Influence of Amino Acids on Growth and Cell Wall Composition of Methanobacteriales

König, Helmut

doi:10.1099/00221287-131-12-3271

Microbiology

1985

DOI: 10.1099/00221287-131-12-3271

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Influence of Amino Acids on Growth and Cell Wall Composition of Methanobacteriales

Helmut König

Abstract: 1Methanogenic bacteria with pseudomurein sacculi incubated with elevated concentrations of amino acids showed growth inhibition, changes in morphology and, under certain conditions, lysis. Methanobacterium therrnoautotrophicum incorporated the amino acids glycine, threonine, ornithine and, in small amounts, aspartic acid into the sacculi, but not D-alanine, mesodiaminopimelic acid or the amino sugar galactosamine.

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1989

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Cited by 2 publications

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“…Formate does not appear to be a free intermediate for incorporation into either carbon of the acetyl group (11,25,26). Determination of a biosynthetic role for formate in M. thermoautotrophicum by substitution with other compounds may be difficult, since this microorganism probably lacks appropriate uptake mechanisms (7,12), growth can be inhibited by relatively high (millimolar) concentrations of organic compounds in media (12,13), and more than one compound may be required.…”

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Formate auxotroph of Methanobacterium thermoautotrophicum Marburg

1989

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A formate-requiring auxotroph of Methanobacterium thermoautotrophicum Marburg was isolated after hydroxylamine mutagenesis and bacitracin selection. The requirement for formate is unique and specific; combined pools of other volatile fatty acids, amino acids, vitamins, and nitrogen bases did not substitute for formate. Compared with those of the wild type, cell extracts of the formate auxotroph were deficient in formate dehydrogenase activity, but cells of all of the strains examined catalyzed a formate-carbon dioxide exchange activity. All of the strains examined took up a small amount (200 to 260 ,umol/liter) of formate (3 mM) added to medium. The results of the study of this novel auxotroph indicate a role for formate in biosynthetic reactions in this methanogen. Moreover, because methanogenesis from H2-C02 is not impaired in the mutant, free formate is not an intermediate in the reduction of CO2 to CH4.The autotrophic, thermophilic archaebacterium Methanobacterium thermoautotrophicum has several attractive features for the development of a genetic system in a methanogen, including well-characterized mutants (19,21), the presence of a cryptic plasmid (17) which may be suitable for the development of a plasmid vector (18), and a primitive natural transformation system for genetic exchange (30). While much of the research into developing a genetic exchange system in M. thermoautotrophicum in our laboratory has focused on mutants resistant to nitrogen base analogs (19, 28, 30; V. E. Worrell and D. P. Nagle, Jr., Abstr. Annu. Meet. Am. Soc. Microbiol. 1989, I22, p. 221; D. P. Nagle, Jr., Dev. Ind. Microbiol., in press), mutants with auxotrophic markers would also be useful for these investigations into the genetics of M. thermoautotrophicum. In this report we describe the isolation and partial characterization of a unique auxotroph, a formate-requiring strain of M. thermoautotrophicum.(A portion of this work has been presented elsewhere [R. S. Tanner and D. P. Nagle, Jr., Abstr. Annu. Meet. Am. Soc. Microbiol. 1988, I10, p. 182].) MATERIALS AND METHODS Strains and media. Strains of M. thermoautotrophicum used in this study were AH (ATCC 29096), obtained from R. S. Wolfe; Marburg (DSM 2133), obtained from H. Hippe; Kr, a kanamycin-resistant derivative of Marburg; and RT-103, a formate-requiring derivative of Kr. M. thermoautotrophicum Kr was isolated after prolonged incubation of a culture in medium containing 500 ,ug of kanamycin per ml. The MICs of kanamycin sulfate in basal medium for strains Marburg and Kr were 175 and 1,000 ,ug/ml, respectively.The basal medium contained the following, in grams per liter: NaCl, 0.8; NH4Cl, 1.0; KCI, 0.1; KH2PO4, 0.1; MgSO4. 7H20, 0.2; CaCl2 -2H20, 0.02; NaHCO3, 5.0; resazurin, 0.0005; cysteine hydrochloride, 0.4; and Na2S. 9H20, 0.4, as well as 10 ml of trace metal solution. The trace metal solution was adapted from that of Wolin et al. (29) and contained the following, in grams per liter: nitrilotriacetic acid, 2.0 (pH adjusted to 6 with KOH); MnSO4-H20, 1.0; Fe(NH4)2(SO4...

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Formate auxotroph of Methanobacterium thermoautotrophicum Marburg

1989

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Estimation of genome sizes of hyperthermophiles

et al. 1998

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Genomes of various hyperthermophilic and extremely thermophilic prokaryotes were analyzed with respect to size, physical organization, and 16S rDNA copy number. Our results show that all the genomes are circular, and they are in the size range of 1.6-1.8 Mb for Pyrodictium abyssi, Methanococcus igneus, Pyrobaculum aerophilum, Archaeoglobus fulgidus, Archaeoglobus lithotrophicus, and Archaeoglobus profundus (the two bacteria Fervidobacterium islandicum and Thermosipho africanus possess genomes of 1.5-Mb size). A systematic study of all validly described species of the order Sulfolobales revealed the existence of two classes of genome size for these archaea, correlating with phylogenetic analyses. The Metallosphaera-Acidianus group, plus Sulfolobus metallicus, have genomes of ca. 1.9 Mb; the other members of the order Sulfolobales group possess genome > 2.7 Mb. The special case of stygiolobus azoricus is discussed.

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Influence of Amino Acids on Growth and Cell Wall Composition of Methanobacteriales

Cited by 2 publications

References 19 publications

Formate auxotroph of Methanobacterium thermoautotrophicum Marburg

Formate auxotroph of Methanobacterium thermoautotrophicum Marburg

Estimation of genome sizes of hyperthermophiles

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