activity offers an alternative perspective for sufficient NADPH supply, which is relevant for most amino acid production systems. Moreover, as demonstrated for L-valine, this bypass leads to a significant increase of product yield due to a concurrent reduction in carbon dioxide formation via the PPP.

  3. Exploring the role of sigma factor gene expression on production by Corynebacterium glutamicum: sigma factor H and FMN as example

    Directory of Open Access Journals (Sweden)

    Hironori eTaniguchi

    2015-07-01

    Full Text Available 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 C. glutamicum WT was assayed using an IPTG inducible plasmid system and different IPTG concentrations. It was revealed that growth was severely decreased when sigD or sigH were overexpressed with IPTG concentrations higher than 50 μM. Overexpression of sigH led to an obvious phenotypic change, a yellow-colored supernatant. HPLC analysis revealed that riboflavin was excreted to the medium when sigH was overexpressed and DNA microarray analysis confirmed increased expression of riboflavin biosynthesis genes. In addition, genes for enzymes related to the pentose phosphate pathway and for enzymes dependent on FMN, FAD or NADPH as cofactor were upregulated when sigH was overexpressed. To test if sigH overexpression can be exploited for production of riboflavin-derived FMN or FAD, the endogenous gene for bifunctional riboflavin kinase/FMN adenyltransferase was co-expressed with sigH from a plasmid. Balanced expression of sigH and ribF improved accumulation of riboflavin (19.8 ± 0.3 μM and allowed for its conversion to FMN (33.1 ± 1.8 μM in the supernatant. While a proof-of-concept was reached, conversion was not complete and titers were not high. This study revealed that inducible and gradable overexpression of sigma factor genes is an interesting approach to switch gene expression profiles and to discover untapped potential of bacteria for chemical production.

  4. High-resolution detection of DNA binding sites of the global transcriptional regulator GlxR in Corynebacterium glutamicum

    DEFF Research Database (Denmark)

    Jungwirth, Britta; Sala, Claudia; Kohl, Thomas A;

    2013-01-01

    mapping of these data on the genome sequence of C. glutamicum, 107 enriched DNA fragments were detected from cells grown with glucose as carbon source. GlxR binding sites were identified in the sequence of 79 enriched DNA fragments, of which 21 sites were not previously reported. Electrophoretic mobility...... of the 6C non-coding RNA gene and to non-canonical DNA binding sites within protein-coding regions. The present study underlines the dynamics within the GlxR regulon by identifying in vivo targets during growth on glucose and contributes to the expansion of knowledge of this important transcriptional...

  5. Increased expression of pyruvate carboxylase and biotin protein ligase increases lysine production in a biotin prototrophic Corynebacterium glutamicum strain

    DEFF Research Database (Denmark)

    Wang, Zhihao; Moslehi-Jenabian, Soloomeh; Solem, Christian;

    2015-01-01

    was cultivated without biotin, indicating a suboptimal intracellular concentration of biotin. In an attempt to locate the potential bottleneck, we added pimelic acid, an early biotin precursor, and found that growth rate could be restored fully, which demonstrates that the bottleneck is in pimeloyl-CoA (or...... pimeloyl-Acyl Carrier Protein [ACP]) formation. Pyruvate carboxylase (pycA), a biotin-dependent enzyme needed for lysine biosynthesis and biotin ligase (birA), which is responsible for attaching biotin to pyruvate carboxylase, were overexpressed by replacing the native promoters with the strong superoxide...

  6. 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.

    Science.gov (United States)

    Wendisch, Volker F; Brito, Luciana Fernandes; Gil Lopez, Marina; Hennig, Guido; Pfeifenschneider, Johannes; Sgobba, Elvira; Veldmann, Kareen H

    2016-09-20

    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, pseudomonads, bacilli and Baker's yeast used in these large scale processes have been engineered for efficient utilization of alternative carbon sources. This flexible feedstock concept is central to the use of non-glucose second and third generation feedstocks in the emerging bioeconomy. The metabolic engineering efforts to broaden the substrate scope of E. coli, C. glutamicum, pseudomonads, B. subtilis and yeasts to include non-native carbon sources will be reviewed. Strategies to enable simultaneous consumption of mixtures of native and non-native carbon sources present in biomass hydrolysates will be summarized and a perspective on how to further increase feedstock flexibility for the realization of biorefinery processes will be given. PMID:27491712

  7. Continuous recovery of valine in a model mixture of amino acids and salt from Corynebacterium bacteria fermentation using a simulated moving bed chromatography.

    Science.gov (United States)

    Park, Chanhun; Nam, Hee-Geun; Jo, Se-Hee; Wang, Nien-Hwa Linda; Mun, Sungyong

    2016-02-26

    The economical efficiency of valine production in related industries is largely affected by the performance of a valine separation process, in which valine is to be separated from leucine, alanine, and ammonium sulfate. Such separation is currently handled by a batch-mode hybrid process based on ion-exchange and crystallization schemes. To make a substantial improvement in the economical efficiency of an industrial valine production, such a batch-mode process based on two different separation schemes needs to be converted into a continuous-mode separation process based on a single separation scheme. To address this issue, a simulated moving bed (SMB) technology was applied in this study to the development of a continuous-mode valine-separation chromatographic process with uniformity in adsorbent and liquid phases. It was first found that a Chromalite-PCG600C resin could be eligible for the adsorbent of such process, particularly in an industrial scale. The intrinsic parameters of each component on the Chromalite-PCG600C adsorbent were determined and then utilized in selecting a proper set of configurations for SMB units, columns, and ports, under which the SMB operating parameters were optimized with a genetic algorithm. Finally, the optimized SMB based on the selected configurations was tested experimentally, which confirmed its effectiveness in continuous separation of valine from leucine, alanine, ammonium sulfate with high purity, high yield, high throughput, and high valine product concentration. It is thus expected that the developed SMB process in this study will be able to serve as one of the trustworthy ways of improving the economical efficiency of an industrial valine production process.

  8. In Vitro Activities of Daptomycin, Vancomycin, Quinupristin- Dalfopristin, Linezolid, and Five Other Antimicrobials against 307 Gram-Positive Anaerobic and 31 Corynebacterium Clinical Isolates

    OpenAIRE

    Goldstein, Ellie J. C.; Citron, Diane M.; Merriam, C. Vreni; Warren, Yumi A.; Tyrrell, Kerrin L.; Fernandez, Helen T.

    2003-01-01

    The activities of daptomycin, a cyclic lipopeptide, and eight other agents were determined against 338 strains of gram-positive anaerobic bacteria and corynebacteria by the NCCLS reference agar dilution method with supplemented brucella agar for the anaerobes and Mueller-Hinton agar for the corynebacteria. The daptomycin MICs determined on Ca2+-supplemented (50 mg/liter) brucella agar plates were one- to fourfold lower than those determined in unsupplemented media. Daptomycin was highly activ...

  9. Functional genomics of pH homeostasis in Corynebacterium glutamicum revealed novel links between pH response, oxidative stress, iron homeostasis and methionine synthesis

    Directory of Open Access Journals (Sweden)

    Persicke Marcus

    2009-12-01

    Full Text Available Abstract Background The maintenance of internal pH in bacterial cells is challenged by natural stress conditions, during host infection or in biotechnological production processes. Comprehensive transcriptomic and proteomic analyses has been conducted in several bacterial model systems, yet questions remain as to the mechanisms of pH homeostasis. Results Here we present the comprehensive analysis of pH homeostasis in C. glutamicum, a bacterium of industrial importance. At pH values between 6 and 9 effective maintenance of the internal pH at 7.5 ± 0.5 pH units was found. By DNA microarray analyses differential mRNA patterns were identified. The expression profiles were validated and extended by 1D-LC-ESI-MS/MS based quantification of soluble and membrane proteins. Regulators involved were identified and thereby participation of numerous signaling modules in pH response was found. The functional analysis revealed for the first time the occurrence of oxidative stress in C. glutamicum cells at neutral and low pH conditions accompanied by activation of the iron starvation response. Intracellular metabolite pool analysis unraveled inhibition of the TCA and other pathways at low pH. Methionine and cysteine synthesis were found to be activated via the McbR regulator, cysteine accumulation was observed and addition of cysteine was shown to be toxic under acidic conditions. Conclusions Novel limitations for C. glutamicum at non-optimal pH values were identified by a comprehensive analysis on the level of the transcriptome, proteome, and metabolome indicating a functional link between pH acclimatization, oxidative stress, iron homeostasis, and metabolic alterations. The results offer new insights into bacterial stress physiology and new starting points for bacterial strain design or pathogen defense.

  10. Distribution of PLD and FagA, B, C and D genes in Corynebacterium pseudotuberculosis isolates from sheep and goats with caseus lymphadenitis

    Directory of Open Access Journals (Sweden)

    Maria da Conceição Aquino de Sá

    2013-01-01

    Full Text Available Caseous lymphadenits (CL is a chronic and subclinical disease that affects goats and sheep and, consequently, causes economic losses, especially to small producers. The purpose of this study, through use of Polymerase Chain Reaction (PCR, was to verify the presence of virulence genes of phospholipase D (PLD, integral membrane protein (FagA, iron enterobactin transporter (FagB, ATP binding cytoplasmic membrane protein (FagC and iron siderophore binding protein (FagD in 168 isolates of C. pseudotuberculosis obtained from cases of caseous lymphadenitis in goats and sheep. FagA, FagB and PLD genes were detected in all 145 strains isolated from abscesses in superficial lymph nodes and in 23 strains isolated from viscera. The FagC gene was positive in 167 (99.40% isolates. The FagD gene was detected in 160 (95.23% isolates. All virulence factors analyzed were found more frequently among isolates collected in the viscera of animals with CL, indicating a multifactorial nature, as well as variations, in the invasive potential of C. pseudotuberculosis strains.

  11. The pan-genome of the animal pathogen Corynebacterium pseudotuberculosis reveals differences in genome plasticity between the biovar ovis and equi strains

    DEFF Research Database (Denmark)

    Soares, Siomar C; Silva, Artur; Trost, Eva;

    2013-01-01

    of complete pilus structures in biovar ovis could be responsible for a remarkable ability of these strains to spread throughout host tissues and penetrate cells to live intracellularly, in contrast with the biovar equi, which rarely attacks visceral organs. Intracellularly, the biovar ovis strains...

  12. 76 FR 9049 - Notice of Intent To Repatriate Cultural Items: University of Pennsylvania Museum of Archaeology...

    Science.gov (United States)

    2011-02-16

    ... Raven of the Roof (NA10511). It is carved and painted with blue-green, red, black and white pigment, and... wolf, and is painted with green, red, black, and white pigment. Natural wolf fur, ears and teeth make.... The helmet is painted with blue-green, red, and black pigment and it is decorated with copper, and...

  13. MATERIAL AND ENVIRONMENTAL SUSTAINABILITY IN CERAMIC PROCESSING

    Science.gov (United States)

    Materials Characterization The density of 3124 Ferro Frit and Mason Chrome Free Black Pigment was measured to be 2.4911 g/cm3 and 5.0703 g/cm3 respectively. The single point, BET and Langmuir surface area of 3124 Ferro Frit was deter...

  14. Visualizing and simulating flow conditions in concrete form filling using pigments

    DEFF Research Database (Denmark)

    Jacobsen, Stefan; Cepuritis, Rolands; Peng, Ya;

    2013-01-01

    Flow variation at surfaces and reinforcement during form filling was visualized with grey and black SCC. The border between grey and black (pigmented) SCC was captured as frozen images on hardened sawn- and formwork surfaces in a flow box experiment. Maximum velocity occurred at the centre of the...

  15. The MC1R gene in the guppy (Poecilia reticulata: Genotypic and phenotypic polymorphisms

    Directory of Open Access Journals (Sweden)

    Yokoyama Jun

    2011-02-01

    Full Text Available Abstract Background The guppy (Poecilia reticulata is an important model organism for studying sexual selection; male guppies have complex and conspicuous pigmentation, and female guppies exhibit preferences for males with specific color spots. Understanding the genetic basis underlying pigmentation variation in the guppy is important for exploring the factors causing the maintenance of color polymorphism in wild populations. Findings We focused on the melanic black pigmentation of guppies, and examined genetic variations in the melanocortin 1 receptor (MC1R gene because variation in this gene is known to contribute to polymorphism of melanin pigmentation in several animal species. The complete coding sequence of the guppy MC1R gene was determined, and two different MC1R alleles (963 and 969 bp were found in wild populations. Ornamental strain guppies with a 963-bp MC1R tended to show less black pigmentation than those with a 969-bp MC1R, although the association between MC1R genotype and black pigmentation disappeared in the F2 offspring. Conclusions The guppy MC1R gene showed variation in the five wild Trinidadian populations we examined, and these populations also differed in terms of allele frequencies. We identified a significant association between black pigmentation and MC1R genotype in fish obtained from aquarium shops. However, the results from F2 families suggest that there are other genes that modify the effects of the MC1R gene.

  16. Characterization and Photoprotector Activity of Endophytic Fungal Pigments from Coastal Plant Sarang Semut (Hydnophytum formicarum

    Directory of Open Access Journals (Sweden)

    Mada Triandala Sibero

    2016-04-01

    Full Text Available Endophytic fungus RS3 isolated from coastal plant sarang semut (Hydnophytum formicarum produced extracellular black pigment. The aims of this research were to obtain the pigment, to characterize and to determine the photoprotector activity. This research was conducted into several steps, that were determination of the best precipitating agent, characterization using instrument and solubility analysis, and analysis of Sun Protection Factor (SPF. Results showed the pigment was precipitated using acid solvent with pH ≤ 2.5. Functional groups of pigment were hydroxyl, aromatic ring, phenol and amine. According to its characteristics, black pigment produced by RS3 isolate was proposed as melanin. The photoprotector analysis showed the SPF value was 11.33.

  17. Characterization and Photoprotector Activity of Endophytic Fungal Pigments from Coatal Plant Sarang Semut (Hydnophytum formicarum

    Directory of Open Access Journals (Sweden)

    Mada Triandala Sabero

    2016-04-01

    Full Text Available Isolate endophytic fungal RS3 from smooth ant plants (Hydnophytum formicarum produced black pigment. The aims of this research were to obtain the pigment, to characterize and to determine the photoprotector activity. This research was consisted into several steps, there were determined the best precipitating agent, characterization using instrument and solubility analysis, and analysis of Sun Protection Factor (SPF. Results showed the pigment was precipitated using acid solvent with pH ≤ 2,5. Functional groups of pigment pellet were hydroxy, aromatic ring, phenol and amine. According to characteristic, black pigment produced by fungal RS3 proposed as melanin. The photoprotector analysis showed SPF the value was 11.33.

  18. Fourier-transform Raman spectroscopic study of pigments in native American Indian rock art: Seminole Canyon

    Science.gov (United States)

    Edwards, H. G. M.; Drummond, L.; Russ, J.

    1998-10-01

    Samples of rock art (ca. 3000-4200 years BP) from the Lower Pecos region of Texas, near the confluences of the Pecos and Devils rivers with the Rio Grande, have been analysed using Raman microscopy. This rock art represents some of the finest pictographs known in North America. The red pigment is confirmed to be red ochre (iron (III) oxide and clay) whereas the black pigment is manganese (IV) oxide. White areas of the paintings are identified as calcium oxalate monohydrate (whewellite), whose presence could indicate the previous colonisation of the shelter walls by lichens. The black pigmented areas only contained Raman spectroscopic evidence for organic matter which was probably used as a binding agent.

  19. Oral pigmentation: A review.

    Science.gov (United States)

    Sreeja, C; Ramakrishnan, K; Vijayalakshmi, D; Devi, M; Aesha, I; Vijayabanu, B

    2015-08-01

    Pigmentations are commonly found in the mouth. They represent in various clinical patterns that can range from just physiologic changes to oral manifestations of systemic diseases and malignancies. Color changes in the oral mucosa can be attributed to the deposition of either endogenous or exogenous pigments as a result of various mucosal diseases. The various pigmentations can be in the form of blue/purple vascular lesions, brown melanotic lesions, brown heme-associated lesions, gray/black pigmentations. PMID:26538887

  20. Non-conventional therapeutics for oral infections

    OpenAIRE

    Robert P. Allaker; Ian Douglas, CW

    2015-01-01

    As our knowledge of host-microbial interactions within the oral cavity increases, future treatments are likely to be more targeted. For example, efforts to target a single species or key virulence factors that they produce, while maintaining the natural balance of the resident oral microbiota that acts to modulate the host immune response would be an advantage. Targeted approaches may be directed at the black-pigmented anaerobes, Porphyromonas gingivalis and Prevotella intermedia, associated ...

  1. Amalgam Contact Hypersensitivity Lesion: An Unusual Presentation-Report Of A Rare Case

    OpenAIRE

    Ramnarayan, BK; Maligi, PM; Smitha, T; Patil, US

    2014-01-01

    Amalgam or its components may cause Type IV hypersensitivity reactions on the oral mucosa. These amalgam contact hypersensitivity lesions (ACHL) present as white striae and plaques, erythematous, erosive, atrophic, or ulcerative lesions. Postinflammatory pigmentation in such lesions and pigmentation due to amalgam incorporation in the soft tissue have been reported in the literature. However, ACHL presenting primarily as a black pigmented lesion is extremely rare if not reported. The clinicia...

  2. Gene cluster involved in melanin biosynthesis of the filamentous fungus Alternaria alternata.

    OpenAIRE

    N. Kimura; Tsuge,T.

    1993-01-01

    The filamentous fungus Alternaria alternata produces melanin, a black pigment, from acetate via 1,8-dihydroxynaphthalene. To isolate a fungal gene required for melanin biosynthesis, we transformed an A. alternata Brm1- (light brown) mutant with the DNA of a wild-type strain genomic library constructed by use of a cosmid carrying the hygromycin B phosphotransferase gene. When hygromycin B-resistant transformants were screened for melanin production, 1 of 1,363 transformants appeared to regain ...

  3. Mechanism for the anti-thyroid action of minocycline

    Energy Technology Data Exchange (ETDEWEB)

    Doerge, D.R.; Divi, R.L.; Deck, J. [National Center for Toxicological Research, Jefferson, AR (United States); Taurog, A. [Univ. of Texas, Dallas, TX (United States)

    1997-01-01

    Administration of minocycline (MN), a tetracycline antibiotic, produces a black pigment in the thyroids of humans and several species of experimental animals and antithyroid effects in rodents. We have previously shown that these effects appear to be related to interactions of MN with thyroid peroxidase (TPO), the key enzyme in thyroid hormone synthesis. In the present study, the mechanisms for inhibition of TPO-catalyzed iodination and coupling reactions by MN were investigated. 37 refs., 7 figs., 3 tabs.

  4. Tattoo inks in general usage contain nanoparticles

    DEFF Research Database (Denmark)

    Høgsberg, T; Löschner, Katrin; Löf, D;

    2011-01-01

    the particle sizes in tattoo inks in general usage. Methods The particle size was measured by laser diffraction, electron microscopy and X-ray diffraction. Results The size of the pigments could be divided into three main classes. The black pigments were the smallest, the white pigments the largest...... in general usage is new and may contribute to the understanding of tattoo ink kinetics. How the body responds to NP tattoo pigments should be examined further....

  5. Porphyromonas endodontalisが産生する新規のアスパラギン酸特異的ジペプチジルペプチダーゼの解析

    OpenAIRE

    原賀, 裕; ハラガ, ヒロシ; Hiroshi, HARAGA

    2008-01-01

    Porphyromonas endodontalis, a black-pigmented anaerobe, is a predominant pathogen of human periapical periodontitis. In contrast to P. gingivalis, a major pathogen of chronic periodontitis, the pathogenic factor(s) of P. endodontalis has been poorly characterized. In this study, the protease activities in the P. endodontalis extracellular fraction were examined using various MCA polypeptides. The results indicated that the extracellular fraction had a spectrum of proteolytic activities distin...

  6. Relation of cholesterol metabolism to pediatric gallstone disease: a retrospective controlled study

    OpenAIRE

    Koivusalo, Antti; Pakarinen, Mikko; Gylling, Helena; Nissinen, Markku J

    2015-01-01

    Background Cholesterol metabolism may be involved in pediatric gallstone disease. We aimed to reveal cholesterol metabolites and phytosterols and their relation to stone composition of sterols in children having black pigment and cholesterol stones. Methods We performed retrospective controlled clinical study, in which we examined parameters of cholesterol metabolism and liver function values in serum (n = 28) and gallstones (n = 46) of consecutively cholecystectomized children. Serum values ...

  7. The macromelanophore locus and the melanoma oncogene Xmrk are separate genetic entities in the genome of Xiphophorus.

    OpenAIRE

    Weis, S; Schartl, M.

    1998-01-01

    Fish of the genus Xiphophorus are polymorphic for black pigmentation patterns. Certain intra- or interspecific hybrids exhibit enhanced expression of these patterns, leading in many cases to malignant melanoma. Because no recombination was ever observed between the pattern information and the genetic predisposition to develop melanoma after hybridization, a "tumor gene" (Tu) was postulated that encodes both phenotypes. A dominant oncogene, ONC-Xmrk, was then found to be necessary and sufficie...

  8. NCBI nr-aa BLAST: CBRC-MMUR-01-1271 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-1271 ref|ZP_03710111.1| hypothetical protein CORMATOL_00929 [Corynebacterium matruchoti...i ATCC 33806] ref|ZP_04835772.1| serine/threonine-protein kinase PknA [Corynebacterium matruchoti...i ATCC 14266] gb|EEG27500.1| hypothetical protein CORMATOL_00929 [Corynebacterium matruchotii ATCC 338...06] gb|EES80905.1| serine/threonine-protein kinase PknA [Corynebacterium matruchotii ATCC 14266] ZP_03710111.1 7.7 32% ...

  9. Sexy faces in a male paper wasp.

    Directory of Open Access Journals (Sweden)

    André Rodrigues de Souza

    Full Text Available Sexually selected signals are common in many animals, though little reported in social insects. We investigated the occurrence of male visual signals mediating the dominance relationships among males and female choice of sexual partner in the paper wasp Polistes simillimus. Males have three conspicuous, variable and sexually dimorphic traits: black pigmentation on the head, a pair of yellow abdominal spots and body size differences. By conducting behavioral assays, we found that none of the three visual traits are associated with male-male dominance relationship. However, males with higher proportion of black facial pigmentation and bigger yellow abdominal spots are more likely chosen as sexual partners. Also, after experimentally manipulating the proportion of black pigment on males' face, we found that females may evaluate male facial coloration during the choice of a sexual partner. Thus, the black pigmentation on P. simillimus male's head appears to play a role as a sexually selected visual signal. We suggest that sexual selection is a common force in Polistes and we highlight the importance of this group as a model for the study of visual communication in insects.

  10. Sexy faces in a male paper wasp.

    Science.gov (United States)

    de Souza, André Rodrigues; Alberto Mourão Júnior, Carlos; do Nascimento, Fabio Santos; Lino-Neto, José

    2014-01-01

    Sexually selected signals are common in many animals, though little reported in social insects. We investigated the occurrence of male visual signals mediating the dominance relationships among males and female choice of sexual partner in the paper wasp Polistes simillimus. Males have three conspicuous, variable and sexually dimorphic traits: black pigmentation on the head, a pair of yellow abdominal spots and body size differences. By conducting behavioral assays, we found that none of the three visual traits are associated with male-male dominance relationship. However, males with higher proportion of black facial pigmentation and bigger yellow abdominal spots are more likely chosen as sexual partners. Also, after experimentally manipulating the proportion of black pigment on males' face, we found that females may evaluate male facial coloration during the choice of a sexual partner. Thus, the black pigmentation on P. simillimus male's head appears to play a role as a sexually selected visual signal. We suggest that sexual selection is a common force in Polistes and we highlight the importance of this group as a model for the study of visual communication in insects. PMID:24849073

  11. Blackberry pigment (whitlockite) gallstones in uremic patient.

    Science.gov (United States)

    Cariati, Andrea

    2013-04-01

    Black pigment gallstones represent nearly the 15% of all gallstones and are usually related with the typical "hyperbilirubinbilia" factors as hemolysis, ineffective erythropoiesis, pathologic enterohepatic cycling of unconjugated bilirubin, cirrhosis and with gallbladder mucosa (parietal) factors as adenomyomatosis. During a prospective study on 179 patients who underwent cholecystectomy for gallstone disease a 69-year-old female with predialysis chronic kidney disease was operated for symptomatic gallstone. The removed gallstones were black pigment gallstones, with an irregular (as small blackberry) surface. Analysis of the stones revealed a great amount of whitlockite (Ca Mg)3 (PO4)2. Recent studies on chronic renal failure patients found that chronic uremia is associated with an increased risk of gallstones formation (22%) as it seems in women affected by primary hyperparathyroidism (30%). The presence of calcium phosphate gallstones in these patients have been never described. In conclusion, further studies could be necessary to establish the role of chronic renal failure and of primary and secondary hyperparathyroidism in gallstones formation and, in particular, if dialysis and predialysis patients have an higher risk to develop cholesterol and black pigment gallstones in particular of the "blackberry" (whitlockite) subtype. PMID:22959097

  12. Identification of titanium in human tissues: probable role in pathologic processes

    International Nuclear Information System (INIS)

    Six cases of titanium dioxide exposure involving lung, skin, and synovium are described, with a review of the literature. The patients, four men and two women, were between the ages of 22 and 65 years. The pulmonary changes were characterized by fibrosis and numerous macrophages with abundant deposition of a black pigment. Adjacent areas of bronchopneumonia were also observed. In the skin a severe necrotizing lesion involving the subcutaneous tissue with extension to the muscle was observed in one case and a nonspecific inflammatory response was observed in another; both cases showed abundant black pigment deposition. Electron microscopy and energy dispersive x-ray analysis demonstrated the presence of large quantities of titanium in the pigment granules. There may be a combination of black pigment deposition and fibrosis, necrosis, or a xanthomatous or granulomatous reaction, that, together with negative results on special staining and culture studies for organisms, should raise the suspicion of titanium-associated injury and prompt the study of the affected tissues by x-ray analysis for positive identification

  13. Black Thyroid Associated with Thyroid Carcinoma

    Directory of Open Access Journals (Sweden)

    Emad Kandil

    2010-01-01

    Full Text Available Objective. Black thyroid is a rare pigmented change seen almost exclusively in patients upon minocycline ingestion, and the process has previously been thought to be generally benign. There have been 61 reported cases of black thyroid. We are aware of 13 cases previously reported in association with thyroid carcinoma. This paper reports six patients with black thyroid pigmentation in association with thyroid carcinoma. Design. The medical records of six patients who were diagnosed with black thyroid syndrome, all of whom underwent thyroid surgery, were reviewed. Data on age, gender, race, preoperative fine needle aspiration biopsy (FNA, thyroid function levels, and pathology reports were collected. Main Outcome. The mean age was 60 years. There were 5 females, 4 of whom were African American. All patients were clinically and biochemically euthyroid. Black pigmentation was not diagnosed in preoperative FNA, and only one patient had a preoperative diagnosis of papillary thyroid carcinoma. The other patients underwent surgery and were found to have black pigmentation of the thyroid associated with carcinoma. Conclusions. FNA does not diagnose black thyroid, which is associated with thyroid carcinoma. Thyroid glands with black pigmentation deserve thorough pathologic examination, including several sections of each specimen.

  14. Evaluation of methodology for identification of Corynebacterium%VITEK-2 Compact仪器法鉴定棒状杆菌属细菌的方法学评价

    Institute of Scientific and Technical Information of China (English)

    杨雪静; 曹俊敏; 张伟珍

    2013-01-01

    目的 评价全自动微生物鉴定系统鉴定棒状杆菌属细菌的准确率及可靠性.方法 分别选用VITEK-2 Compact系统、API CORY系统、Vitek-32系统鉴定37株纹带棒状杆菌,5株无枝菌酸棒状杆菌,两种方法间的比较采用卡方检验,同时以“API CORY系统”为金标准,计算VITEK-2 Compact仪器法的特异性、敏感性.结果 经卡方检验,VITEK-2 Compact系统在鉴定纹带棒状杆菌方面具有很好的一致性,特异性为100%,敏感性为100%,kappa值为1.0;VITEK-2 Compact系统在鉴定无枝菌酸棒状杆菌方面一致性较差,kappa值为0.API鉴定为纹带棒状杆菌的37株菌,VITEK-32均错误的鉴定为干燥棒状杆菌,5株无枝菌酸棒状杆菌在Vitek-32鉴定系统中,生长不良.结论 VITEK-2 compact在鉴定纹带棒状杆菌方面较准确可靠,但在鉴定其他棒状杆菌方面还需进一步研究.Vitek-32系统不能准确的鉴定棒状杆菌属细菌.

  15. 纹带棒状杆菌四环素、大环内酯类抗生素相关耐药基因的研究%Research on resistance genes of corynebacterium to tetracycline and macrolide antibiotics

    Institute of Scientific and Technical Information of China (English)

    杨雪静; 侯佳惠; 曹俊敏; 王原

    2014-01-01

    目的 了解纹带棒状杆菌临床分离株的耐药状态,研究大环内酯类、四环素类抗生素耐药基因的分布情况.方法 选用API CORY系统分离纹带棒状杆菌,采用微量稀释法进行药敏试验,对ermX、tetAB基因进行PCR扩增及双向测序,后在GenBank中比对.结果 37株纹带棒状杆菌单独携带ermX基因的耐药表型为4株,占10.8%;单独携带tetAB基因的耐药表型为1株,占2.7%;同时携带ermX基因和tetAB基因的耐药表型为27株,占72.8%;不合ermX基因和tetAB基因的敏感菌有5株,占13.5%.测得序列经GenBank Blast比对,同源性达99%.结论 纹带棒状杆菌对红霉素、四环素的耐药性较高,其耐药主要与ermX基因、tetAB基因决定簇有关.

  16. Constitutive expression of citrulline biosynthetic genes in Corynebacterium glutamicum%瓜氨酸生产相关基因簇在谷氨酸棒杆菌中的组成型表达

    Institute of Scientific and Technical Information of China (English)

    谢红翠; 郝宁; 韦萍; 严明; 许琳

    2011-01-01

    构建1种组成型载体并将载体应用在表达瓜氨酸相关基因簇argCJBDF上.通过去除pXMJ19诱导型启动子上游阻遏蛋白lacⅠ基因的方法,构建组成型质粒pXMJ19 -lacⅠ,并将谷氨酸棒杆菌中合成瓜氨酸途径的基因簇argCJBDF克隆到改造过的组成型载体中,实现瓜氨酸合成相关基因簇argCJBDF在谷氨酸棒杆菌的组成型表达.结果表明:重组菌在30℃摇瓶发酵72 h后,N-乙酰谷氨酸激酶的酶活达到(0.323±0.015) U/mg,瓜氨酸的产量达到4.33 g/L.成功构建的组成型表达载体,实现了外源基因簇argCJBDF在谷氨酸棒杆菌中的组成型表达.

  17. Verification of a novel NADH-binding motif: combinatorial mutagenesis of three amino acids in the cofactor-binding pocket of Corynebacterium 2,5-diketo-D-gluconic acid reductase.

    Science.gov (United States)

    Banta, Scott; Anderson, Stephen

    2002-12-01

    A screening method has been developed to support randomized mutagenesis of amino acids in the cofactor-binding pocket of the NADPH-dependent 2,5-diketo-D-gluconic acid (2,5-DKG) reductase. Such an approach could enable the isolation of an enzyme that can better catalyze the reduction of 2,5-DKG to 2-keto-L-gulonic acid (2-KLG) using NADH as a cofactor. 2-KLG is a valuable precursor to ascorbic acid, or vitamin C, and an enzyme with increased activity with NADH may be able to improve two potential vitamin C production processes. Previously we have identified three amino acid residues that can be mutated to improve activity with NADH as a cofactor. As a pilot study to show feasibility, a library was made with these three amino acids randomized, and 300 random colonies were screened for increased NADH activity. The activities of seven mutants with apparent improvements were verified using activity-stained native gels, and sequencing showed that the amino acids obtained were similar to some of those already discovered using rational design. The four most active mutants were purified and kinetically characterized. All of the new mutations resulted in apparent kcat values that were equal to or higher than that of the best mutant obtained through rational design. At saturating levels of cofactor, the best mutant obtained was almost twice as active with NADH as a cofactor as the wild-type enzyme is with NADPH. This screen is a valuable tool for improving 2,5-DKG reductase, and it could easily be modified for improving other aspects of this protein or similar enzymes.

  18. 短小棒状杆菌菌苗治疗恶性胸腔积液临床观察%Intrapleural Corynebacterium Parvum for The Treatment of Malignant Pleural Effusion

    Institute of Scientific and Technical Information of China (English)

    周陈西; 章杰

    2003-01-01

    [目的]评价短小棒状杆菌菌苗治疗恶性胸腔积液的疗效和毒性.[方法]应用短小棒状杆菌菌苗胸腔内给药治疗46例恶性胸腔积液患者.[结果]有效率76.1%,其中完全缓解率47.8%;毒副反应主要为发热和胸痛,预防性用药后可控制.[结论]短小棒状杆菌菌苗治疗恶性胸腔积液疗效肯定,毒副反应可控制,充分引流、联合全身化疗可提高疗效.

  19. Effect of Redox Potential Regulation on Metabolic Flux Distribution of Corynebacterium crenatum%氧化还原电位调控对钝齿棒杆菌代谢通量分布的影响

    Institute of Scientific and Technical Information of China (English)

    陈小举; 操新民; 姜绍通; 李兴江

    2016-01-01

    研究不同氧化还原电位对钝齿棒杆菌厌氧发酵特性的影响,并对菌株的代谢通量分布特征进行了分析.结果表明,氧化还原电位由-56 mV降为-400 mV时,发酵液中琥珀酸质量浓度由14g/L上升为20.2 g/L,乳酸质量浓度由44.9 g/L下降为35.2 g/L.代谢通量分析结果表明,降低氧化还原电位对6-磷酸葡萄糖与磷酸烯醇式丙酮酸节点处的代谢流分布影响显著.氧化还原电位为-400 mV时,胞内戊糖磷酸途径(pentose phosphate pathway,HMP)代谢通量与-56 mV相比增加了1.74倍,由磷酸烯醇式丙酮酸流向C4途径的代谢通量与-56 mV相比增加了78%,琥珀酸通量由31.73 mmol/ (L·g·h)增加到56.53 mmol/ (L·g·h),乳酸代谢通量由159.73 mmol/ (L·g·h)下降为133.50 mtmol/ (L·g·h).研究结果表明6-磷酸葡萄糖与磷酸烯醇式丙酮酸节点是影响钝齿棒杆菌厌氧发酵产琥珀酸的关键节点,为后期通过菌种改造以调节乳酸和琥珀酸的生成比、实现乳酸与琥珀酸联产奠定了基础.

  20. NCBI nr-aa BLAST: CBRC-CREM-01-1346 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CREM-01-1346 ref|NP_939160.1| Na(+)/H(+) antiporter homolog [Corynebacterium diphtheria...e NCTC 13129] emb|CAE49312.1| Na(+)/H(+) antiporter homolog [Corynebacterium diphtheriae] NP_939160.1 2e-34 34% ...

  1. NCBI nr-aa BLAST: CBRC-AGAM-01-0038 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-AGAM-01-0038 ref|NP_938793.1| Putative cytochrome C biogenesis protein [Corynebacterium diphtheria...e NCTC 13129] emb|CAE48916.1| Putative cytochrome C biogenesis protein [Corynebacterium diphtheriae] NP_938793.1 1.6 24% ...

  2. NCBI nr-aa BLAST: CBRC-PTRO-20-0007 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PTRO-20-0007 ref|NP_938605.1| immunity-specific protein Beta241 [Corynebacteri...um diphtheriae NCTC 13129] emb|CAE48718.1| immunity-specific protein Beta241 [Corynebacterium diphtheriae] NP_938605.1 1e-04 24% ...

  3. In Vitro Activities of Ketolide HMR 3647, Macrolides, and Clindamycin against Coryneform Bacteria

    OpenAIRE

    Martínez-Martínez, Luis; Pascual, Alvaro; Suárez, Ana Isabel; Perea, Evelio J.

    1998-01-01

    The in vitro activity of ketolide HMR 3647 against coryneform bacteria isolated from clinical samples was evaluated. Except against Corynebacterium jeikeium and C. urealyticum, HMR 3647 showed high activity against Corynebacterium spp., being more active than 14- and 16-membered macrolides, azithromycin, or clindamycin. HMR 3647 also had high in vitro activity against Brevibacterium spp. and Listeria monocytogenes.

  4. NCBI nr-aa BLAST: CBRC-GGOR-01-1107 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-GGOR-01-1107 ref|ZP_03919908.1| Fe-S oxidoreductase [Corynebacterium pseudogenital...ium ATCC 33035] gb|EEJ39540.1| Fe-S oxidoreductase [Corynebacterium pseudogenitalium ATCC 33035] ZP_03919908.1 0.029 34% ...

  5. NCBI nr-aa BLAST: CBRC-MMUR-01-1316 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-1316 ref|ZP_03919908.1| Fe-S oxidoreductase [Corynebacterium pseudogenital...ium ATCC 33035] gb|EEJ39540.1| Fe-S oxidoreductase [Corynebacterium pseudogenitalium ATCC 33035] ZP_03919908.1 2e-11 34% ...

  6. T-cell-predominant lymphoid hyperplasia in a tattoo*

    OpenAIRE

    Souza, Erica Sales; Rocha, Bruno de Oliveira; Batista, Everton da Silva; de Oliveira, Rodrigo Ferreira; Farre, Lourdes; Bittencourt, Achilea Lisboa

    2014-01-01

    Cutaneous lymphoid hyperplasia (CLH) can be idiopathic or secondary to external stimuli, and is considered rare in tattoos. The infiltrate can be predominantly of B or T-cells, the latter being seldom reported in tattoos. We present a case of a predominantly T CLH, secondary to the black pigment of tattooing in a 35-year-old patient, with a dense infiltrate of small, medium and scarce large T-cells. Analysis of the rearrangement of T-cells receptor revealed a polyclonal proliferation. Since t...

  7. A new species of hydrobiid snails (Mollusca, Gastropoda, Hydrobiidae from central Greece

    Directory of Open Access Journals (Sweden)

    Canella Radea

    2011-10-01

    Full Text Available A new minute valvatiform species belonging to the genus Daphniola Radoman 1973, Daphniola eptalophos sp. n., from mountain Parnassos, Greece is described. The new species has a transparent valvatiform-planispiral shell, wide and open umbilicus, grey-black pigmented soft body and head and a black penis with a small colorless outgrowth on the left side near its base. A comparative table of shell dimensions and a key to the species known for this endemic genus for Greece are provided.

  8. Pseudomelanosis of stomach, duodenum, and jejunum.

    Science.gov (United States)

    Rustagi, Tarun; Mansoor, Muhammad S; Gibson, Joanna A; Kapadia, Cyrus R

    2015-02-01

    Pseudomelanosis is a rare finding during upper gastrointestinal endoscopy, and is most commonly seen in the duodenum. Involvement of other organs in the upper gastrointestinal tract is extremely rare, with only 1 reported case involving the stomach, duodenum, and jejunum. We present a case of a 60-year-old woman with mild anemia and hematemesis, who was found to have characteristic speckled pattern of gray-black pigmentation on endoscopic examination. To the best of our knowledge, this is the second reported case of pseudomelanosis involving the stomach, duodenum, and jejunum.

  9. Neodrassex, a new genus of the Leptodrassex group (Araneae, Gnaphosidae from South America

    Directory of Open Access Journals (Sweden)

    Ricardo Ott

    2012-09-01

    Full Text Available The new genus Neodrassex is proposed to include two new species of Gnaphosidae from Brazil. Neodrassex aureus sp. nov. is described from Amazonas, Paraná and Rio Grande do Sul states, and N. iguatemi sp. nov. is described from Paraná state. Neodrassex gen. nov. is characterized by small size, pale coloration, large anterior median eyes surrounded by black pigmentation, absence of a dorsal abdominal scutum in males and by the cheliceral dentition with 2-3 teeth on the promargin and 2-4 on the retromargin. The new genus is tentatively placed at the Leptodrassex group.

  10. A 16-Year-Old Female with Peutz-Jeghers Syndrome

    Directory of Open Access Journals (Sweden)

    Mufti Munsurar Rahman

    2014-09-01

    Full Text Available Peutz-Jeghers syndrome is a rare autosomal dominant disorder of hamartomatous polyposis of the gastrointestinal tract, with pigmentation around lips and macules on the buccal mucosa that typically manifests itself as recurrent colicky abdominal pain and intestinal obstruction due to intussusception. Here we report a case of a 16-year-old girl who presented with abdominal pain, vomiting and previous history of laparotomy for intussusception. Multiple well demarcated black pigmented macules on lips, perioral region, buccal mucosa, digits, palms and soles were noted. She was diagnosed as a case of Peutz-Jeghers syndrome and managed conservatively.

  11. Antibiotic Extraction as a Recent Biocontrol Method for Aspergillus Niger andAspergillus Flavus Fungi in Ancient Egyptian mural paintings

    Science.gov (United States)

    Hemdan, R. Elmitwalli; Fatma, Helmi M.; Rizk, Mohammed A.; Hagrassy, Abeer F.

    Biodeterioration of mural paintings by Aspergillus niger and Aspergillus flavus Fungi has been proved in different mural paintings in Egypt nowadays. Several researches have studied the effect of fungi on mural paintings, the mechanism of interaction and methods of control. But none of these researches gives us the solution without causing a side effect. In this paper, for the first time, a recent treatment by antibiotic "6 penthyl α pyrone phenol" was applied as a successful technique for elimination of Aspergillus niger and Aspergillus flavus. On the other hand, it is favorable for cleaning Surfaces of Murals executed by tembera technique from the fungi metabolism which caused a black pigments on surfaces.

  12. A portable PIXE system for the in situ characterisation of black and red pigments in neolithic, copper age and bronze age pottery

    International Nuclear Information System (INIS)

    The results of the determination of the chromophoric elements in black and red pigments in ancient neolithic, copper age and bronze age pottery are presented. The measurements were carried out by using a recently developed portable PIXE system and were performed at the Archaeological Museum of Licata (Sicily). The results confirm the presence of Manganese in black pigments of Castelluccio style pottery and indicate, for the first time, a strong presence of manganese in copper age pottery of the Serraferlicchio style, and in neolithic pottery of the Serra d'Alto style

  13. Diversity of l-Ieucine catabolism in various microorganisms involved in dairy fermentations, and identification on the rate-controlling step in the formation of the potent flavour component 3-methylbutanal.

    NARCIS (Netherlands)

    Smit, B.A.; Engels, W.J.M.; Wouters, J.T.M.; Smit, G.

    2004-01-01

    Various microorganisms, belonging to the genera Lactococcus, Lactobacillus, Streptococcus, Leuconostoc, Bifidobacterium, Propionibacterium, Brevibacterium, Corynebacterium and Arthrobacter, used in dairy fermentations such as cheese making, were analysed for their potential to convert leucine into f

  14. Coryneform bacteria in infectious diseases: clinical and laboratory aspects.

    OpenAIRE

    Coyle, M B; Lipsky, B A

    1990-01-01

    Coryneform isolates from clinical specimens frequently cannot be identified by either reference laboratories or research laboratories. Many of these organisms are skin flora that belong to a large number of taxonomic groups, only 40% of which are in the genus Corynebacterium. This review provides an update on clinical presentations, microbiological features, and pathogenic mechanisms of infections with nondiphtheria Corynebacterium species and other pleomorphic gram-positive rods. The early l...

  15. Pectinase Activity of Anaerobic and Facultatively Anaerobic Bacteria Associated with Soft Rot of Yam (Diascorea rotundata)

    OpenAIRE

    Obi, Samuel K. C.

    1981-01-01

    Anaerobic and facultatively anaerobic bacteria associated with soft rot of yam (Diascorea rotundata) were isolated by the looping-out method and found to consist of Clostridium (three isolates), Corynebacterium (three isolates), Vibrio (one isolate), and Bacillus lentus (one isolate). Enzyme assay for hydrolase, lyase, and pectinesterase activities by the cup-plate method showed that except for Vibrio sp., B. lentus, and two isolates of Corynebacterium no pectinase activity could be detected ...

  16. Culture-based identification of pigmented Porphyromonas and Prevotella species in primary endodontic infections.

    Science.gov (United States)

    Rajaram, Anuradha; Kotrashetti, Vijayalakshmi S; Somannavar, Pradeep D; Ingalagi, Preeti; Bhat, Kishore

    2016-01-01

    Background. The most common species isolated from primary endodontic infections are black-pigmented bacteria. These species are implicated in apical abscess formation due to their proteolytic activity and are fastidious in nature. Therefore, the present study was carried out to evaluate the presence and identification of various pigmented Porphyromonas and Prevotella species in the infected root canal through culture-based techniques. Methods. Thirty-one patients with primary endodontic infections were selected. Using sterile paper points, samples were collected from the root canals after access opening and prior to obturation, which were cultured using blood and kanamycin blood agar. Subsequently, biochemical test was used to identify the species and the results were analyzed using percentage comparison analysis, McNemar and chi-squared tests, Wilcoxon match pair test and paired t-test. Results. Out of 31 samples 26 were positive for black-pigmented organisms; the predominantly isolated species were Prevotella followed by Porphyromonas. In Porphyromonas only P. gingivalis was isolated. One of the interesting features was isolation of P. gingivalis through culture, which is otherwise very difficult to isolate through culture. Conclusion . The presence of Prevotella and Porphyromonas species suggests that a significant role is played by these organisms in the pathogenesis of endodontic infections. PMID:27651878

  17. The three-dimensional elemental distribution based on the surface topography by confocal 3D-XRF analysis

    Science.gov (United States)

    Yi, Longtao; Qin, Min; Wang, Kai; Lin, Xue; Peng, Shiqi; Sun, Tianxi; Liu, Zhiguo

    2016-09-01

    Confocal three-dimensional micro-X-ray fluorescence (3D-XRF) is a good surface analysis technology widely used to analyse elements and elemental distributions. However, it has rarely been applied to analyse surface topography and 3D elemental mapping in surface morphology. In this study, a surface adaptive algorithm using the progressive approximation method was designed to obtain surface topography. A series of 3D elemental mapping analyses in surface morphology were performed in laboratories to analyse painted pottery fragments from the Majiayao Culture (3300-2900 BC). To the best of our knowledge, for the first time, sample surface topography and 3D elemental mapping were simultaneously obtained. Besides, component and depth analyses were also performed using synchrotron radiation confocal 3D-XRF and tabletop confocal 3D-XRF, respectively. The depth profiles showed that the sample has a layered structure. The 3D elemental mapping showed that the red pigment, black pigment, and pottery coat contain a large amount of Fe, Mn, and Ca, respectively. From the 3D elemental mapping analyses at different depths, a 3D rendering was obtained, clearly showing the 3D distributions of the red pigment, black pigment, and pottery coat. Compared with conventional 3D scanning, this method is time-efficient for analysing 3D elemental distributions and hence especially suitable for samples with non-flat surfaces.

  18. Radiocarbon dating and compositional analysis of pre-Columbian human bones

    International Nuclear Information System (INIS)

    Analysis of ancient human bones found in “El Cóporo”, an archaeological site in Guanajuato, Mexico; were performed using a multi techniques scheme: 14C radiocarbon dating, IBA (Ion Beam Analysis), SEM-EDS (Scanning Electron Microscope Energy Dispersive X-ray Spectroscopy). We measured the elemental composition of the bones, especially some with a superficial black pigmentation. Soil samples collected from the burial place were also analyzed. The 14C dating was performed with a new High Voltage Europe 1 MV Tandentron Accelerator Mass Spectrometer (AMS) recently installed in the IFUNAM (Instituto de Física, Universidad Nacional Autónoma de México). The radiocarbon dating allowed us to determine the date of death of the individual in a period between the year 890 and 975 AD, which is consistent with the late period of the Cóporo civilization. The element sample analysis of bones with the surface black pigmentation show higher levels of Fe, Mn and Ba compared when bone’s black surface was mechanically removed. These three elements were found in soil samples from the skeleton burial place. These results indicate more likely that the bone black coloration is due to a postmortem alteration occurring in the burial environment

  19. Ternary complexes of albumin-Mn(II)-bilirubin and Electron Spin Resonance studies of gallstones

    CERN Document Server

    Chikvaidze, E N; Kirikashvili, I N; Mamniashvili, G I

    2009-01-01

    The stability of albumin-bilirubin complex was investigated depending on pH of solution. It was shown that the stability of complex increases in presence of Mn(II) ions. It was also investigated the paramagnetic composition of gallstones by the electron spin resonance (ESR) method. It turned out that all investigated gallstones contain a free bilirubin radical-the stable product of its radical oxidation. Accordingly the paramagnetic composition gallstones could be divided on three main types: cholesterol, brown pigment and black pigment stones. ESR spectra of cholesterol stones is singlet with g=2.003 and splitting between components 1.0 mT. At the same time the brown gallstones, besides aforementioned signal contain the ESR spectrum which is characteristics for Mn(II) ion complexes with inorganic compounds and, finally, in the black pigment stones it was found out Fe(III) and Cu(II) complexes with organic compounds and a singlet of bilirubin free radical. It is supposed that crystallization centers of gallst...

  20. Radiocarbon dating and compositional analysis of pre-Columbian human bones

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, E., E-mail: andrade@fisica.unam.mx [Instituto de Física, Universidad Nacional Autónoma de México, Apartado Postal 20-364, 01000 México D.F. (Mexico); Solís, C.; Canto, C.E.; Lucio, O.G. de [Instituto de Física, Universidad Nacional Autónoma de México, Apartado Postal 20-364, 01000 México D.F. (Mexico); Chavez, E. [ESIME-Z, Instituto Politécnico Nacional, ALM Zacatenco, 07738 México D.F. (Mexico); Rocha, M.F.; Villanueva, O.; Torreblanca, C.A. [Centro INAH Zacatecas, Miguel Auza No. 205, Col. Centro, Zacatecas/Zacatecas CP 98000 (Mexico)

    2014-08-01

    Analysis of ancient human bones found in “El Cóporo”, an archaeological site in Guanajuato, Mexico; were performed using a multi techniques scheme: {sup 14}C radiocarbon dating, IBA (Ion Beam Analysis), SEM-EDS (Scanning Electron Microscope Energy Dispersive X-ray Spectroscopy). We measured the elemental composition of the bones, especially some with a superficial black pigmentation. Soil samples collected from the burial place were also analyzed. The {sup 14}C dating was performed with a new High Voltage Europe 1 MV Tandentron Accelerator Mass Spectrometer (AMS) recently installed in the IFUNAM (Instituto de Física, Universidad Nacional Autónoma de México). The radiocarbon dating allowed us to determine the date of death of the individual in a period between the year 890 and 975 AD, which is consistent with the late period of the Cóporo civilization. The element sample analysis of bones with the surface black pigmentation show higher levels of Fe, Mn and Ba compared when bone’s black surface was mechanically removed. These three elements were found in soil samples from the skeleton burial place. These results indicate more likely that the bone black coloration is due to a postmortem alteration occurring in the burial environment.

  1. The macromelanophore locus and the melanoma oncogene Xmrk are separate genetic entities in the genome of Xiphophorus.

    Science.gov (United States)

    Weis, S; Schartl, M

    1998-08-01

    Fish of the genus Xiphophorus are polymorphic for black pigmentation patterns. Certain intra- or interspecific hybrids exhibit enhanced expression of these patterns, leading in many cases to malignant melanoma. Because no recombination was ever observed between the pattern information and the genetic predisposition to develop melanoma after hybridization, a "tumor gene" (Tu) was postulated that encodes both phenotypes. A dominant oncogene, ONC-Xmrk, was then found to be necessary and sufficient for the transforming function of Tu. Here we present molecular evidence that ONC-Xmrk and the pigment pattern information are encoded by separate, although intimately linked loci. No ONC-Xmrk gene was present in the 15 Xiphophorus strains investigated which exhibit no black pigmentation pattern. Five different patterns from Xiphophorus maculatus, X. evelynae, X. milleri, X. cortezi, and X. montezumae were associated with ONC-Xmrk and were melanomagenic, while fish of X. helleri, X. variatus, X. nezahualcoyotl, and X. montezumae with five other patterns had no ONC-Xmrk and consequently did not produce hybrid melanoma. These data provide evidence that ONC-Xmrk is sufficient for tumorigenesis in Xiphophorus hybrids, and that a separate, pigment pattern-encoding locus is closely linked to it. PMID:9691046

  2. 棒杆菌属细菌分类鉴定的方法学比较及对四环素与大环内酯类抗菌药物耐药机制的研究%Methodology of identification of Corynebacterium and the drug resistant mechanisms to tetracycline and macrolides antibiotics

    Institute of Scientific and Technical Information of China (English)

    曹俊敏; 杨雪静; 王原

    2013-01-01

    目的 评价全自动微生物鉴定系统鉴定棒杆菌属的准确率及可靠性;分析棒杆菌属的耐药状态,探讨其对大环内酯类、四环素类抗菌药物的主要耐药机制,为临床合理使用相关抗菌药物提供依据.方法 选用VITEK-2Compact系统、API CORY系统、VITEK-32系统对分离的37株纹带棒杆菌、5株无枝菌酸棒杆菌进行分类鉴定并比较,采用微量稀释法进行药敏试验,对ermX、tetAB基因进行PCR扩增及双向测序后,在GenBank中比对.结果 API鉴定为纹带棒杆菌的37株菌,VITEK-32均错误地鉴定为干燥棒杆菌,VITEK-2 Compact鉴定正确;37株菌对红霉素、四环素的耐药率分别为83.8%、75.7%;28株菌在染色体DNA中检测到tetAB基因,31株菌染色体DNA检测到ermX基因.结论 VITEK-2 Compact在鉴定纹带棒杆菌方面较准确可靠,但在鉴定其他棒杆菌方面有限制;纹带棒杆菌对大环内酯类、四环素的耐药率较高,染色体中获得ermX基因、tetAB基因决定簇是其耐药的主要原因.

  3. Effect of exogenous regulation of folate metabolism on L-serine accumulation of Corynebacterium glutamicum SYPS-062%外源调控叶酸代谢对谷氨酸棒杆菌SYPS-062积累L-丝氨酸的影响

    Institute of Scientific and Technical Information of China (English)

    徐国强; 袁圣男; 任建洪; 张晓梅; 许正宏

    2014-01-01

    通过外源添加叶酸合成前体对氨基苯甲酸、叶酸途径抑制剂氨甲喋呤扰动叶酸代谢,考察了叶酸对谷氨酸棒杆菌SYPS-062合成L-丝氨酸的影响.实验结果表明,添加对氨基苯甲酸(0~0.4 μg/mL)能够促进菌体的生长(最大值为7.8±0.24 g/L),但可显著降低其产酸能力;而添加氨甲喋呤(0~100 μg/mL)抑制了谷氨酸棒杆菌SYPS-062的生长,但可提高其单位菌体产酸的能力;代谢通量分析表明,作为调控点的叶酸代谢在SYPS-062生长及积累L-丝氨酸中占主导地位.

  4. Coryneform bacteria associated with canine otitis externa

    DEFF Research Database (Denmark)

    Aalbæk, Bent; Bemis, David A.; Schjærff, Mette;

    2010-01-01

    This study aims to investigate the occurrence of coryneform bacteria in canine otitis externa. A combined case series and case-control study was carried out to improve the current knowledge on frequency and clinical significance of coryneform bacteria in samples from canine otitis externa. A total...... of 16 cases of otitis externa with involvement of coryneform bacteria were recorded at two referral veterinary hospitals in Denmark and the US, respectively. Coryneform bacteria were identified by partial 16S rRNA gene sequencing. Corynebacterium auriscanis was the most common coryneform species (10...... cases). Small colony variants of this species were also observed. Other coryneform isolates were identified as Corynebacterium amycolatum (3 cases), Corynebacterium freneyi (2 cases) and an Arcanobacterium-like species (1 case). The coryneform bacteria were in all cases isolated together with other...

  5. LIQUID DYES'CHARACTERISTICS IN DYEING WASTE PAPER PULP AND THEIR APPLICATION

    Institute of Scientific and Technical Information of China (English)

    Xiaoping Wang; gang Chen; Aimin Tang; Hongwei Zhang

    2004-01-01

    In this paper, some liquid dyes were used to dye the waste paper pulp (OCC pulp and waste cement sack paper pulp), and their dyeing characteristics were analyzed, The liquid dyes include liquid basic yellow, liquid basic blue, liquid basic red, liquid basic orange, liquid basic brown and liquid direct black. We found that, each dye had its own dyeing characteristic while dyeing the waste paper pulp.Generally different types of liquid dyes were combined to dye the waste paper pulp, which the adding process must be noticed. We also observed that a black pigment could be applied together with said liquid dyes to dye or adjust the color of the bottom sheet for the fireproof board. We could also achieve the same dyeing result through different combinations of different dyes.

  6. LIQUID DYES'CHARACTERISTICS IN DYEING WASTE PAPER PULP AND THEIR APPLICATION

    Institute of Scientific and Technical Information of China (English)

    XiaopingWang; gangChen; AiminTang; HongweiZhang

    2004-01-01

    In this paper, some liquid dyes were used to dye the waste paper pulp (OCC pulp and waste cement sack paper pulp), and their dyeing characteristics were analyzed, The liquid dyes include liquid basic yellow, liquid basic blue, liquid basic red, liquid basic orange, liquid basic brown and liquid direct black. We found that, each dye had its own dyeing characteristic while dyeing the waste paper pulp. Generally different types of liquid dyes were combined to dye the waste paper pulp, which the adding process must be noticed. We also observed that a black pigment could be applied together withsaid liquid dyes to dye or adjust the color of the bottom sheet for the fireproof board. We could also achieve the same dyeing result through different combinations of different dyes.

  7. Gene transfer and transposition mutagenesis in Streptomyces roseosporus: mapping of insertions that influence daptomycin or pigment production.

    Science.gov (United States)

    McHenney, M A; Baltz, R H

    1996-09-01

    Streptomyces reseosporus, the producer of the cyclic lipopeptide antibiotic daptomycin, was shown to be a suitable host for molecular genetic manipulation. S. roseosporus does not appear to express significant restriction barriers based upon bacteriophage plaque formation studies. Plasmid DNA can be introduced into S. roseosporus by bacteriophage-FP43-mediated transduction and by conjugation from Escherichia coli. The streptomycete transposons Tn5096 and Tn5099, derived from IS493, transpose in S. roseosporus, and Tn5099-induced transposition mutants altered in the production of daptomycin, red pigment or black pigment were identified, and mapped to Dral and Asnl fragments. Three auxotrophic mutations (argB1, ade-1 and metB1) were identified among 100 individual Tn5096 insertions. Alignment and physical mapping of several Tn5099 insertions in Dral-E and Asnl-B fragments was facilitated by the presence of Dral and Asnl cleavage sites in Tn5099.

  8. Analytical Investigation Of Pigments, Ground Layer And Media Of Cartonnage Fragments From Greek Roman Period

    Science.gov (United States)

    Afifi, Hala. A. M.

    Some cartonnage fragments from Hawara, Fayoum Excavation were examined to identify pigments, media and grounds. It belonged to the Greek-Roman period. They were studied by X-ray diffraction (XRD), Energy dispersive X ray analysis (EDS) equipped with Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). These techniques were used to identify the composition and morphology of grounds, nature of pigments and media used in cartonnage fragments. The coarse ground layer was composed of calcite and traces of quartz. The fine ground layer used under the pigments directly was composed of calcite only. Carbon black was used as black pigment while lead oxide as red pigment, showing the influence of Roman and Greek pigments on Egyptian art in these later periods. Blue colorant was identified as cuprorivaite and yellow pigment was goethite. Animal glue was used in the four pigments as medium colored.

  9. Chromonychia Secondary to Chemotherapy

    Directory of Open Access Journals (Sweden)

    Marien Lopes

    2013-06-01

    Full Text Available Chemotherapy drugs can affect the skin and its appendages. Several clinical presentations can be observed, depending on the affected structure. The most common dermatological side effect is chromonychia. The main causative agents are: (1 cyclophosphamide, which can provoke a diffuse, black pigmentation, longitudinal striae and dark grey pigmentation located proximally on the nails; (2 doxorubicin, which promotes dark brown bands alternating with white striae and dark brown pigmentation in transverse bands, and (3 hydroxyurea, which produces a distal, diffuse, dark brown pigmentation. In the majority of cases, the effects are reversible after the suspension of the causative agent for a few months. We report a patient who developed chromonychia while undergoing treatment with cyclophosphamide, vincristine, doxorubicin, dexamethasone, methotrexate and cytarabine for acute lymphocytic leukemia.

  10. Spectrally selective paint coatings. Preparation and characterization

    Energy Technology Data Exchange (ETDEWEB)

    Crnjak Orel, Z.C.; Klanjsek Gunde, M. [National Institute of Chemistry, Hajdrihova 19, SI-1000 Ljubljana (Slovenia)

    2001-06-01

    Preparation and characterization of spectrally selective paint coating for photothermal solar energy conversion are discussed. The applied methods for preparation of paints with described measurements and calculations of black-pigmented coatings were reviewed. The article represents not only possible future applications but also past and current applications of spectrally selective paint coating which are used all over the world since the 1980s. Spectrally selective paint coatings based on combinations of two types of resins, various types of pigments and three types of silica, were prepared. The influence of pigment type and pigment volume concentration (PVC) was studied by applying the Kubelka-Munk (K-M) theory. The relation between the degrees of dispersion and distribution of pigment particles across the paint layer is discussed in terms of K-M coefficients.

  11. Phase and chemical composition analysis of pigments used in Cucuteni Neolithic painted ceramics

    Directory of Open Access Journals (Sweden)

    Bogdan Constantinescu

    2007-12-01

    Full Text Available Two analytical methods – 241Am-based X-Ray Fluorescence (XRF and Synchrotron Radiation X-ray Diffraction (SR-XRD – were used to investigate the elemental and mineralogical composition of pigments which decorate some Cucuteni Neolithic ceramic sherds. Local hematite and local calcite were the main components for red and white pigments, respectively. For black pigments, iron oxides (e.g. magnetite were used. They were often mixed with manganese oxides (e.g. jacobsite, which originated from Iacobeni manganese minerals deposits on the Bistrita River. Taking into account the results of the experiments, several conclusions regarding manufacturing procedures employed, and potential trade routes during the Neolithic were drawn.

  12. Develepment of Water-based Printing Inks for Corrugated Fibreboard%瓦楞纸用水性油墨的研制

    Institute of Scientific and Technical Information of China (English)

    崔久鑫

    2015-01-01

    以自制丙烯酸乳液为连结料,添加碳黑色浆,消泡剂,流平剂,增稠剂、成膜助剂制备了水性油墨,对水性油墨的细度、耐水性、光泽度等指标进行了测试,结果表明:水性油墨的各项指标符合工业标准。%The water-based Printing Inks were prepared by using acrylic acid emulsion as main binder , through adding carbon black pigments, antifoaming agents,leveling agent ,thicking agent and coalescent. The properties of the water soluble printing inks such as fineness ,water resistance ,and gloss were tested. The results indicated that all the properties are in accord with industry standards.

  13. [Hair as a study object in case of poisoning with heavy metal salts].

    Science.gov (United States)

    Pavlova, A Z; Bogomolov, D V; Larev, Z V; Amanmuradov, A Kh

    2012-01-01

    The objective of the present work was to analyse the results of morphological studies of hair taken from the children with suspected thallium poisoning. The findings obtained by isoelectrofocusing, spectroscopy, and scanning electron microscopy were compared with the relevant literature data. Original investigations included a comparative microstructural study along the hair length and cross section. We failed to observe formation of black-purple structures in the hair bulb and root region of the shaft usually associated with thallium poisoning. It is concluded that thallium poisoning can not be diagnosed based on the presence of a black pigment, knob-like swellings, and spindle-shaped bulbs since they are normal elements of healthy hair. More sensitive methods for the determination of trace elements and their combination with morphological investigations are needed for the definitive diagnosis of thallium poisoning.

  14. Nevus of ota with buccal mucosal pigmentation: a rare case.

    Science.gov (United States)

    Shetty, Shishir Ram; Subhas, Babu G; Rao, Kumuda Arvind; Castellino, Renita

    2011-01-01

    Nevus of Ota is a condition wherein the typical pattern of the bluish black pigmentation is noticed along with the cutaneous distribution of the trigeminal nerve. This condition is most prevalent in Japanese population but comparatively rare among Indians. We report a case of 23-year-old female presented with unilateral pigmented areas over the skin of forehead, malar area, ear and periorbital area. Blackish-blue pigmented areas were also noticed on the sclera. Brownish-black diffuse pigmented areas were also noticed on the buccal mucosa of the same side. The presence of pigmentation on the skin over pinna and oral pigmentation made our case a rare incidence. Oral pigmentations associated with nevus of Ota especially on the buccal mucosa have rarely been reported in the past.

  15. Nevus of Ota with buccal mucosal pigmentation: A rare case

    Directory of Open Access Journals (Sweden)

    Shishir Ram Shetty

    2011-01-01

    Full Text Available Nevus of Ota is a condition wherein the typical pattern of the bluish black pigmentation is noticed along with the cutaneous distribution of the trigeminal nerve. This condition is most prevalent in Japanese population but comparatively rare among Indians. We report a case of 23-year-old female presented with unilateral pigmented areas over the skin of forehead, malar area, ear and periorbital area. Blackish-blue pigmented areas were also noticed on the sclera. Brownish-black diffuse pigmented areas were also noticed on the buccal mucosa of the same side. The presence of pigmentation on the skin over pinna and oral pigmentation made our case a rare incidence. Oral pigmentations associated with nevus of Ota especially on the buccal mucosa have rarely been reported in the past.

  16. Distal-less homeobox genes of insects and spiders: genomic organization, function, regulation and evolution.

    Science.gov (United States)

    Chen, Bin; Piel, William H; Monteiro, Antónia

    2016-06-01

    The Distal-less (Dll) genes are homeodomain transcription factors that are present in most Metazoa and in representatives of all investigated arthropod groups. In Drosophila, the best studied insect, Dll plays an essential role in forming the proximodistal axis of the legs, antennae and analia, and in specifying antennal identity. The initiation of Dll expression in clusters of cells in mid-lateral regions of the Drosophila embryo represents the earliest genetic marker of limbs. Dll genes are involved in the development of the peripheral nervous system and sensitive organs, and they also function as master regulators of black pigmentation in some insect lineages. Here we analyze the complete genomes of six insects, the nematode Caenorhabditis elegans and Homo sapiens, as well as multiple Dll sequences available in databases in order to examine the structure and protein features of these genes. We also review the function, expression, regulation and evolution of arthropod Dll genes with emphasis on insects and spiders. PMID:26898323

  17. Porphyromonas gingivalis: An Overview of Periodontopathic Pathogen Below the Gum Line

    Directory of Open Access Journals (Sweden)

    Kah Yan eHow

    2016-02-01

    Full Text Available Periodontal disease represents a group of oral inflammatory infections initiated by oral pathogens which exist as a complex biofilms on the tooth surface and cause destruction to tooth supporting tissues. The severity of this disease ranges from mild and reversible inflammation of the gingiva (gingivitis to chronic destruction of connective tissues, the formation of periodontal pocket and ultimately result in loss of teeth. While human subgingival plaque harbors more than 500 bacterial species, considerable research has shown that Porphyromonas gingivalis, a Gram-negative anaerobic bacterium, is the major etiologic agent which contributes to chronic periodontitis. This black-pigmented bacterium produces a myriad of virulence factors that causes destruction to periodontal tissue either directly or indirectly by modulating the host inflammatory response. Here, this review provides an overview of P. gingivalis and how its virulence factors contribute to the pathogenesis with other microbiome consortium in oral cavity.

  18. Characterization of Sorolla's gouache pigments by means of spectroscopic techniques

    Science.gov (United States)

    Roldán, Clodoaldo; Juanes, David; Ferrazza, Livio; Carballo, Jorgelina

    2016-02-01

    This paper presents the characterization of the Joaquín Sorolla's gouache sketches for the oil on canvas series "Vision of Spain" commissioned by A. M. Huntington to decorate the library of the Hispanic Society of America in New York. The analyses were focused on the identification of the elemental composition of the gouache pigments by means of portable EDXRF spectrometry in a non-destructive mode. Additionally, SEM-EDX and FTIR analyses of a selected set of micro-samples were carried out to identify completely the pigments, the paint technique and the binding media. The obtained results have confirmed the identification of lead and zinc white, vermillion, earth pigments, ochre, zinc yellow, chrome yellow, ultramarine, Prussian blue, chromium based and copper-arsenic based green pigments, bone black and carbon based black pigments, and the use of gum arabic as binding media in the gouache pigments.

  19. Cutaneous Melanoma in Asians.

    Science.gov (United States)

    Kim, Sang Yub; Yun, Sook Jung

    2016-09-01

    Malignant melanoma is a rare disease in Asians but potentially the most aggressive form of skin cancer worldwide. It can occur in any melanocyte-containing anatomic site. Four main cutaneous melanoma subtypes are recognized: lentigo maligna melanoma, superficial spreading melanoma, acral lentiginous melanoma (ALM), and nodular melanoma. Generally, excessive exposure to ultraviolet (UV) radiation increases the risk of melanoma. The exception is ALM, which is the most common melanoma subtype in Asians and is not associated with UV radiation. ALM presents as dark brownish to black, irregular maculopatches, nodules, or ulcers on the palms, soles, and nails. The lesions may be misdiagnosed as more benign lesions, such as warts, ulcers, hematomas, foreign bodies, or fungal infections, especially in amelanotic acral melanomas where black pigments are absent. The aim of this brief review is to improve understanding and the rate of early detection thereby reducing mortality, especially regarding cutaneous melanoma in Asians. PMID:27689028

  20. Pseudomelanosis duodeni in a female adult with chronic renal failure

    Directory of Open Access Journals (Sweden)

    Wei-Chih Sun

    2014-12-01

    Full Text Available Pseudomelanosis duodeni is a rare endoscopic finding that manifests as dark speckled spots in the duodenum. It is considered a benign condition and is associated with certain diseases and the use of certain medications. This study reports a case of a 74-year-old woman, with end-stage renal disease under maintenance hemodialysis, hypertension under regular medical control, iron deficiency anemia under oral iron supplement, and progressive anemia with suspicious occult gastrointestinal bleeding. Upper gastrointestinal endoscopy revealed multiple tiny brownish-black pigmentation throughout the proximal second portion of the duodenum. The histopathological examination showed pigment-laden macrophages with positive iron stain and negative melanin stain in the lamina propria of the mucosal villi.

  1. Comparison of Riboflavin and Toluidine Blue O as Photosensitizers for Photoactivated Disinfection on Endodontic and Periodontal Pathogens In Vitro

    DEFF Research Database (Denmark)

    Nielsen, Henrik Krarup; Garcia, Javier; Væth, Michael;

    2015-01-01

    as a photosensitizer and blue LED light for activation, and compare it to photoactivated disinfection with the widely used combination of toluidine blue O and red light. Riboflavin is highly biocompatible and can be activated with LED lamps at hand in the dental office. To date, no reports are available......, Lactobacillus paracasei, Porphyromonas gingivalis, Prevotella intermedia and Propionibacterium acnes) were subjected to photoactivated disinfection with riboflavin/blue light and toluidine blue O/red light, and survival rates were determined by CFU counts. Within the limited irradiation time of one minute......, photoactivated disinfection with riboflavin/blue light only resulted in minor reductions in CFU counts, whereas full kills were achieved for all organisms when using toluidine blue O/red light. The black pigmented anaerobes P. gingivalis and P. intermedia were eradicated completely by riboflavin/blue light...

  2. Lasiodiplodia theobromae keratitis: a rare fungi from eastern India

    Directory of Open Access Journals (Sweden)

    Suman Saha

    2013-01-01

    Full Text Available We report here a case of a 32-year old female with a history of minor trauma presented with total corneal ulcer and hypopyon in the left eye. Microbiological examination of corneal scraping showed refractile hyphae with asepted branching filaments and black pigmented colonies in multiple solid agar medium. Identification of the organism was made from culture using D1/D2 region of Large Sub Unit (28S rDNA based molecular technique. Polymerase chain reaction amplified a band with a sequence that was 100% homologous with Lasiodiplodia theobromae. The organism was sensitive to amphotericin B and voriconazole, and demonstrated resistance to itraconazole and fluconazole. Therapeutic keratoplasty was performed followed by recurrence in graft controlled with topical voriconazole and intracameral amphotericin B. Graft failure was reported after three months.

  3. Studies on Romanian archaeological objects using atomic methods: the cases of Pietroasa hoard and Cucuteni ceramics

    International Nuclear Information System (INIS)

    can mention the following: - identification of black pigment composition from Cucuteni (Northern Moldova) and Ariusd (South-Eastern Transylvania) type pottery (VI - IV Millennia B. Chr.) as various combinations of goethite (alphaFeOOH), hausmannite (MnMn2O4) and bixbyte (Mn, Fe)2O3; - for high-temperatures (more than 600 deg C) fired pottery (the advanced Cucuteni ceramics types A and B) and psilomelane (MnO + MnO2 + H2O in variable proportions) for low temperatures (less than 400 deg C) fired pottery (the primitive pre-Cucuteni type C) - all these minerals originating from North Moldova mineral deposits of Iacobeni (neolithic trade routes put in evidence in this way - approx 500 km crossing Carpathian Mountains along Bistritza river); - no evidence of pyrolusite (MnO2) and manganite [MnO(OH)] - main components of Ukrainian Nikopol manganese deposit (used as black pigment source by contemporary Tripolye Neolithic culture) was found; - identification of magnetite (iron oxide Fe2 + Fe3 + O3) as main component for black pigments of Central Transylvania Petresti culture (4200 - 3500 B. Chr.); - identification of graphite as black pigment for Oltenia Starcevo-Cris culture - VI - V Millennia B. Chr.- ceramics (probably from Northern Bulgaria graphite deposits); - identification of organic origin (bones or wood) carbon-based pigments for few Cucuteni shards from North-Eastern Moldavia; - identification of white pigment composition as calcite (CaCO3) for Cucuteni culture and as calcium silicates mixed with illite (K, H2))Al2[(H2O, OH)2]AlSi3O10 for Petresti culture (Transylvania) and their minerals provenance areas; - identification of hematite (iron oxide Fe2O3) as main component for red pigments for all examined shards; - identification of all examined shards as having local provenance for the clay. (author)

  4. Studies on Romanian archaeological objects using nuclear methods: cases of Pietroasa hoard and Cucuteni ceramics

    International Nuclear Information System (INIS)

    mention: - identification of black pigment composition from Cucuteni (northern Moldova) and Ariusd (south-eastern Transylvania) type pottery (VI - IV Millennia B. Chr.) as various combinations of goethite (alphaFeOOH), hausmannite (MnMn2O4) and bixbyte (Mn, Fe)2O3 - for high-temperatures ( more than 600 deg C) fired pottery (the advanced Cucuteni ceramics types A and B) and psilomelane (MnO + MnO2 + H2O in various concentration ratios) for low temperatures (less than 400 deg C) fired pottery (the primitive pre-Cucuteni type C) - all these minerals originating from northern Moldova mineral deposits of Iacobeni (neolithic trade routes put in evidence in this way - approx 500 km crossing Carpathian Mountains along Bistrita river); - no evidence of pyrolusite (MnO2) and manganite [MnO(OH)] - main components of Ukrainian Nikopol manganese deposit (used as black pigment source by contemporary Tripolye Neolithic culture) was found; - identification of magnetite (iron oxide Fe2+Fe3+O3) as main component for black pigments of Central Transylvania Petresti culture (4200 - 3500 B. Chr.); - identification of graphite as black pigment for Oltenia Starcevo-Cris culture - VI - V Millennia B. Chr.- ceramics (probably from northern Bulgaria graphite deposits); - identification of organic origin (bones or wood) carbon-based pigments for few Cucuteni shards from north-eastern Moldova; - identification of white pigment composition as calcite (CaCO3) for Cucuteni culture and as calcium silicates mixed with illite [(K, H2))Al2[(H2O, OH)2]AlSi3O10] for Petresti culture (Transylvania) and their minerals provenance areas; - identification of hematite (iron oxide Fe2O3) as main component for red pigments for all examined shards; - identification of all examined shards as having local provenance for the clay. (authors)

  5. The effect of blue light on periodontal biofilm growth in vitro.

    Science.gov (United States)

    Fontana, Carla R; Song, Xiaoqing; Polymeri, Angeliki; Goodson, J Max; Wang, Xiaoshan; Soukos, Nikolaos S

    2015-11-01

    We have previously shown that blue light eliminates the black-pigmented oral bacteria Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, and Prevotella melaninogenica. In the present study, the in vitro photosensitivity of the above black-pigmented microorganisms and four Fusobacteria species (Fusobacterium nucleatum ss. nucleatum, F. nucleatum ss. vincentii, F. nucleatum ss. polymorphum, Fusobacterium periodonticum) was investigated in pure cultures and human dental plaque suspensions. We also tested the hypothesis that phototargeting the above eight key periodontopathogens in plaque-derived biofilms in vitro would control growth within the dental biofilm environment. Cultures of the eight bacteria were exposed to blue light at 455 nm with power density of 80 mW/cm2 and energy fluence of 4.8 J/cm2. High-performance liquid chromatography (HPLC) analysis of bacteria was performed to demonstrate the presence and amounts of porphyrin molecules within microorganisms. Suspensions of human dental plaque bacteria were also exposed once to blue light at 455 nm with power density of 50 mW/cm2 and energy fluence of 12 J/cm2. Microbial biofilms developed from the same plaque were exposed to 455 nm blue light at 50 mW/cm2 once daily for 4 min (12 J/cm2) over a period of 3 days (4 exposures) in order to investigate the cumulative action of phototherapy on the eight photosensitive pathogens as well as on biofilm growth. Bacterial growth was evaluated using the colony-forming unit (CFU) assay. The selective phototargeting of pathogens was studied using whole genomic probes in the checkerboard DNA-DNA format. In cultures, all eight species showed significant growth reduction (p biofilms, respectively, (p biofilms. The cumulative blue light treatment suppressed biofilm growth in vitro. This may introduce a new avenue of prophylactic treatment for periodontal diseases. PMID:25759232

  6. The effect of blue light on periodontal biofilm growth in vitro.

    Science.gov (United States)

    Fontana, Carla R; Song, Xiaoqing; Polymeri, Angeliki; Goodson, J Max; Wang, Xiaoshan; Soukos, Nikolaos S

    2015-11-01

    We have previously shown that blue light eliminates the black-pigmented oral bacteria Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, and Prevotella melaninogenica. In the present study, the in vitro photosensitivity of the above black-pigmented microorganisms and four Fusobacteria species (Fusobacterium nucleatum ss. nucleatum, F. nucleatum ss. vincentii, F. nucleatum ss. polymorphum, Fusobacterium periodonticum) was investigated in pure cultures and human dental plaque suspensions. We also tested the hypothesis that phototargeting the above eight key periodontopathogens in plaque-derived biofilms in vitro would control growth within the dental biofilm environment. Cultures of the eight bacteria were exposed to blue light at 455 nm with power density of 80 mW/cm2 and energy fluence of 4.8 J/cm2. High-performance liquid chromatography (HPLC) analysis of bacteria was performed to demonstrate the presence and amounts of porphyrin molecules within microorganisms. Suspensions of human dental plaque bacteria were also exposed once to blue light at 455 nm with power density of 50 mW/cm2 and energy fluence of 12 J/cm2. Microbial biofilms developed from the same plaque were exposed to 455 nm blue light at 50 mW/cm2 once daily for 4 min (12 J/cm2) over a period of 3 days (4 exposures) in order to investigate the cumulative action of phototherapy on the eight photosensitive pathogens as well as on biofilm growth. Bacterial growth was evaluated using the colony-forming unit (CFU) assay. The selective phototargeting of pathogens was studied using whole genomic probes in the checkerboard DNA-DNA format. In cultures, all eight species showed significant growth reduction (p biofilms, respectively, (p biofilms. The cumulative blue light treatment suppressed biofilm growth in vitro. This may introduce a new avenue of prophylactic treatment for periodontal diseases.

  7. Synthesis and characterization of black, red and yellow nanoparticles pigments from the iron sand

    Energy Technology Data Exchange (ETDEWEB)

    Mufti, Nandang, E-mail: nandangmufti@gmail.com; Atma, T., E-mail: nandangmufti@gmail.com; Fuad, A., E-mail: nandangmufti@gmail.com [Department of Physics, University of Malang, Jl Semarang-65145, Malang (Indonesia); Sutadji, E. [Department of Mechanical Engineering, University of Malang, Jl Semarang-65145, Malang (Indonesia)

    2014-09-25

    The aim of this research is to synthesize nanoparticles of black pigment of Magnetite (Fe{sub 3}O{sub 4}), red pigment of hematite (α-Fe{sub 2}O{sub 3}), and yellow pigment of ghoetite (α-FeOOH) from the iron sand. The black pigment of Fe{sub 3}O{sub 4} and the yellow pigment α-FeOOH nanoparticles were synthesized by coprecipitation method with variation of pH. Whereas, the red pigment Fe{sub 2}O{sub 3} was synthesized by sintering Fe{sub 3}O{sub 4} nanoparticles at temperature between 400 °C and 700 7°C for 1 hour. All the pigments has been characterized using X-ray diffraction and SEM. The XRD results shown that the particle size of the black pigmen Fe{sub 3}O{sub 4}, red pigment Fe{sub 3}O{sub 4} and yellow pigment α-FeOOH are around 12, 32, and 30 nm respectively. The particle size of Fe{sub 2}O{sub 3} nanoparticles increase by increasing sintering temperature from 32 nm at 400 °C to 39 nm at 700 °C. For yellow pigment of α-FeOOH, the particle size increase by increasing pH from 30,54 nm at pH 4 to 48,60 nm at pH 7. The SEM results shown that the morphologies of black, yellow and red pigments are aglomarated.

  8. PENGEMBANGAN INOKULUM UNTUK MENINGKATKAN MUTU GIZI TEMPE

    Directory of Open Access Journals (Sweden)

    Mien Karmini

    2012-11-01

    Full Text Available Inokulum tempe yang lazim digunakan di Indonesia mengandung Rhizopus Sp. Penelitian terdahulu menemukan dua jenis bakteri penghasil vitamin B12, yaitu Citrobacter freundil dan Klebsiella pneumoniae, dan dua jenis bakteri penghasil Faktor-2, yaitu Corynebacterium Sp dan Micrococcus luteus dari air rendaman kedelai. Dalam penelitian ini dibuat inokulum tempe yang merupakan campuran kapang Rhizopus oligosporus dan masing-masing bakteri Klebsiella pneumoniae atau Citrobacter freundii mampu meningkatkan kadar vitamin B12 di dalam tempe lebih dari 100%. Inokulum campuran Rhizopus oligosporus dan Corynebacterium Sp atau Micrococcus luteus tidak menunjukkan adanya kandungan antioksidan Faktor-2 di dalam tempe.

  9. [Talc--a standard drug in the treatment of malignant pleural effusion].

    Science.gov (United States)

    Bednár, M; Bezdícek, P

    1999-01-25

    Currently, talc is the most effective and widely-used agent for chemical pleurodesis in the treatment of both malignant pleural effusion and pneumothorax. Its popularity has been growing due to the low incidence of side effects, low cost and higher success rate in comparison with other agents (tetracyclines, bleomycin, Corynebacterium parvum). The guidelines for talc pleurodesis are summarized and the history of the Corynebacterium parvum as an agent for chemical pleurodesis is mentioned. Its production in the Czech Republic has been halted due to the increased interest in the talc as a chemical sclerosant. PMID:10376391

  10. GenBank blastx search result: AK287424 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287424 J043006K04 AF414084.1 AF414084 Corynebacterium crenatum feedback-resistant... aspartokinase LysC alpha subunit and feedback-resistant aspartokinase LysC beta subunit genes, complete cds. BCT 5e-24 0 ...

  11. GenBank blastx search result: AK061941 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061941 001-042-D10 AF414084.1 Corynebacterium crenatum feedback-resistant asparto...kinase LysC alpha subunit and feedback-resistant aspartokinase LysC beta subunit genes, complete cds.|BCT BCT 2e-34 +2 ...

  12. Experimental otitis media in gerbils and chinchillas with Streptococcus pneumoniae, Haemophilus influenzae, and other aerobic and anaerobic bacteria.

    OpenAIRE

    Fulghum, R S; Brinn, J E; Smith, A M; Daniel, H J; Loesche, P J

    1982-01-01

    To ascertain the usefulness of Mongolian gerbils as an inbred model for otitis media, 52 Mongolian gerbils (Meriones unguiculatus, strain MONT/Tum) were compared with 26 chinchillas (Chinchilla laniger) for susceptibility to Streptococcus pneumoniae type 3. Haemophilus influenzae type b, and a polymicrobic culture including anaerobes (Streptococcus intermedius, Propionibacterium acnes, Staphylococcus epidermidis, and Corynebacterium sp.). Organisms were inoculated percutaneously into the supe...

  13. [Method and procedures in bacteriological study of necrotic teeth].

    Science.gov (United States)

    Rodríguez-Ponce, A; López Campos, A; López Paz, J; Pazos Sierra, R

    1991-01-01

    Research was conducted of 160 radicular canals with necrotic pulp. Results of different bacteriological analyses are presented. Culture analyses in aerobic and anaerobic media, resulted in the isolation of Staphylococcus Epidermidis, Streptococcus Viridans and Corynebacterium sp in the group studied, as the most frequent bacteria. There was no evidence of a specific germ linked with the pulp necrosis. PMID:1659855

  14. Antibacterial and antifungal activities of different parts of Tribulus terrestris L. growing in Iraq

    OpenAIRE

    AL-BAYATI, Firas A.; Al-Mola, Hassan F.

    2008-01-01

    Antimicrobial activity of organic and aqueous extracts from fruits, leaves and roots of Tribulus terrestris L., an Iraqi medicinal plant used as urinary anti-infective in folk medicine, was examined against 11 species of pathogenic and non-pathogenic microorganisms: Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Corynebacterium diphtheriae, Escherichia coli, Proteus vulgaris, Serratia marcescens, Salmonella typhimurium, Klebsiella pneumoniae, Pseudomonas aeruginosa and Candida alb...

  15. Disease: H01051 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available i A, Azevedo V, Schneider MP, Silva A, Santos CS, Santos LS, Sabbadini P, Dias AA, Hirata R Jr, Mattos-Guaraldi AL, Tauch A Comparati...ve analysis of two complete Corynebacterium ulcerans genomes and detection of candidate virulence factors. BMC Genomics 12:383 (2011) ...

  16. Activity of autoinducer two (AI-2) in bacteria isolated from surface ripened cheeses

    DEFF Research Database (Denmark)

    Gori, Klaus; Jespersen, Lene

    ). Corynebacterium casei, Microbacterium barkeri, Microbacterium gubbeenense and S. equorum subsp. linens (all isolated from the smear of surface ripened cheeses) using the AI-2 bioluminescence assay. This indicates that AI-2 signaling could take place between bacteria found in the smear of surface ripened cheeses....

  17. Activity of autoinducer two (AI-2) in bacteria isolated from surface ripened cheeses

    DEFF Research Database (Denmark)

    Gori, Klaus; Jespersen, Lene

    2007-01-01

    ). Corynebacterium casei, Microbacterium barkeri, Microbacterium gubbeenense and S. equorum subsp. linens (all isolated from the smear of surface ripened cheeses) using the AI-2 bioluminescence assay. This indicates that AI-2 signaling could take place between bacteria found in the smear of surface ripened cheeses....

  18. Polyamine Distribution Patterns in Coryneform Bacteria and Related Gram-Positive Eubacteria

    OpenAIRE

    Hamana, Koei

    1996-01-01

    Polyamines of the nine genera of coryneform and related Gram-positive eubacteria were analyzed by HPLC. Authentic species of Microbacterium, Aureobacterium, Cellulomonas and Corynebacterium were devoid of polyamines. Arthrobacter species were divided into polyamine-absent, putrescine, cadaverine and putrescine-cadaverine types. Clavibacter contained putrescine and cadaverine. Spermidine was detected in some species of Brevibacterium, Exiguobacterium and Curtobacterium and diaminopropane in so...

  19. [Method and procedures in bacteriological study of necrotic teeth].

    Science.gov (United States)

    Rodríguez-Ponce, A; López Campos, A; López Paz, J; Pazos Sierra, R

    1991-01-01

    Research was conducted of 160 radicular canals with necrotic pulp. Results of different bacteriological analyses are presented. Culture analyses in aerobic and anaerobic media, resulted in the isolation of Staphylococcus Epidermidis, Streptococcus Viridans and Corynebacterium sp in the group studied, as the most frequent bacteria. There was no evidence of a specific germ linked with the pulp necrosis.

  20. Sexually transmitted diphtheria.

    Science.gov (United States)

    Berger, Anja; Lensing, Carmen; Konrad, Regina; Huber, Ingrid; Hogardt, Michael; Sing, Andreas

    2013-03-01

    Diphtheria is caused by diphtheria toxin-producing Corynebacterium species. While classical respiratory diphtheria is transmitted by droplets, cutaneous diphtheria often results from minor trauma. This report concerns the first case of sexually transmitted diphtheria in a patient with non-gonococcal urethritis after orogenital contact. PMID:22628666

  1. Biochemical aspects of the immunomodular action in irradiated survival mice with 60C gama irradiation

    International Nuclear Information System (INIS)

    The radioprotective action of Calmetti-Guerin bacillus (BCG), Corynebacterium parvum, Escherichia coli Lipopolysccharides (LPS) and peptone proteose was evaluated. A single injection of the macrophage activiting agents prior to 60Co whole-body irradiation increased the survival rate of mice in the lethal dose range. (L.M.J.)

  2. A serological investigation of caseous lymphadenitis in four flocks of sheep

    NARCIS (Netherlands)

    Malone, F.E.; Fee, S.A.; Kamp, E.M.; King, D.C.; Baird, G.J.; O'Reilly, K.M.; Murdock, F.E.A.

    2006-01-01

    A double antibody sandwich ELISA developed by ID-DLO, Lelystad to detect Corynebacterium pseudotuberculosis infection was used on 329 sheep from four pedigree Suffolk flocks in which clinical cases of caseous lymphadenitis (CLA) had occurred. At subsequent necropsy, typical CLA lesions were seen in

  3. In vitro anti-tumour activity of tumour necrosis serum

    NARCIS (Netherlands)

    Bloksma, N.; Schetters, Th.P.; Figdor, C.; Dijk, H. van; Willers, J.M.

    1980-01-01

    A method measuring 3H-thymidine incorporation in Meth A sarcoma cells was used to quantify in vitro anti-tumour activity of tumour necrosis serum and compared with a method using cell viability as a parameter. Tumour necrosis serum obtained from mice pretreated with Corynebacterium parvum and elicit

  4. [Current treatment strategy in malignant pleural effusion].

    Science.gov (United States)

    Türler, A; Walter, M; Schmitz-Rixen, T

    1996-01-01

    Malignant pleural effusions are a grave consequence of advanced cancer disease. The successful suppression of pleural fluid reaccumulation can make a major contribution to the management and palliative care of patients with disseminated cancer. Many treatment concepts have been reported in the literature. The recommended therapy in malignant pleural effusions consists of intrapleural instillation of a sclerotic agent to produce pleurodesis. Different substances have been used, including tetracyclines, cytostatic agents, fibrin, talc, Corynebacterium parvum, cytokines and others. We reviewed the most frequently used techniques of pleurodesis in order to define the most effective treatment concept. In 15 prospective randomized trials the success rates varied from 13% with bleomycin to 100% with talc or Corynebacterium parvum. Talc was superior to other agents in 6 of 6, Corynebacterium parvum in 3 of 4 and bleomycin or tetracycline only in 3 of 8 studies. Adverse effects were frequently observed with cytostatic agents, but were very rare in the case of talc or fibrin instillation. Comparing the recently published data pleurodesis with talc appears to be the most effective treatment strategy, followed by Corynebacterium parvum, bleomycin and tetracycline. PMID:8686317

  5. Domain Modeling: NP_003264.2 [SAHG[Archive

    Lifescience Database Archive (English)

    Full Text Available NP_003264.2 chr11 CRYSTAL STRUCTURE OF THE SODIUM-COUPLED GLYCINE BETAINE SYMPORTER... BETP FROM CORYNEBACTERIUM GLUTAMICUM WITH BOUND SUBSTRATE p2wita_ chr11/NP_003264.2/NP_003264.2_holo_12-404.pdb swppa 110A,111L,113V,115L,130L,134A,135F,258M BET 0 ...

  6. Occurrence of Listeria spp. in farmed salmon and during subsequent slaughter: Comparison of listertest TM lift and the USDA method

    DEFF Research Database (Denmark)

    Embarek, Peter Karim Ben; Hansen, Lisbeth Truelstrup; Enger, O.;

    1997-01-01

    Salmon and environmental samples from a fish farm and a salmon slaughterhouse were analysed for Listeria spp. using the USDA method and the rapid quantitative Listertest(TM). Listeria spp. were not detected in any of 119 samples analysed. However, Corynebacterium spp. gave disturbing false...

  7. 21 CFR 573.500 - Condensed, extracted glutamic acid fermentation product.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Condensed, extracted glutamic acid fermentation... fermentation product. Condensed, extracted glutamic acid fermentation product may be safely used in animal feed... the extraction of glutamic acid, combined with the cells of Corynebacterium lilium used to produce...

  8. Cutaneous ulcers in a returning traveller: a rare case of imported diphtheria in the UK.

    Science.gov (United States)

    Nelson, T G; Mitchell, C D; Sega-Hall, G M; Porter, R J

    2016-01-01

    We describe a case of cutaneous diphtheria in the UK, presenting as lower leg ulcers in a returning traveller, and discuss the epidemiology, significance and public health implications of this disease and the therapeutic options available. A 65-year-old woman presented with a 6-week history of multiple ulcers appearing on her legs following a holiday in Kenya. Culture of biopsy tissue grew Corynebacterium diphtheriae. A cascade of therapeutic and public health interventions followed, many of which were terminated once the isolate was confirmed as nontoxigenic. Cutaneous diphtheria is a rare, notifiable disease in the UK, but is common in tropical countries, and is most often seen in the West as a traveller's disease. Corynebacteria are common skin commensals, and without appropriate clinical details, laboratories may not recognize C. diphtheriae/Corynebacterium ulcerans. This is likely to have led to under-reporting and under-recognition of the condition. PMID:26455435

  9. 几株城市污水优势降解细菌的初步鉴定%Preliminary Identification of Several Predominant Degradation Strains for Municipal Sewage Treatment

    Institute of Scientific and Technical Information of China (English)

    刘正学; 刘云峰; 李德舜; 张英

    2007-01-01

    通过革兰氏等染色、个体及菌苔形态特征观察、生理生化反应等研究方法,对6株分离自城市污水的优势高效降解细菌,即9、X-18、M-28、M-6、X-39和X-58进行了分类鉴定.结果表明:它们分别隶属于色杆菌属(Chromobacterium sp.)、假单细胞菌属(Pseudomonas sp.)、短杆菌属(Brevibacterium sp.)、节杆菌属(Arthobacter sp.)、棒杆菌属(Corynebacterium sp.)、棒杆菌属(Corynebacterium sp.).

  10. [Chest drainage combined with intracavitary therapy in neoplastic pleural effusion: comparison of three different protocols].

    Science.gov (United States)

    D'Angelo, P; Sacco, R; Romessis, M; Mucilli, F

    1993-07-01

    Twenty-eight patients with malignant pleural effusion observed in a two year period were treated with intrapleural instillation of different substances: Tetracycline, Corynebacterium parvum and Beta-Interferon. Different results were observed: complete responsiveness (no recurrence of pleural effusion within three months); partial responsiveness (recurrence of moderate pleural effusion within one month after drainage removal); insufficient responsiveness (recurrence of massive effusion within one month). Among patients treated with Tetracycline seven complete, five partial and one insufficient responses were observed. Instillation of Corynebacterium parvum allowed two complete, two partial and three insufficient responses. Finally, in the group treated with Beta-Interferon complete responsiveness was obtained in just one patient, partial responsiveness in three, while the treatment was insufficient in the last two. These results suggest pleural drainage is the best treatment in patients with malignant pleural effusion, however, the association of Tetracycline instillation allows better results. PMID:8398623

  11. Preliminary study of the flora in the lower genital tracts of sexually active adolescent females in relation to symptoms and inflammatory response.

    Science.gov (United States)

    Russo, J F; Ronkin, S; Furness, G

    1981-03-01

    The lower genital tracts of 20 sexually active adolescent females were examined for the presence of Mycoplasma hominis, Ureaplasma urealyticum, Corynebacterium genitalium, and Corynebacterium pseudogenitalium. Fifty percent of the asymptomatic adolescent females and 50% of the symptomatic adolescent females were colonized with M. hominis, which is higher than the percentage reported in adult females. None of the asymptomatic adolescent females and only 10% of the symptomatic adolescent females were colonized with U. urealyticum, which is much lower than the percentage reported in adults. None of the study patients was colonized with C. genitalium, but 25% were colonized with C. pseudogenitalium. No relationship was found between the presence of M. hominis in the lower genital tract and a clinically identifiable vaginal discharge or inflammatory changes in exfoliated cervical and vaginal epithelial cells. The presence of M.hominis in the lower genital tract does not appear to be influenced by use of oral contraceptives or antecedent pregnancy. PMID:7333925

  12. Iris rubeosis and hyphema caused by chemical injury due to household detergent

    Directory of Open Access Journals (Sweden)

    Suto C

    2012-11-01

    Full Text Available Chikako Suto,1,2 Tetsuya Ishizuka,1 Hiroshi Toshida31Department of Ophthalmology, Saiseikai Kurihashi Hospital, Kuki, Saitama, 2Department of Ophthalmology, Tokyo Women's Medical University, Shinjuku, Tokyo, 3Department of Ophthalmology, Juntendo University Shizuoka Hospital, Izunokuni, Shizuoka, JapanAbstract: We report an unusual case of iris rubeosis and hyphema caused by chemical injury due to household detergent. A 74-year-old man with a 15-year history of diabetes mellitus was refilling a container with household detergent at home. He splashed the detergent in his eyes. Slit-lamp examination revealed extensive epithelial damage to the left eye, leading to a persistent corneal epithelial defect. We used a bandage soft contact lens with levofloxacin eye drops as concomitant therapy in order to promote healing. However, a strain of fluoroquinolone-resistant Corynebacterium colonized the eye, so that the corneal ulcer eventually became severe. Use of the bandage soft contact lens was discontinued. His antimicrobial agent was changed to cefmenoxime, a drug to which fluoroquinolone-resistant Corynebacterium is sensitive, and topical instillation of autologous serum subsequently promoted improvement of the ulcer. On day 38 after injury, iris rubeosis led to hyphema and ghost cell glaucoma. With improvement of his corneal epithelial defect, the iris rubeosis and hyphema regressed and his visual acuity improved to 20/25 on the left eye. To the best of our knowledge, this is the first report of a case resulting in severe complications due to chemical injury by a neutral detergent. Ophthalmologists should be aware that corneal epithelial damage may become prolonged in elderly patients with diabetes, and unexpectedly severe when wearing bandage soft contact lens, with infection of Corynebacterium resistant to fluoroquinolones, even if the chemical agent is a neutral detergent.Keywords: chemical injury, household detergent, persistent corneal

  13. Antimicrobial Activity of Six Pomegranate (Punica granatum L.) Varieties and Their Relation to Some of Their Pomological and Phytonutrient Characteristics

    OpenAIRE

    Nurcan Erbil; Kenan S. Dayisoylu; Mustafa Ozgen; Ahmet D. Duman; Coskun Durgac

    2009-01-01

    Arils from six pomegranate (Punica granatum L.) varieties grown in the Mediterranean region of Turkey were tested for their antimicrobial properties by the agar diffusion and minimum inhibitory concentration (MIC) methods against seven bacteria: (Bacillus megaterium DSM 32, Pseudomonas aeruginosa DSM 9027, Staphylococcus aureus Cowan 1, Corynebacterium xerosis UC 9165, Escherichia coli DM, Enterococcus faecalis A10, Micrococcus luteus LA 2971), and threefungi (Kluvyeromyces marxianus A230, Rh...

  14. The dynamics of bacteria population on the skin, throat, and gastrointestinal tract of HIV-seropositive patients

    Directory of Open Access Journals (Sweden)

    Kwashie Ajibade Ako-Nai

    2015-01-01

    Full Text Available Background: The study determined bacteria population on the skin, throat, and gastrointestinal tract of human immunodeficiency virus (HIV-seropositive patients and HIV seronegative controls at the baseline, 3 months, and 6 months, respectively, at Obafemi Awolowo University Teaching Hospitals Complex (OAUTHC, Ilé-Ifè, Osun State, Nigeria and State Specialist Hospital, Akure, Ondo State, Nigeria between May and November, 2012. Materials and Methods: Seventy HIV-seropositive subjects and 51 HIV seronegative controls who attended the HIV clinics were recruited. Skin, throat, and rectal swabs were obtained from the participants using sterile cotton-tipped applicators introduced into thioglycollate broth and incubated at 37°C overnight. When growth was noticed, the broth culture was streaked on different bacteriologic media and the isolates were characterized by the standard methods and disc diffusion for antibiotic sensitivity. Results: The number of isolates cultured from the HIV-seropositive subjects was 934, with the distribution being 397, 326, and 211 at the baseline, 3 months, and 6 months, respectively. The distribution of 1,138 isolates cultured from 51 HIV-seronegative controls was 433, 354, and 351 at the baseline, 3 months, and 6 months, respectively. At the baseline among HIV-seropositive patients, the predominant isolates were Arcanobacterium haemolyticum, Pseudomonas aeruginosa (P. aeruginosa, and Bacillus cereus (B. cereus. However, Corynebacterium haemolyticum, Enterococcus faecalis, and Escherichia coli (E. coli were predominant at 3 months while at 6 months, Corynebacterium haemolyticum and Corynebacterium diphtheriae had the highest frequency followed by Pseudomonas fluorescens (P. fluorescens. In the controls, Corynebacterium diphtheriae, Listeria monocytogenes, and Staphylococcus xylosus (S. xylosus predominated at the baseline and at 3 months while at 6 months, B. cereus, S. xylosus, and Staphylococcus aureus (S. aureus were

  15. Identification of plasmid partition function in coryneform bacteria.

    OpenAIRE

    Kurusu, Y; Satoh, Y.; Inui, M.; Kohama, K; Kobayashi, M.; Terasawa, M.; Yukawa, H

    1991-01-01

    We have identified and characterized a partition function that is required for stable maintenance of plasmids in the coryneform bacteria Brevibacterium flavum MJ233 and Corynebacterium glutamicum ATCC 31831. This function is localized to a HindIII-NspV fragment (673 bp) adjacent to the replication region of the plasmid, named pBY503, from Brevibacterium stationis IFO 12144. The function was independent of copy number control and was not associated directly with plasmid replication functions. ...

  16. Transcriptional analysis and regulatory signals of the hom-thrB cluster of Brevibacterium lactofermentum.

    OpenAIRE

    Mateos, L M; Pisabarro, A; Pátek, M; Malumbres, M; Guerrero, C.; Eikmanns, B J; Sahm, H; Martín, J F

    1994-01-01

    Two genes, hom (encoding homoserine dehydrogenase) and thrB (encoding homoserine kinase), of the threonine biosynthetic pathway are clustered in the chromosome of Brevibacterium lactofermentum in the order 5' hom-thrB 3', separated by only 10 bp. The Brevibacterium thrB gene is expressed in Escherichia coli, in Brevibacterium lactofermentum, and in Corynebacterium glutamicum and complements auxotrophs of all three organisms deficient in homoserine kinase, whereas the Brevibacterium hom gene d...

  17. Isolation and Preliminary Characterization of Twenty Bacteriophages Infecting Either Brevibacterium or Arthrobacter Strains

    OpenAIRE

    Trautwetter, Annie; Blanco, Carlos

    1988-01-01

    Thirty-seven bacteriophages plaquing on Corynebacterium, Brevibacterium, or Arthrobacter strains were isolated from soil or vegetation samples. Restriction analysis of phage DNA indicated that 20 phages were unique; one of them produced entirely turbid plaques on Brevibacterium ketoglutamicum and was characterized as temperate. All these phages were assigned to group B of the classification of Bradley (Bacteriol. Rev. 31:230-314, 1967) and had relatively narrow host ranges.

  18. МУТАНТНЫЕ ШТАММЫ МИКРООРГАНИЗМОВ ПРОДУЦЕНТОВ ЛИЗИНА И ТРЕОНИНА

    OpenAIRE

    Андрияш, Г.; Заболотная, Г.; Шульга, С.

    2014-01-01

    Strains-producers of essential amino acids of aspartate family such as Corynebacterium glutamicum, Brevibacterium flavum, Brevibacterium sp. 90, Brevibacterium sp. 90H, Brevibacterium sp. E531 from «Collections strains and lines of plants for food and agricultural biotechnology» of «Institute of Food Biotechnology and Genomics of the National Academy of Sciences of Ukraine» for biosynthetic activity for lysine and threonine were investigated. Active strains-producers of amino acids were obtai...

  19. Non-toxigenic penicillin-resistant cutaneous C. diphtheriae infection: a case report and review of the literature.

    Science.gov (United States)

    FitzGerald, Rosemarie Philippa; Rosser, Andrew J; Perera, Dona Nelun

    2015-01-01

    Here, we report a case of non-toxigenic Corynebacterium diphtheriae in a previously healthy 14-year-old girl that was acquired in Ethiopia and presented locally. This is the first clinical case of penicillin-resistant C. diphtheriae in the UK. This is significant finding because penicillin is the recommended first-line agent for the prophylaxis against and treatment of C. diphtheriae in patients who are not allergic to penicillin. PMID:25027172

  20. 发酵液中L-谷氨酰胺含量的测定%The Quantitative Determinations of L-Glutamine in Fermanted Liquor

    Institute of Scientific and Technical Information of China (English)

    汪世华; 白文钊; 康旭; 吴思方

    2001-01-01

    对谷氨酸产生菌S9114摇瓶发酵72 h后,用纸层析法和荧光法定量测定发酵液中L-谷氨酰胺含量.%Corynebacterium Pekinense S9114,a Glutamine acid producer,is selected for this study.After 72 hours shaking cultivation,we have reported the quantitative detetrminations of L-Glutamine by paper chomatography and fluorometry.

  1. 短小棒状杆菌治疗恶性胸水的临床观察

    Institute of Scientific and Technical Information of China (English)

    牟旭日; 姜文升; 李瑞杰

    2004-01-01

    恶性胸水是晚期恶性肿瘤,尤其是肺癌和乳腺癌的常见并发症,自1997年5月—2002年8月,我院对96例癌性胸水患者应用短小棒状杆菌(Corynebacterium Parvum Vaccine,以下简称CPV)治疗,取得显著疗效。

  2. 胸腔内注入榄香烯或短小棒状杆菌菌苗治疗恶性胸腔积液

    Institute of Scientific and Technical Information of China (English)

    白冲; 刘忠令; 王昌惠; 韩一平

    1999-01-01

    @@ 肺癌或其他癌肿转移至胸膜易导致恶性胸腔积液.我院于1994~1998年胸腔内分别注射榄香烯和短小棒状杆菌菌苗(Corynebacterium parvum,CP)治疗恶性胸腔积液82例,现报告如下.

  3. Protective effect of Lactobacillus casei on Pseudomonas aeruginosa infection in mice.

    OpenAIRE

    Miake, S; Yokokura, T; Yoshikai, Y; Mutai, M; Nomoto, K.

    1985-01-01

    The protective effect of heat-killed Lactobacillus casei YIT9018 (LC 9018) against Pseudomonas aeruginosa infection in mice was compared with that of Corynebacterium parvum. Survival of mice after intraperitoneal (i.p.) infection with P. aeruginosa was augmented in mice that had been pretreated i.p. with LC 9018 5 days previously. Similar treatment of mice with C. parvum, however, was not effective at all. Moreover, mice became more susceptible to infection with P. aeruginosa after such treat...

  4. A trial of nonspecific immunotherapy using systemic C. parvum in treated patients with Dukes B and C colorectal cancer.

    OpenAIRE

    Souter, R. G.; Gill, P. G.; Morris, P.J.

    1982-01-01

    In view of the relatively poor prognosis for patients after surgery for locally invasive colorectal cancer a trial of repeated systemic infusions of Corynebacterium parvum (CP) has been carried out. It is in this group of patients, with a high risk of recurrence from small residues of cancer left by the surgeon, that immunotherapy should have its optimum chance of success. A total of 92 patients were included in a randomized controlled study. The two groups were comparable in terms of tumour ...

  5. Mechanisms of C. Parvum-induced coagulopathy in mice.

    OpenAIRE

    Mitcheson, H. D.; Hilgard, P.; McCraw, A.; Castro, J E

    1980-01-01

    I.v. injection of Corynebacterium parvum (CP) into C57BL and BALB/c mice caused profound coagulation changes, featuring thrombocytopenia, decreased fibrinogen, increased fibrin/fibrinogen degradation products, and a concomitant microangiopathic haemolytic anaemia. These changes were greatest on the 9th day after CP, with recovery by Day 21. I.p. injection caused similar effects but s.c. injection was ineffective. Radiolabelled-platelet kinetics and distribution after i.v. CP indicated dissemi...

  6. The antitumour activity of maltose tetrapalmitate compared with other immunoadjuvants, and its effectiveness after tumour surgery.

    OpenAIRE

    El Kappany, H.; Chopra, C.; Nigam, V. N.; Brailovsky, C A; Elhilali, M.

    1980-01-01

    The effectivenss of maltose tetrapalmitate (MTP) as an antitumour immune adjuvant was verified by its comparison with other known immunopotentiators, namely BCG, Corynebacterium parvum, levamisole and pyran copolymer. Copenhagen x Fisher 344/CRBL F1 hybrid male rats inoculated s.c. with the Dunning R3327A prostatic adenocarcinoma were used as the test system. All animals treated with immunoadjuvants showed a delay in tumour appearance and inhibition of early tumour growth. MTP was found to be...

  7. Dermoscopy of Pitted Keratolysis

    Directory of Open Access Journals (Sweden)

    Lauren L. Lockwood

    2010-08-01

    Full Text Available Irritated hyperhidrotic soles with multiple small pits are pathognomonic for pitted keratolysis (PK. Here we show the dermatoscopic view of typical pits that can ensure the diagnosis. PK is a plantar infection caused by Gram-positive bacteria, particularly Corynebacterium. Increases in skin surface pH, hyperhidrosis, and prolonged occlusion allow these bacteria to proliferate. The diagnosis is fundamentally clinical and treatment generally consists of a combination of hygienic measures, correcting plantar hyperhidrosis and topical antimicrobials.

  8. Factors influencing the immune response. II. Effects of the physical state of the antigen and of lymphoreticular cell proliferation on the response to intraperitoneal injection of bovine serum albumin in rabbits

    Science.gov (United States)

    Pinckard, R. N.; Weir, D. M.; McBride, W. H.

    1967-01-01

    The injection of Corynebacterium parvum at the same time as centrifuged bovine albumin has been shown not to have the adjuvant effect found when C. parvum is injected 6 days before. The implication of this is discussed and related to mechanisms of antibody synthesis. Whereas particulate alum-precipitated centrifuged bovine albumin was shown to be more effective than centrifuged bovine albumin in inducing primary antibody stimulation, the reverse was true for secondary stimulation by the intraperitoneal route. PMID:6035197

  9. The Effect of Diphtheria Toxin on Nitric Oxide Induction From RAW264.7 Murine Macrophages

    OpenAIRE

    Aybay, Cemalettin

    1998-01-01

    In this study the effect of diphtheria toxin (DT) on nitric oxide (NO) production from RAW 264.7 murine macrophages was investigated. Griess reagent was used to determine NO production by measuring nitrite levels in the culture supernatants. Corynebacterium diphtheriae G12/6 strain-produced DT (Limes flocculation activity and immunodiffusion assays were positive) demonstrated a dose-dependent effect on RAW 264.7 macrophages to induce NO production. L-NAME, an L-arginine analogue, ...

  10. Corynebacterial pneumonia in an African hedgehog.

    Science.gov (United States)

    Raymond, J T; Williams, C; Wu, C C

    1998-04-01

    A 3-mo-old, male African hedgehog (Atelerix albiventris) was anorectic and lethargic for a period of 3 days prior to death. Necropys revealed lungs that were diffusely firm, dark red, and dorsally adhered by fibrinous tags to the pericardial sac. Histopathology revealed necrosuppurative bronchopneumonia with pulmonary abscesses and suppurative pericarditis and myocarditis. A Corynebacterium sp. was isolated from the lungs. We believe this is the first reported case of corynebacterial pneumonia in an African hedgehog. PMID:9577794

  11. Fabrication of Antimicrobial Perspiration Pads and Cotton Cloth Using Amaranthus dubius Mediated Silver Nanoparticles

    OpenAIRE

    M. Jannathul Firdhouse; Lalitha, P.

    2013-01-01

    Silver nanoparticles prepared through a simplistic method using the aqueous extract of Amaranthus dubius were fabricated on perspiration pads and cotton cloth samples to obtain antibacterial textile materials by two different fabrication methods. The antibacterial activity was investigated against the bacteria Corynebacterium which is commonly present in sweat. Silver nanoparticles that serve as antibacterial agents, against pathogenic bacteria, have gained increased applications in medical d...

  12. Xylitol Production From D-Xylose by Facultative Anaerobic Bacteria

    OpenAIRE

    Rangaswamy, Sendil

    2003-01-01

    Seventeen species of facultative anaerobic bacteria belonging to three genera (Serratia, Cellulomonas, and Corynebacterium) were screened for the production of xylitol; a sugar alcohol used as a sweetener in the pharmaceutical and food industries. A chromogenic assay of both solid and liquid cultures showed that 10 of the 17 species screened could grow on D-xylose and produce detectable quantities of xylitol during 24-96 h of fermentation. The ten bacterial species were studied for the effe...

  13. Domain Modeling: NP_110434.2 [SAHG[Archive

    Lifescience Database Archive (English)

    Full Text Available NP_110434.2 chr22 CRYSTAL STRUCTURE OF THE SODIUM-COUPLED GLYCINE BETAINE SYMPORTER... BETP FROM CORYNEBACTERIUM GLUTAMICUM WITH BOUND SUBSTRATE p2wita_ chr22/NP_110434.2/NP_110434.2_holo_16-502.pdb swppa 109N,110I,112V,114S,115A,117I,118L,129E,131I,330L BET 0 ...

  14. Study of molasses / vinasse waste ratio for single cell protein and total microorganisms

    OpenAIRE

    Marcia Luciana Cazetta; Maria Antonia Pedrine Colabone Celligoi

    2006-01-01

    Different molasses/ vinasse ratio were used as substrate to investigate single cell protein and total lipids production by five microorganisms: four yeasts strains: Candida lipolytica, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, a yeast isolated from vinasse lake (denominated LLV98) and a bacterium strain, Corynebacterium glutamicum. The media utilized were: a) 50% molasses and 50% vinasse; b) 25% molasses and 75% vinasse and c) 75% molasses and 25% vinasse. The objective of this work...

  15. Use of sugar cane molasses and vinasse for proteic and lipidic biomass production by yeast and bacteria

    OpenAIRE

    Marcia Luciana Cazetta; Maria Antonia Pedrine Colabone Celligoi

    2005-01-01

    This work evaluated the lipid and protein growth and synthesis capacity by Saccharomyces cerevisiae, Rhodotoruda mucilaginosa, Candida lipolytica, a yeast isolated from vinasse lakes and Corynebacterium glutamicum in 10% molasses and sugar cane crude vinasse. All microorganisms grew both in molasses and vinasse. The highest growth in crude vinasse was performed by R. mucilaginosa (7.05 g/L), and in 10% molasses, by C. lipolytica, yielding 6,09 g/L. In vinasse, the highest protein content in t...

  16. Reduction of Hexavalent Chromium by Viable Cells of Chromium Resistant Bacteria Isolated from Chromite Mining Environment

    OpenAIRE

    Satarupa Dey; Baishali Pandit; A. K. Paul

    2014-01-01

    Environmental contamination of hexavalent chromium [Cr(VI)] is of serious concern for its toxicity as well as mutagenic and carcinogenic effects. Bacterial chromate reduction is a cost-effective technology for detoxification as well as removal of Cr(VI) from polluted environment. Chromium resistant and reducing bacteria, belonging to Arthrobacter, Pseudomonas, and Corynebacterium isolated from chromite mine overburden and seepage samples of Orissa, India, were found to tolerate 12–18 mM Cr(VI...

  17. Identification and antimicrobial susceptibility of bacteria isolated from corneal ulcers of dogs Identificação e susceptibilidade antimicrobiana de bactérias isoladas de úlceras de córnea em cães

    OpenAIRE

    M.R. Prado; E.H.S. Brito; M.D. Girão; J.J.C. Sidrim; Rocha, M. F. G.

    2006-01-01

    A total of 22 clinical specimens were obtained from 19 dogs with corneal ulcer (16 unilateral and three bilateral) for isolation and antimicrobial susceptibility evaluation of the isolated bacteria. Bacterial growth was observed in 100% of the samples (n=22). Staphylococcus intermedius was the predominant species (35.5%), followed by Corynebacterium xerosis (19.3%). Gentamicin, ciprofloxacin, chloramphenicol and tobramycin had a high efficacy against all of the isolated bacteria. The results ...

  18. Dermoscopy of pitted keratolysis

    OpenAIRE

    Lockwood, L L; Gehrke, S. (Susanne); Navarini, A A

    2010-01-01

    Irritated hyperhidrotic soles with multiple small pits are pathognomonic for pitted keratolysis (PK). Here we show the dermatoscopic view of typical pits that can ensure the diagnosis. PK is a plantar infection caused by Gram-positive bacteria, particularly Corynebacterium. Increases in skin surface pH, hyperhidrosis, and prolonged occlusion allow these bacteria to proliferate. The diagnosis is fundamentally clinical and treatment generally consists of a combination of hygienic measures, corr...

  19. IMMUNOBIOLOGY OF DIPHTHERIA. RECENT APPROACHES FOR THE PREVENTION, DIAGNOSIS, AND TREATMENT OF DISEASE

    OpenAIRE

    Kolybo, D.; Labyntsev, A.; Korotkevich, N.; Komisarenko, S.; Romaniuk, S.; Kaberniuk, A.; Oliinyk, O.

    2013-01-01

    Diphtheria is a highly contagious life-threatening disease caused by the toxigenic strains of Corynebacterium diphtheria , which are transformed by a bacteriophage carrying the toxin gene. Diphtheria causative agent and its major virulence factor diphtheria toxin are well studied, but outbreaks of disease still occur worldwide. Rapid development of new methods in immunology and molecular biology is currently leading to improvement of prophylaxis, diagnosis and treatment of diphtheria. This re...

  20. Upper respiratory tract and cutaneous diphtheria

    OpenAIRE

    Prasad, Kishore C.; Kaniyur, Vishnu; Shenoy, Shalini; Prasad, Sampath C.

    2005-01-01

    With the global immunization programme of children there is a progressive decline in the number of diphtheria cases. It is a disease commonly affecting the children caused by Corynebacterium diphtheriae usually affecting the mucous membrane of the nose, pharynx or larynx. Cutaneous diphtheria is a rare entity. We present a rare case of cutaneous diphtheria in a 15-year-old boy with nasal pharyngeal and laryngeal involvement. The patient developed anaphylactic reaction to antidiphtheritic seru...

  1. MODERN APPROACHES IN COMPLEX LABORATORY TESTING FOR DIPHTHERIA

    OpenAIRE

    L. A. Kraeva; E.A. Alekseeva; G. Y. Tseneva; L. A. Lipatova; G. I. Bespalova

    2012-01-01

    Abstract. The criteria of reliable validation of population protection against diphtheria on the base of two indices including quantity of antitoxic antibodies to diphtheria and avidity index have been established. For this purposes it was proposed to use the modified variant of ELISA allowed to detect both indices simultaneously. The formula of probable development of diphtheria in case of the close contact with patients or Corynebacterium diphtheriae bacteria carriers as well as determinati...

  2. Identification of a Human Monoclonal Antibody To Replace Equine Diphtheria Antitoxin for Treatment of Diphtheria Intoxication

    OpenAIRE

    Sevigny, Leila M; Booth, Brian J.; Rowley, Kirk J.; Leav, Brett A.; Cheslock, Peter S.; Kerry A Garrity; Sloan, Susan E.; Thomas, William; Babcock, Gregory J.; Wang, Yang

    2013-01-01

    Diphtheria antitoxin (DAT) has been the cornerstone of the treatment of Corynebacterium diphtheriae infection for more than 100 years. Although the global incidence of diphtheria has declined steadily over the last quarter of the 20th century, the disease remains endemic in many parts of the world, and significant outbreaks still occur. DAT is an equine polyclonal antibody that is not commercially available in the United States and is in short supply globally. A safer, more readily available ...

  3. The life and death of translation elongation factor 2

    DEFF Research Database (Denmark)

    Jørgensen, Rene; Merrill, A.R.; Andersen, Gregers Rom

    2006-01-01

    The eukaryotic elongation factor 2 (eEF2) occupies an essential role in protein synthesis where it catalyses the translocation of the two tRNAs and the mRNA after peptidyl transfer on the 80S ribosome. Recent crystal structures of eEF2 and the cryo-EM reconstruction of its 80S complex now provide...... diphthamide residue, which is ADP-ribosylated by diphtheria toxin from Corynebacterium diphtheriae and exotoxin A from Pseudomonas aeruginosa....

  4. Acute phase protein response in an experimental model of ovine caseous lymphadenitis

    OpenAIRE

    Lang Tamara L; Waterston Mary M; Bence Laura; Lawson Fraser P; Eckersall Peter D; Donachie William; Fontaine Michael C

    2007-01-01

    Abstract Background Caseous lymphadenitis (CLA) is a disease of small ruminants caused by Corynebacterium pseudotuberculosis. The pathogenesis of CLA is a slow process, and produces a chronic rather than an acute disease state. Acute phase proteins (APP) such as haptoglobin (Hp) serum amyloid A (SAA) and α1 acid glycoprotein (AGP) are produced by the liver and released into the circulation in response to pro-inflammatory cytokines. The concentration of Hp in serum increases in experimental CL...

  5. Nasopharyngeal flora in HIV seropositive men who have sex with men.

    OpenAIRE

    Carlin, E M; Hannan, M.; Walsh, J; Talboys, C; Shah, D; Flynn, R.; Azadian, B S; Boag, F C

    1997-01-01

    OBJECTIVES: To assess, in men who were infected with the human immunodeficiency virus (HIV) and who identified themselves as having had sex with men; the nasopharyngeal prevalence of Neisseria gonorrhoeae, N meningitidis, Corynebacterium diphtheriae, and candida species; oral sexual behaviour; the relation between oral flora and oral sexual behavior. METHOD: Nasopharyngeal swabs were taken from HIV seropositive men for culture. The men were also asked to complete a self administered questionn...

  6. Effect of initial pH on mesophilic hydrolysis and acidification of swine manure

    OpenAIRE

    Lin, Lin; Wan, Chunli; Liu, Xiang; Lee, Duu-Jong; Lei, Zhongfang; Zhang, Yi; Hwa, Tay Joo

    2013-01-01

    Effects of initial pH (3–12) on mesophilic hydrolysis and acidification reactions of swine manure was studied. The initial pH changed the microbial community in the suspension so as to affect hydrolysis and acidification reactions on swine manure. At pH 10–12 the Clostridium alkalicellum and/or Corynebacterium humireducens were enriched and the soluble chemical oxygen demand (SCOD), total volatile fatty acids (VFAs), proteins and carbohydrates from manure were increased in quantities. In part...

  7. Development of expression vectors for Escherichia coli based on the pCR2 replicon

    OpenAIRE

    Deb J K; Walia Rupali; Mukherjee K J

    2007-01-01

    Abstract Background Recent developments in metabolic engineering and the need for expanded compatibility required for co-expression studies, underscore the importance of developing new plasmid vectors with properties such as stability and compatibility. Results We utilized the pCR2 replicon of Corynebacterium renale, which harbours multiple plasmids, for constructing a range of expression vectors. Different antibiotic-resistance markers were introduced and the vectors were found to be 100% st...

  8. Caenorhabditis elegans star formation and negative chemotaxis induced by infection with corynebacteria.

    Science.gov (United States)

    Antunes, Camila Azevedo; Clark, Laura; Wanuske, Marie-Therès; Hacker, Elena; Ott, Lisa; Simpson-Louredo, Liliane; de Luna, Maria das Gracas; Hirata, Raphael; Mattos-Guaraldi, Ana Luíza; Hodgkin, Jonathan; Burkovski, Andreas

    2016-01-01

    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 modes of pathogenicity. Depending on the nematode pathogen, aggregates of worms, termed worm-stars, can be formed, or severe rectal swelling, so-called Dar formation, can be induced. Using the human and animal pathogens Corynebacterium diphtheriae and Corynebacterium ulcerans as well as the non-pathogenic species Corynebacterium glutamicum, we show that these coryneform bacteria can also induce star formation slowly in worms, as well as a severe tail-swelling phenotype. While C. glutamicum had a significant, but minor influence on survival of C. elegans, nematodes were killed after infection with C. diphtheriae and C. ulcerans. The two pathogenic species were avoided by the nematodes and induced aversive learning in C. elegans. PMID:26490043

  9. Use of prostaglandin I2 analog in treatment of massive hepatic necrosis associated with endothelial cell injury and diffuse sinusoidal fibrin deposition.

    Science.gov (United States)

    Fujiwara, K; Mochida, S; Ohno, A; Arai, M; Matsui, A; Masaki, N; Hirata, K; Tomiya, T; Yamaoka, M; Nagoshi, S

    1995-01-01

    Endothelial cell damage causes massive hepatic necrosis as a result of fibrin deposition in the hepatic sinusoids. When a stable analog of prostaglandin I2, beraprost sodium, was administered to rats given either dimethylnitrosamine, carbon tetrachloride, or endotoxin following Corynebacterium parvum administration, the hepatic necrosis produced in each was attenuated, but to a greater extent in the dimethylnitrosamine and endotoxin/Corynebacterium parvum models, where fibrin deposition in the hepatic sinusoids occurs, as compared to the carbon tetrachloride model, where such fibrin deposition does not occur. Beraprost sodium reduced the expected increase of portal venous pressure in the endotoxin/Corynebacterium parvum model without affecting plasma thrombin-antithrombin III complex levels. Beraprost sodium also significantly reduced cell killing of both isolated rat hepatocytes and hepatic sinusoidal endothelial cells exposed to tert-butyl hydroperoxide when compared to controls. Beraprost sodium could prove to be a therapeutic candidate for the treatment of hepatic necrosis, particularly in cases associated with fibrin deposition in the hepatic sinusoids because of its fibrin clot-clearing action. PMID:7821117

  10. Improvement of phenolic antioxidants and quality characteristics of virgin olive oil with the addition of enzymes and nitrogen during olive paste processing

    Energy Technology Data Exchange (ETDEWEB)

    Inconomou, D.; Arapoglou, D.; Israilides, C.

    2010-07-01

    The evolution of phenolic compounds and their contribution to the quality characteristics in virgin olive oil during fruit processing was studied with the addition of a combination of various commercial enzymes containing pectinases, polygalacturonases, cellulase and {beta}-glucanase with or without nitrogen flush. Olive fruits (Olea europaea, L.) of the cultivar Megaritiki, at the semi black pigmentation stage of maturity, were used in a 3-phase extraction system in an experiment at industrial scale. The addition of enzymes in the olive paste during processing increased the total phenol and ortho-diphenol contents, as well as some simple phenolic compounds (3,4-DHPEA, p-HPEA) and the secoiridoid derivatives (3,4-DHPEA-EDA and 3,4-DHPEAEA) in olive oil and therefore improved its oxidative stability. Furthermore, enzyme treatment ameliorated the quality parameters of the produced olive oil (acidity and peroxide value) and their sensory attributes. The use of additional N{sub 2} flush with the enzyme treatments did not improve the quality parameters of olive oil any further; however it did not affect the concentration of individual and total sterols or most of the fatty acid composition. Consequently, olive paste treatment with enzymes not only improved the quality characteristics of olive oil and enhanced the overall organoleptic quality, but also increased the olive oil yield. (Author) 33 refs.

  11. Actinoalloteichus nanshanensis sp. nov., isolated from the rhizosphere of a fig tree (Ficus religiosa).

    Science.gov (United States)

    Xiang, Wensheng; Liu, Chongxi; Wang, Xiangjing; Du, Jing; Xi, Lijun; Huang, Ying

    2011-05-01

    A Gram-positive, aerobic actinomycete, designated strain NEAU 119(T), was isolated from the rhizosphere of a fig tree and was characterized using a polyphasic approach. The isolate formed branching, non-fragmenting vegetative hyphae and produced black pigment on yeast extract/malt extract (ISP medium 2). The G+C content of the DNA was 76.6 mol%. The organism had chemotaxonomic characteristics typical of the genus Actinoalloteichus and was closely related to the type strains of Actinoalloteichus cyanogriseus, Actinoalloteichus spitiensis and Actinoalloteichus hymeniacidonis, currently the only three recognized species of the genus Actinoalloteichus, sharing 16S rRNA gene similarities of 96.4, 96.6 and 98.1 %, respectively. However, the results of DNA-DNA hybridization studies demonstrated that the novel strain showed only 46.8 % relatedness with the type strain of A. hymeniacidonis. In addition, a set of phenotypic characteristics also readily distinguished strain NEAU 119(T) from the type strains of recognized species of the genus Actinoalloteichus. According to the above data, it is proposed that strain NEAU 119(T) represents a novel species, Actinoalloteichus nanshanensis sp. nov. The type strain of Actinoalloteichus nanshanensis is NEAU 119(T) ( = CGMCC 4.5714(T) = NBRC 106685(T)). PMID:20562245

  12. Inscriptions on the Interior of the 30th Dynasty Coffin of Nefer-renepet from Akhmim

    Directory of Open Access Journals (Sweden)

    Branislav Anđelković

    2016-02-01

    Full Text Available The anthropoid wooden coffin with plinth (L. 183.5 cm, datable to the mid–4th century B.C. (30th Dynasty, names Nefer-renepet, a dancer of Min from Akhmim. This object represents one of the artistically and technically superior coffins produced by Late Period Egyptian coffin workshops. It was formerly part of the Amherst collection, and was purchased by Ernest Brummer at a Sotheby, Wilkinson & Hodge auction in London in 1921, then donated the same year to the National Museum in Belgrade. The interior of the lid is distinguished by a remarkable ‘gliding Nut motif with upward streaming hair’ (an extremely important iconographic element while the interior of the trough is dominated by a line drawing of Imentet wearing a diagonallyveined maat-feather on her head. The interior decoration includes inscriptions written on the side facets. Written hastily in whitish-yellow line on a rough ground of thick black pigment (in contrast to the fine outer decoration of the coffin these barely legible Stundenwachen texts, are nonetheless significant, and are to be identified as abbreviated texts derived from the Book of Day and Book of Night. They are a manifestation of Late Period magical symbolism stemming from New Kingdom funerary compositions. Their presence on the coffin, however hurried, was intended to ease Nefer-renepet’s passage through the netherworld. Since 1992 the coffin of Nefer-renepet is kept in the Archaeological Collection of the Faculty of Philosophy, University of Belgrade.

  13. Characterization of pre-hispanic pigments by modern analytical techniques; Caracterizacion de pigmentos prehispanicos por tecnicas analiticas modernas

    Energy Technology Data Exchange (ETDEWEB)

    Ortega A, M

    2003-07-01

    In this work, the study of mural painting pigments from two archaeological sites (The Great Temple in Mexico city and Cacaxtla) was performed to know their materials composition, identify their structural characteristics and properties by using modern analytical techniques. Blue, ochre, red and black pigments of Mexica culture (1325-1521 a.C. / late Post Classic period); blue, ochre, red, brown, pink, green and white of Olmeca- Xicalanca culture (700-900 a.C. / Epiclassic period) were studied. Data about materials used, technological evolution, mineralogical background, cultural interchange and origin was obtained. Environmental exposition of these paintings since their discovering has produced changes and damage on their materials. Therefore, stability of some pigments has been notorious, ''Maya Blue'' specially presents extraordinary resistance to diluted and concentrated acids and alkalis including boiling condition, acqua regia, solvents, oxidant and reducing agents, moderate heat and biocorrosi6n; for that reason its study was emphasized. ''Maya Blue'' pigment was synthesized in laboratory using the processes described by historic sources (with indigophera suffruticosa leaves and synthetic indigo) up to obtain a stable pigment including acqua regia action. Clay matrix sorbs nearly 0.4 weight percent of organic dye, which cover 79% of palygorskita surface area. (Author)

  14. [Case report of a patient with ochronosis and arthroplasty of the hip and both knees].

    Science.gov (United States)

    Moslavac, Aleksandra; Moslavac, Sasa; Cop, Renata

    2003-01-01

    Alkaptonuria is a rare hereditary metabolic disorder characterised by absence of the enzyme homogentisic acid oxidase. As a result of this defect homogentisic acid accumulates and is excreted in the urine. The term ochronosis is used to describe bluish-black pigmentation of connective tissue. Ochronotic arthropathy results from the pigmented deposits in the joints of the appendicular and axial skeleton. Findings simulate those of uncomplicated degenerative joint disease, with effusion, articular space narrowing, and bony sclerosis. Our patient is a 70-year old male with ochronotic arthropathy. He has typical ears and sclera discoloration, and had arthroplasty of knees 7 and 4 years ago, respectively. In year 2002, he had undergone total right hip arthroplasty and has been admitted for rehabilitation 14th postoperative day. Individually designed rehabilitation regimen included kinesitherapy, hydrokinesitherapy, and ambulation training with gradual increase in weight bearing exercises and electro-analgesia of associated low back pain. In course of rehabilitation our patient improved his endurance with satisfying range of motion of right hip (flexion 90 degrees, abduction 40 degrees) and strength of hip and thigh musculature. The patient was able to walk with crutches without limitation. We conclude that joint destruction followed by painful locomotion due to ochronotic arthropathy is best treated by total joint arthroplasty, as described in our patient.

  15. POTENTIAL USE OF COLLAGEN HYDROLYSATES FROM CHAMOIS LEATHER WASTE AS INGREDIENT IN LEATHER FINISHING FORMULATIONS

    Directory of Open Access Journals (Sweden)

    POPA Emil

    2016-05-01

    Full Text Available The aim of this paper is the obtaining of value-added products from the dust resulted from chamois leather buffering, a solid waste that raises serious disposal problems, due to its physical state and complex chemical composition. Starting from leather waste, an alkaline hydrolysis was performed followed by the chemical modification of the polypeptyde hydrolysate by polycondensation with dispersions of copolymers of vinyl acetate with acrylic esters and reticulation with glutaraldehyde in order to improve its hydrophobicity. The resulted product can be used/was tested as an ingredient in leather finishing formulations, as binder or carrier agent. In this paper, new finishing mixtures were prepared using pigments and obtained polypeptide hydrolysates as a substitute for casein in pigment pastes. By this method, there were obtained two experimental variants of brown and black pigment pastes which were compared to the pigment pastes with casein binder. Natural grain Box bovine leather samples coated with such admixtures were subjected to physico-mechanical resistance tests, in accordance with the standardized methods. Specific tests carried on finished leather – tensile strength, tear resistance, resistance to grain cracking, dry and wet rubbing fastness, flexural fatigue strength test, etc – showed values of this characteristics comparable to those obtained with casein conventional finishing.

  16. Molecular Study of nifH1, nifH2, nifH3, nifU, nifV, VF Genes and Classical Approach Cared out to Identification of Azotobacter chrococcum from Soil

    Directory of Open Access Journals (Sweden)

    Adel Kamal Khider

    2012-09-01

    Full Text Available The present study aimed to compare classical approach with molecular based method for identification of Azotobacter chrococcum from soil samples. A. chrococcum was isolated from soil source in Erbil city, Iraq. They were cultivated under laboratory conditions using Nitrogen free Azotobacter specific medium. A. chrococcum was present in all soil samples. result shows that A. chrococcum were rod shape, motility occur through the use of peritrichous flagella, cysts-forming, positive to oxidase, catalase and tryptophanase test, unable to liquefy gelatin, with insoluble brown or brown-black pigmentation and darken with age. Utilized starch, sucrose, mannitol and moloanat, but not rhamnose. molecular method based on detection of nifgenes have been successfully applied to describe A. chrococcum isolated from soil. The PCR products for nifH1 1102bp, nifH2 246bp, nifH3 128bp, nifU 930bp, nifV 1146bp and VF gene 594bp were detected on gel electrophoresis, while no bands observed for negative control. The isolated bacteria considered Azotobacte chrococcum belonging to Genus Azotobacter.

  17. Genetic Factors and Host Traits Predict Spore Morphology for a Butterfly Pathogen

    Directory of Open Access Journals (Sweden)

    Jacobus C. de Roode

    2013-08-01

    Full Text Available Monarch butterflies (Danaus plexippus throughout the world are commonly infected by the specialist pathogen Ophryocystis elektroscirrha (OE. This protozoan is transmitted when larvae ingest infectious stages (spores scattered onto host plant leaves by infected adults. Parasites replicate internally during larval and pupal stages, and adult monarchs emerge covered with millions of dormant spores on the outsides of their bodies. Across multiple monarch populations, OE varies in prevalence and virulence. Here, we examined geographic and genetic variation in OE spore morphology using clonal parasite lineages derived from each of four host populations (eastern and western North America, South Florida and Hawaii. Spores were harvested from experimentally inoculated, captive-reared adult monarchs. Using light microscopy and digital image analysis, we measured the size, shape and color of 30 replicate spores per host. Analyses examined predictors of spore morphology, including parasite source population and clone, parasite load, and the following host traits: family line, sex, wing area, and wing color (orange and black pigmentation. Results showed significant differences in spore size and shape among parasite clones, suggesting genetic determinants of morphological variation. Spore size also increased with monarch wing size, and monarchs with larger and darker orange wings tended to have darker colored spores, consistent with the idea that parasite development depends on variation in host quality and resources. We found no evidence for effects of source population on variation in spore morphology. Collectively, these results provide support for heritable variation in spore morphology and a role for host traits in affecting parasite development.

  18. Black Molecular Adsorber Coatings for Spaceflight Applications

    Science.gov (United States)

    Abraham, Nithin Susan; Hasegawa, Mark Makoto; Straka, Sharon A.

    2014-01-01

    The molecular adsorber coating is a new technology that was developed to mitigate the risk of on-orbit molecular contamination on spaceflight missions. The application of this coating would be ideal near highly sensitive, interior surfaces and instruments that are negatively impacted by outgassed molecules from materials, such as plastics, adhesives, lubricants, epoxies, and other similar compounds. This current, sprayable paint technology is comprised of inorganic white materials made from highly porous zeolite. In addition to good adhesion performance, thermal stability, and adsorptive capability, the molecular adsorber coating offers favorable thermal control characteristics. However, low reflectivity properties, which are typically offered by black thermal control coatings, are desired for some spaceflight applications. For example, black coatings are used on interior surfaces, in particular, on instrument baffles for optical stray light control. Similarly, they are also used within light paths between optical systems, such as telescopes, to absorb light. Recent efforts have been made to transform the white molecular adsorber coating into a black coating with similar adsorptive properties. This result is achieved by optimizing the current formulation with black pigments, while still maintaining its adsorption capability for outgassing control. Different binder to pigment ratios, coating thicknesses, and spray application techniques were explored to develop a black version of the molecular adsorber coating. During the development process, coating performance and adsorption characteristics were studied. The preliminary work performed on black molecular adsorber coatings thus far is very promising. Continued development and testing is necessary for its use on future contamination sensitive spaceflight missions.

  19. Sexually dimorphic body color is regulated by sex-specific expression of yellow gene in ponerine ant, Diacamma sp.

    Directory of Open Access Journals (Sweden)

    Satoshi Miyazaki

    Full Text Available Most hymenopteran species exhibit conspicuous sexual dimorphism due to ecological differences between the sexes. As hymenopteran genomes, under the haplodiploid genetic system, exhibit quantitative differences between sexes while remaining qualitatively identical, sexual phenotypes are assumed to be expressed through sex-specific gene usage. In the present study, the molecular basis for expression of sexual dimorphism in a queenless ant, Diacamma sp., which exhibits a distinct color dimorphism, was examined. Worker females of the species appear bluish-black, while winged males exhibit a yellowish-brown body color. Initially, observations of the pigmentation processes during pupal development revealed that black pigmentation was present in female pupae but not in males, suggesting that sex-specific melanin synthesis was responsible for the observed color dimorphism. Therefore, five orthologs of the genes involved in the insect melanin synthesis (yellow, ebony, tan, pale and dopa decarboxylase were subcloned and their spatiotemporal expression patterns were examined using real-time quantitative RT-PCR. Of the genes examined, yellow, which plays a role in black melanin synthesis in insects, was expressed at higher levels in females than in males throughout the entire body during the pupal stage. RNA interference of yellow was then carried out in order to determine the gene function, and produced females with a more yellowish, brighter body color similar to that of males. It was concluded that transcriptional regulation of yellow was responsible for the sexual color dimorphism observed in this species.

  20. Hemosiderotic clear-cell acanthoma: A pigmented mimicker

    Directory of Open Access Journals (Sweden)

    Leonardo Bugatti

    2011-01-01

    Full Text Available The authors report on a case of a 65-year-old man with pigmented clear-cell acanthoma located on the right thigh. Dermoscopy disclosed a peculiar picture consisting of diffuse black pigmentation with a superficial greyish veil in the central portion, dotted-to-globular dark red-black structures mainly located at the periphery with a homogenous regular reticular arrangement; peripheral translucid desquamation. Dermoscopic features are correlated with the histology, where hemosiderin deposits present in a sheet-like arrangement in the perivascular papillary dermis and in a band-like disposition in the reticular dermis at the base of the lesion can account for the pigmented picture. The lesion arose on a trauma-prone skin site; thus the authors believe that traumatic irritation may be responsible for the clinical and dermoscopic pictures, giving rise to a reaction similar in a way to the Auspitz′s sign provocated by trauma for psoriasis. Red blood cells extravasation from extremely superficialized capillaries may have led to hemosiderin deposition in the papillary and the reticular dermis.

  1. A possible dominant white gene in Jersey cattle

    Directory of Open Access Journals (Sweden)

    Sponenberg D Phillip

    2001-01-01

    Full Text Available Abstract A white heifer ("Snow" was born in 1991 from coloured registered Jersey parents. She produced six calves sired by coloured Jersey bulls: three white bull calves, two white heifer calves, and one coloured bull calf. One of the white bull calves was mated with 40 Hereford × Friesian yearling heifers (white face, predominantly black body with some white patches. The 38 resulting calves included 16 white and 22 coloured calves. Twelve of the 16 white calves were heifers and four were bulls. Red or black spotting was recorded on some white calves. The results are consistent with an autosomal dominant mutant causing the white phenotype. The mutation appears to have arisen spontaneously in Snow, then passing to her white progeny and white grand-progeny. The white individuals varied from entirely white in a few cases, to most having some residual small areas of red or black pigmentation in patterns not typical of other reported white spotting patterns of cattle.

  2. Nondestructive Raman investigation on wall paintings at Sala Vaccarini in Catania (Sicily)

    Science.gov (United States)

    Barone, Germana; Bersani, Danilo; Coccato, Alessia; Lauwers, Debbie; Mazzoleni, Paolo; Raneri, Simona; Vandenabeele, Peter; Manzini, Davide; Agostino, Giuseppe; Neri, Nicola Francesco

    2016-09-01

    In this work, the results of a Raman campaign for studying seventeenth-century Sicilian frescoes, by using two portable Raman systems, equipped with different excitation sources (785 and 1064 nm), are proposed. The measurements were performed with the aim to provide an in situ diagnostic analysis of the wall paintings (in terms of colorants and preparation layer) and to support the conservators in the framework of the ongoing restoration. The combined use of the two Raman spectrometers has given a complete overview on the artist palette and on the state of preservation of frescoes, also informing us about the technique employed by the painter. Natural pigments as hematite, vermillion, goethite, lead red, lead white and carbon-based black pigments have been identified. Additionally, the application of a transitional Romanesque-Renaissance frescoes method has been noticed by the systematic combined presence of calcite and gypsum in the substrate. Finally, the analyses have highlighted the presence of degradation products, mainly related to alteration of lead-based pigments.

  3. Raman microscopy of hand stencils rock art from the Yabrai Mountain, Inner Mongolia Autonomous Region, China

    Science.gov (United States)

    Hernanz, Antonio; Chang, Jinlong; Iriarte, Mercedes; Gavira-Vallejo, Jose M.; de Balbín-Behrmann, Rodrigo; Bueno-Ramírez, Primitiva; Maroto-Valiente, Angel

    2016-07-01

    A series of rock art pictographs in the form of hand stencils discovered in two sites of the Yabrai Mountain, Inner Mongolia Autonomous Region (China) has been studied by micro-Raman spectroscopy, X-ray photoelectron spectroscopy and scanning electronic microscopy combined with energy dispersive X-ray spectroscopy for the first time. These studies have made possible to characterise the materials present. The minerals α-quartz, phlogopite, albite and microcline have been identified in the granitic rocks supporting the paintings. Calcite and dolomite micro-particles detected on the rock surface have been attributed to desert dust. Accretions of gypsum, anhydrite and whewellite have also been identified on the rock surface. Haematite is the pigment used in the red pictographs, whereas well-crystallised graphite has been used in the black ones. The use of crystalline graphite instead of amorphous carbon (charcoal, soot or bone black) as a black pigment in rock art is an interesting novelty. Overlapped hands are proposed as a new type of hand stencils to make an unusual pictorial symbol in rock art that has been found in these sites.

  4. Characterization of pre-hispanic pigments by modern analytical techniques

    International Nuclear Information System (INIS)

    In this work, the study of mural painting pigments from two archaeological sites (The Great Temple in Mexico city and Cacaxtla) was performed to know their materials composition, identify their structural characteristics and properties by using modern analytical techniques. Blue, ochre, red and black pigments of Mexica culture (1325-1521 a.C. / late Post Classic period); blue, ochre, red, brown, pink, green and white of Olmeca- Xicalanca culture (700-900 a.C. / Epiclassic period) were studied. Data about materials used, technological evolution, mineralogical background, cultural interchange and origin was obtained. Environmental exposition of these paintings since their discovering has produced changes and damage on their materials. Therefore, stability of some pigments has been notorious, ''Maya Blue'' specially presents extraordinary resistance to diluted and concentrated acids and alkalis including boiling condition, acqua regia, solvents, oxidant and reducing agents, moderate heat and biocorrosi6n; for that reason its study was emphasized. ''Maya Blue'' pigment was synthesized in laboratory using the processes described by historic sources (with indigophera suffruticosa leaves and synthetic indigo) up to obtain a stable pigment including acqua regia action. Clay matrix sorbs nearly 0.4 weight percent of organic dye, which cover 79% of palygorskita surface area. (Author)

  5. PIXE facility at Centro Atómico Bariloche

    Energy Technology Data Exchange (ETDEWEB)

    Limandri, S. [Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina (CONICET) (Argentina); Olivares, C. [Centro Atómico Bariloche, Comisión Nacional de Energía Atómica, Av. E. Bustillo 9500, San Carlos de Bariloche, Río Negro (Argentina); Rodriguez, L. [Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina (CONICET) (Argentina); Bernardi, G. [Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina (CONICET) (Argentina); Centro Atómico Bariloche, Comisión Nacional de Energía Atómica, Av. E. Bustillo 9500, San Carlos de Bariloche, Río Negro (Argentina); Suárez, S., E-mail: suarez@cab.cnea.gov.ar [Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina (CONICET) (Argentina); Instituto Balseiro, Universidad Nacional de Cuyo, Av. E. Bustillo 9500, San Carlos de Bariloche, Río Negro (Argentina)

    2014-01-01

    A 1.7 MeV tandem accelerator has been recently installed at Centro Atómico Bariloche, Argentina. This communication includes a brief description of the system facility with the main associated equipment, in particular that used for PIXE measurements. The X-ray detector efficiency was experimentally determined by comparing X-ray characteristic spectra with theoretical predictions. The knowledge of this parameter allows us to perform quantification without standards with good precision. We show some recent applications of PIXE in archaeology and forensic science done at our laboratory. In the archaeological field we determined elements constituent of pastes and black, white, brown and ocher pigments from Aguada Portezuelo Culture (600–900 AD). This study allowed us to infer about the presence of psilomelante mineral (a mineral with barium) as precursor of some black pigments. In the forensic field we determined trace elements that are present in seven gunpowders commonly used in Argentina. We note that any gunpowder can be distinguished by the presence of trace elements.

  6. Heavy Water as a Probe of the Free Radical Nature and Electrical Conductivity of Melanin.

    Science.gov (United States)

    Rienecker, Shermiyah B; Mostert, A Bernardus; Schenk, Gerhard; Hanson, Graeme R; Meredith, Paul

    2015-12-01

    Melanins are pigmentary macromolecules found in many locations throughout nature including plants and vertebrate animals. It was recently proposed that the predominant brown-black pigment eumelanin is a mixed ionic-electronic conductor which has led to renewed interest in its basic properties as a model bioelectronic material. This exotic hybrid electrical behavior is strongly dependent upon hydration and is closely related to the free radical content of melanin which is believed to be a mixed population of two species: the semiquinone (SQ) and a carbon-centered radical (CCR). The predominant charge carrier is the proton that is released during the formation of the SQ radical and controlled by a comproportionation equilibrium reaction. In this paper we present a combined solid-state electron paramagnetic resonance (EPR), adsorption, and hydrated conductivity study using D2O as a probe. We make specific predictions as to how the heavy isotope effect, in contrast to H2O, should perturb the comproportionation equilibrium and the related outcome as far as the electrical conductivity is concerned. Our EPR results confirm the proposed two-spin mechanism and clearly demonstrate the power of combining macroscopic measurements with observations from mesoscopic probes for the study of bioelectronic materials.

  7. Raman, SEM-EDS and XRPD investigations on pre-Columbian Central America "estucado" pottery

    Science.gov (United States)

    Casanova Municchia, Annalaura; Micheli, Mario; Ricci, Maria Antonietta; Toledo, Michelle; Bellatreccia, Fabio; Lo Mastro, Sergio; Sodo, Armida

    2016-03-01

    Seventeen different colored fragments from six selected pre-Columbian estucado ceramics from El Salvador have been investigated by Raman spectroscopy, scanning electron microscope coupled to an energy dispersive spectrometer (SEM/EDS) and X-ray powder diffraction (XRPD). The peculiarity of this kind of ceramics consist of the unusual presence of a white engobe, traditionally termed stucco, between the ceramic body and the decoration elements, hence the name estucado ceramics. The aim of this work was to study the unusual manufacturing technique and to identify the chemical composition of the engobe and of the pigment palette. The results showed that the stucco layer is made of clay (kaolinite) with traces of titanium oxide (anatase). Remarkably, this is the same composition of the white pigments used for the decoration layer, thus excluding an early use of natural titanium oxide as a white pigment in the estucado productions as suggested in previous investigations. Moreover, the presence of kaolinite and anatase both in the stucco and in the decoration layer suggests a cold-working or low temperature technique. The red, yellow and green decorations were realized by the use of natural ochre, while in all the blue and gray decorations Maya blue pigment was identified. Finally, an amorphous carbon pigment of vegetal origin and manganese oxide were used to obtain black pigments.

  8. Suitability of the voltammetry of immobilized microparticles to detect and discriminate lead compounds in microsamples of ancient black cosmetics

    International Nuclear Information System (INIS)

    Lead compounds, in the form of galena, have been used as eye cosmetics since ancient times and they still appear in traditional products. The presence of some black pigments in several archaeological glass objects gave us the opportunity to study the nature of these products, in order to evaluate the results of an electrochemical method to characterise lead compounds when only heterogeneous microsamples are available and the material requires to be as less manipulated as possible. The present paper describes the data obtained with an electroanalytical methodology (Voltammetry of immobilised microparticles, VMP), and a complementary Scanning Electron Microscopy study, to detect these lead compounds. A simple and fast characterisation method is discussed in order to be able to monitor the presence of lead compounds without previous sample preparation, directly in the solid micro-sample, and using an affordable instrumentation. PbS could be identified and distinguished from other lead compounds in cosmetics by Differential Pulse VMP in 1 M NaCl medium by a set of three peaks (−0.63 V, −0.44 V, +0.45 V) in the anodic voltammogram and two peaks around −0.65 V and +0.45 V in the cathodic scan. The results showed the suitability of the electroanalytical method, offering valuable information about the chemical and mineral composition of the samples using a minimum amount of material, and illustrating the advantages in terms of time consuming, cost and accessibility of the laboratory facilities

  9. Pyogranulomatous pneumonia in goats caused by an undescribed Porphyromonas species, "Porphyromonas katsikii".

    Science.gov (United States)

    Filioussis, George; Petridou, Evanthia; Karavanis, Emmanouel; Frey, Joachim

    2015-03-01

    A yet-undescribed bacterial species, tentatively named "Porphyromonas katsikii," was isolated from individuals of a small goat herd with pyogranulomatous pneumonia during an outbreak of acute respiratory disease. The isolated bacteria grew in the form of black-pigmented colonies after 14 days of incubation under anaerobic conditions at 37°C on a tryptic soy blood agar medium. The bacteria were identified as a yet-undescribed Porphyromonas species by determination of the nucleotide sequence of the rrs 16S rRNA gene, and this species was tentatively named Porphyromonas katsikii. PCR amplification with specific primers for this yet-undescribed species revealed the presence of P. katsikii in the lung tissue of all affected animals, while no PCR signals were evidenced from the lungs of healthy goats or from goats with pasteurellosis caused by Mannheimia haemolytica. These data indicate P. katsikii as the causative agent of acute respiratory distress. P. katsikii is phylogenetically related to Porphyromonas somerae and Porphyromonas levii, which cause pathologies in humans and animals, respectively. P. katsikii was not detected by PCR from samples of the gingival pockets or of the faces of healthy goats. PMID:25540395

  10. X-ray diffraction and Raman spectroscopy study of white decorations on tricolored ceramics from Northwestern Argentina

    Science.gov (United States)

    Freire, E.; Acevedo, V.; Halac, E. B.; Polla, G.; López, M.; Reinoso, M.

    2016-03-01

    White virgules, commas, and dot designs on tricolored ceramics are sporadically found in different archaeological sites located in Northwestern Argentina area, as Puna and Quebrada de Humahuaca. This decorating style has been reported in several articles, but few previous archaeometric studies have been carried out on the pigment composition. Fragments from Puna and Quebrada archaeological sites, belonging to Regional Development Period (900-1430 AD), were analyzed by X-ray diffraction and Raman spectroscopy in order to characterize the pigments employed. Red and black pigments are based on iron and manganese oxides, as it has been extensively reported for the NW Argentina area. White pigments from white virgules, comma, and dot designs have shown different composition. Hydroxyapatite was found in samples from Doncellas site (North Puna region), and calcium and calcium-magnesium containing compounds, as vaterite and dolomite, along with titanium containing compounds were detected on samples from Abralaite (Central Puna region) and Gasoducto (Quebrada de Humahuaca region). It has been concluded that pigment composition is not characteristic of a unique region.

  11. Rapamycin treatment causes developmental delay, pigmentation defects, and gastrointestinal malformation on Xenopus embryogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Moriyama, Yuki [Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529 (Japan); Ohata, Yoshihisa [Department of Education (Sciences), Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529 (Japan); Mori, Shoko [Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529 (Japan); Matsukawa, Shinya [Department of Education (Sciences), Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529 (Japan); Michiue, Tatsuo [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902 (Japan); Asashima, Makoto [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902 (Japan); Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Baien, Tsukuba, Ibaraki 305-8562 (Japan); Kuroda, Hiroki, E-mail: ehkurod@ipc.shizuoka.ac.jp [Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529 (Japan); Department of Education (Sciences), Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529 (Japan)

    2011-01-28

    Research highlights: {yields} Does famous anti-aging drug rapamycin work from the beginning of life? The answer is yes. {yields} This study shows that developmental speed of frog embryo was dose-dependently decreased by rapamycin treatment. {yields} In additions, morphogenetic effects such as less pigmentations and gut malformation are occurred by rapamycin. -- Abstract: Rapamycin is a drug working as an inhibitor of the TOR (target of rapamycin) signaling pathway and influences various life phenomena such as cell growth, proliferation, and life span extension in eukaryote. However, the extent to which rapamycin controls early developmental events of amphibians remains to be understood. Here we report an examination of rapamycin effects during Xenopus early development, followed by a confirmation of suppression of TOR downstream kinase S6K by rapamycin treatment. First, we found that developmental speed was declined in dose-dependent manner of rapamycin. Second, black pigment spots located at dorsal and lateral skin in tadpoles were reduced by rapamycin treatment. Moreover, in tadpole stages severe gastrointestinal malformations were observed in rapamycin-treated embryos. Taken together with these results, we conclude that treatment of the drug rapamycin causes enormous influences on early developmental period.

  12. Micro-Raman analysis of the pigments on painted pottery figurines from two tombs of the Northern Wei Dynasty in Luoyang

    Science.gov (United States)

    Liu, Zhaojun; Han, Yunxia; Han, Ligang; Cheng, Yongjian; Ma, Yiqiang; Fang, Li

    2013-05-01

    The pigments on the painted pottery figurines from two tombs of Northern Wei Dynasty (AD 386-534) in Luoyang were analyzed by Raman microscopy. All the pigments were identified compared with the Raman spectra of standard pigments. The red pigments were identified as haematite, the blue pigment as lapis lazuli, the green pigment as malachite, the black pigment as carbon black and the white pigment as calcite. Similar pigments were used in the two tombs despite the pottery figurines were very different in artistic style. The use of lapis lazuli as blue pigment on Chinese painted pottery figurines was found for the first time. This pigment and the painted pottery figurine of Sogdians are of great archaeological significance because it demonstrated that the trade and cultural exchanges via the Silk Road had extended to Luoyang city in the Northern Wei Dynasty. The result also confirms that micro-Raman spectroscopy is a powerful analytical method for the identification of pigments on ancient artworks.

  13. Study on High Hiding Power Calcium Carbonate/TiO2 Composite White Pigment%高遮盖力碳酸钙/钛白粉复合白色颜料研究

    Institute of Scientific and Technical Information of China (English)

    王岩岩; 张俭; 盛嘉伟

    2013-01-01

    The calcium carbonate/TiO2 (C/TCP) was prepared by mechano-chemical method. The hiding power of C/TCP was improved by adding a small amount of high hiding power iron oxide red and iron oxide black pigments. The influences of additive amount of iron oxide red and iron oxide black to the properties of C/TCP were studied. The results showed that the hiding power of C/TCP was 21.1g/m2, whiteness was 83.4%, oil absorption was 12.8 g/100g, while the iron oxide black content was 0.08%.%采用机械力化学法制备了碳酸钙/TiO2复合粉体(C/TCP),通过添加微量高遮盖力的氧化铁红、氧化铁黑颜料提高了复合粉体的遮盖力,研究了氧化铁红、氧化铁黑添加量对复合粉体性能的影响。结果表明,当氧化铁黑含量为0.08%时,复合粉体的遮盖力为21.1 g/m2、白度为83.4%、吸油量为12.8 g/100 g。

  14. Lasiodiplodia theobromae keratitis: a case report and review of literature.

    Science.gov (United States)

    Saha, Suman; Sengupta, Jayangshu; Banerjee, Debdulal; Khetan, Archana

    2012-10-01

    A case report and review of literature is reported of a rare case of fungal keratitis from eastern India. A 32-year-old woman with a history of vegetative trauma presented with keratitis in left eye. Microbiological examination of corneal scraping showed refractile hyphae with aseptate branching filaments and black pigmented colonies on multiple solid agar medium. Organism was identified from culture using D1/D2 region of LSU (Large Sub Unit: 28S rDNA)-based molecular technique. PCR amplified a band with a sequence that was 100 % homologous with Lasiodiplodia theobromae. The organism was susceptible to amphotericin B and voriconazole and demonstrated resistance to itraconazole and fluconazole. A therapeutic keratoplasty was performed following non-responsiveness to initial topical voriconazole (2 %) therapy. Recurrence in graft was controlled with topical voriconazole and intracameral amphotericin B. However, the graft failed at the end of 3 months. L. theobromae is a rare cause of fungal keratitis. Management of these cases is difficult, often involving surgical procedures. PMID:22544631

  15. Micro-Raman analysis of the pigments on painted pottery figurines from two tombs of the Northern Wei Dynasty in Luoyang.

    Science.gov (United States)

    Liu, Zhaojun; Han, Yunxia; Han, Ligang; Cheng, Yongjian; Ma, Yiqiang; Fang, Li

    2013-05-15

    The pigments on the painted pottery figurines from two tombs of Northern Wei Dynasty (AD 386-534) in Luoyang were analyzed by Raman microscopy. All the pigments were identified compared with the Raman spectra of standard pigments. The red pigments were identified as haematite, the blue pigment as lapis lazuli, the green pigment as malachite, the black pigment as carbon black and the white pigment as calcite. Similar pigments were used in the two tombs despite the pottery figurines were very different in artistic style. The use of lapis lazuli as blue pigment on Chinese painted pottery figurines was found for the first time. This pigment and the painted pottery figurine of Sogdians are of great archaeological significance because it demonstrated that the trade and cultural exchanges via the Silk Road had extended to Luoyang city in the Northern Wei Dynasty. The result also confirms that micro-Raman spectroscopy is a powerful analytical method for the identification of pigments on ancient artworks. PMID:23501716

  16. Erythristic leopards Panthera pardus in South Africa

    Directory of Open Access Journals (Sweden)

    Tara J. Pirie

    2016-05-01

    Full Text Available Background: Leopards (Panthera pardus show genetically determined colour variation. Erythristic (strawberry morphs, where individuals are paler and black pigment in the coat is replaced by a red-brown colour, are exceptionally rare in the wild. Historically, few records exist, with only five putative records known from India.Objectives: To record the presence of erythristic leopards in our study site (Thaba Tholo Wilderness Reserve, Mpumalanga and to collate records from across South Africa. Method: A network of camera traps was used to record individual leopards at Thaba Tholo. We also surveyed local experts, searched the popular South African press, and used social media to request observations.Results: Two out of 28 individual leopards (7.1% recorded in our study site over 3 years were of this colour morph. We obtained records of five other erythristic leopards in the North West and Mpumalanga regions, with no reports outside of this population.Conclusions: Erythristic leopards are widely dispersed across north-east South Africa, predominantly in the Lydenburg region, Mpumalanga. The presence of this rare colour morph may reflect the consequences of population fragmentation.

  17. Mediaeval cantorals in the Valladolid Biblioteca: FT-Raman spectroscopic study.

    Science.gov (United States)

    Edwards, H G; Farwell, D W; Rull Perez, F; Medina Garcia, J

    2001-03-01

    Raman spectroscopic studies of three mediaeval cantorals in the Biblioteca of the University of Valladolid has revealed information about the pigments used on these large manuscripts. Although executed in a simple colour palette, very pure cinnabar was used as the major colourant, offsetting the carbon black of the verses and script. A dark blue colour was achieved using a mixture of azurite (basic copper carbonate) and carbon, whereas a light blue colour was azurite alone. A grey colour was achieved using azurite, carbon particles and a calcareous 'limewash'. A yellow pigment, used sparely in the cantorals was ascribed to saffron; unusually, there was no evidence for the presence of the yellow mineral pigments orpiment, realgar and massicot. In several regions of the vellum specimens, evidence for biodeterioration was observed through the signatures of hydrated calcium oxalate. We report for the first time the Raman spectra of pigment in situ on a vellum fragment, which also shows evidence of substrate bands; comparison of black and red pigmented regions of vellum specimens has shown the presence of calcium oxalate in the black pigmented script but not in the red pigment regions, which suggests that the cinnabar in the red-pigmented regions acts as a toxic protectant for the vellum substrate against biological colonisation processes.

  18. Microbial Tyrosinases: Promising Enzymes for Pharmaceutical, Food Bioprocessing, and Environmental Industry

    Directory of Open Access Journals (Sweden)

    Kamal Uddin Zaidi

    2014-01-01

    Full Text Available Tyrosinase is a natural enzyme and is often purified to only a low degree and it is involved in a variety of functions which mainly catalyse the o-hydroxylation of monophenols into their corresponding o-diphenols and the oxidation of o-diphenols to o-quinones using molecular oxygen, which then polymerizes to form brown or black pigments. The synthesis of o-diphenols is a potentially valuable catalytic ability and thus tyrosinase has attracted a lot of attention with respect to industrial applications. In environmental technology it is used for the detoxification of phenol-containing wastewaters and contaminated soils, as biosensors for phenol monitoring, and for the production of L-DOPA in pharmaceutical industries, and is also used in cosmetic and food industries as important catalytic enzyme. Melanin pigment synthesized by tyrosinase has found applications for protection against radiation cation exchangers, drug carriers, antioxidants, antiviral agents, or immunogen. The recombinant V. spinosum tryosinase protein can be used to produce tailor-made melanin and other polyphenolic materials using various phenols and catechols as starting materials. This review compiles the recent data on biochemical and molecular properties of microbial tyrosinases, underlining their importance in the industrial use of these enzymes. After that, their most promising applications in pharmaceutical, food processing, and environmental fields are presented.

  19. Microbial tyrosinases: promising enzymes for pharmaceutical, food bioprocessing, and environmental industry.

    Science.gov (United States)

    Zaidi, Kamal Uddin; Ali, Ayesha S; Ali, Sharique A; Naaz, Ishrat

    2014-01-01

    Tyrosinase is a natural enzyme and is often purified to only a low degree and it is involved in a variety of functions which mainly catalyse the o-hydroxylation of monophenols into their corresponding o-diphenols and the oxidation of o-diphenols to o-quinones using molecular oxygen, which then polymerizes to form brown or black pigments. The synthesis of o-diphenols is a potentially valuable catalytic ability and thus tyrosinase has attracted a lot of attention with respect to industrial applications. In environmental technology it is used for the detoxification of phenol-containing wastewaters and contaminated soils, as biosensors for phenol monitoring, and for the production of L-DOPA in pharmaceutical industries, and is also used in cosmetic and food industries as important catalytic enzyme. Melanin pigment synthesized by tyrosinase has found applications for protection against radiation cation exchangers, drug carriers, antioxidants, antiviral agents, or immunogen. The recombinant V. spinosum tryosinase protein can be used to produce tailor-made melanin and other polyphenolic materials using various phenols and catechols as starting materials. This review compiles the recent data on biochemical and molecular properties of microbial tyrosinases, underlining their importance in the industrial use of these enzymes. After that, their most promising applications in pharmaceutical, food processing, and environmental fields are presented.

  20. 糖尿病酮症酸中毒患者口腔菌群改变的相关因素研究%Study on the changes of oral flora in patients with diabetic ketoacidosis

    Institute of Scientific and Technical Information of China (English)

    余秀颜; 刘平; 窦宇红; 贝奕冰; 华志英; 张旭芳

    2011-01-01

    Objective To study the changes of oral flora in patients with diabetic ketoacidosis,in order to provide evidence for making oral nursing intervention and hygiene education. Methods 35 patients with diabetic ketoacidosis were named as group A,35 non-diabetic patients with chronic periodontitis (diagnostic criteria:periodontal pocket 14 mm) were named as group B,35 nonketotic patients with diabetes mellitus were named as group C,then all the patients were detected for the oral disease and the oral hygiene was evaluated. Results The gingival index,plaque index,tooth mobility,probing depth and hemorrhage after the detection of three groups had no significant differences. The detection rate of streptococcus oralis, lactobacillus,fusobacterium nucleatum,black-pigment bacteria,Capnocytophaga gingivalis,actinomycetes, escherichia coli,staphylococcus aureus and pseudomonas aeruginosa had no significant differences,there was a positive correlation between quantity of black-pigment bacteria, Capnocytophaga gingivalis and fasting blood glucose and glycosylated hemoglobin of patients with diabetic ketoacidosis. Conclusions Diabetic ketoacidosis strengthened the bacterial invasion and oral colonization of patients.%目的 探讨糖尿病酮症酸中毒患者口腔菌群的改变,为糖尿病患者制订口腔护理干预和口腔卫生宣教提供理论依据.方法 糖尿病酮症酸中毒患者35例(A组),同时选择35例非糖尿病慢性牙周炎(诊断标准:患牙牙周袋>14 mm)患者(B组)和35例糖尿病非酮症患者(C组)作为对照组,检查记录口腔疾病状况,评估口腔卫生状况.结果 3组牙龈指数、菌斑指数、牙齿松动、探诊深度、探诊后出血情况比较,均无显著差异;3组口腔链球菌、口腔乳杆菌、核梭杆菌、产黑菌、二氧化碳噬纤维菌无显著差异,放线菌、大肠埃希菌、金黄色葡萄球菌、铜绿假单胞菌、肺炎克雷伯菌、鲍曼复合醋酸钙不动杆菌、白假丝酵母菌