• 제목/요약/키워드: mycolic acids

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Quantitative Analysis of Corynomycolic Acids in Fermentation Broth

  • Jang, Ki-Hyo;Park, Yong-Il;Britz, Margaret-L.
    • Journal of Microbiology and Biotechnology
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    • 제12권5호
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    • pp.793-800
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    • 2002
  • The mycolic acids and fatty acids of mycolic acid- containing bacteria in various types of fluids were analyzed using capillary gas chromatography and mass spectrometry. As model strains, Brevibacterium and Coryebacterium species, which have corynomycolic acids ill the range of $C_{32}C_{36}$ in the whole cell, were investigated. Optimized solvents extraction procedures for the mycolic acids and fatty acids from the culture fluids were: chloroform/methanol (1:2, v/v) as the first extraction solvents fur 4 h; and chlorofunuwater (1:1, v/v) as the second extraction solvents far 1 h. These conditions gave above 95% recovery yields fur mycolic acids from the culture fluids. The mycolic acid profile for the whole cells and the culture fluids were similar fur all the media tested. Thus, the procedure described here could be applied for the identification of mycolic acid-containing bacteria in fermentation broth or liquid from of foods.

Characterization of the Cell-Surface Barriers to Plasmid Transformation in Corynebacterium glutamicum

  • Jang, Ki-Hyo;Paul J. Chambers;Chun, Uck-Han;Margare L.Britz
    • Journal of Microbiology and Biotechnology
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    • 제11권2호
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    • pp.294-301
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    • 2001
  • The effects of including glycine and isonicotinic acid hydrazide (INH) in the growth medium (Luria broth, LBG) on the subsequent lysozyme-imduced protoplast formation and transformation efficiency of Corynebacterium glutamicum were studied. The transformation efficiency of C. glutamicum AS019 increased up to 100-fold as the ocncentrationof glycine in the media increased from 0% to 5% (w/v), relative to cells grown in the absence of glycine. The presence of 5 mg/ml INH in the growth medium led to a further 10-fold increase in transformation efficiency. In addition, this transformation protocol was successfully applied to other strains of C. glutamicum. Both chemicals affected the mycolic acid attachment to the cell surface of C. glutamicum, when INH, the relative percentage of fatty acids of AS019 to the total lipids (mycolic acid plus fatty acids) decreased from 76.9% (in LBG) to 72.9% (in LBG-2% glycine) and 66.4% (in LBG-8 mg InG/ml), thereby suggeting that these chemicals also inhibit fatty acid synthesis.

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Transcriptional and Mycolic Acid Profiling in Mycobacterium bovis BCG In Vitro Show an Effect for c-di-GMP and Overlap between Dormancy and Biofilms

  • Cruz, Miguel A. De la;Ares, Miguel A.;Rodriguez-Valverde, Diana;Vallejo-Cardona, Alba Adriana;Flores-Valdez, Mario Alberto;Nunez, Iris Denisse Cota;Aceves-Sanchez, Michel de Jesus;Lira-Chavez, Jonahtan;Rodriguez-Campos, Jacobo;Bravo-Madrigal, Jorge
    • Journal of Microbiology and Biotechnology
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    • 제30권6호
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    • pp.811-821
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    • 2020
  • Mycobacterium tuberculosis produces mycolic acids which are relevant for persistence, recalcitrance to antibiotics and defiance to host immunity. c-di-GMP is a second messenger involved in transition from planktonic cells to biofilms, whose levels are controlled by diguanylate cyclases (DGC) and phosphodiesterases (PDE). The transcriptional regulator dosR, is involved in response to low oxygen, a condition likely happening to a subset of cells within biofilms. Here, we found that in M. bovis BCG, expression of both BCG1416c and BCG1419c genes, which code for a DGC and a PDE, respectively, decreased in both stationary phase and during biofilm production. The kasA, kasB, and fas genes, which are involved in mycolic acid biosynthesis, were induced in biofilm cultures, as was dosR, therefore suggesting an inverse correlation in their expression compared with that of genes involved in c-di-GMP metabolism. The relative abundance within trehalose dimycolate (TDM) of α-mycolates decreased during biofilm maturation, with methoxy mycolates increasing over time, and keto species remaining practically stable. Moreover, addition of synthetic c-di-GMP to mid-log phase BCG cultures reduced methoxy mycolates, increased keto species and practically did not affect α-mycolates, showing a differential effect of c-di-GMP on keto- and methoxy-mycolic acid metabolism.