Amino acid utilization patterns in clostridial taxonomy
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The polyamide layer technique for the chromatographic separation of dimethylaminonaphthalene sulphonyl amino acids has been adapted to the qualitative analysis of amino acids in media before and after the growth of micro-organisms. The method has been used to study the amino acids metabolized by cultures of proteolytic clostridia growing in a medium consisting of an acid hydrolysate of casein as a source of amino acids and small amounts of yeast extract and trypticase as sources of growth factors. The chromatograms of the media after growth showed which amino acids were used and which new amino acids were produced.
Clostridium botulinum type F (proteolytic), C. ghoni, C. mangenoti and C. putrificum were found to reduce proline to 5-aminovaleric acid and to produce 2-aminobutyric acid, properties they shared with C. sporogenes and C. sticklandii. C. botulinum type G and C. subterminale used glycine, lysine, serine, and arginine but in contrast to C. sticklandii they neither reduced proline to 5-aminovaleric acid nor produced 2-aminobutyric acid. Both organisms oxidized phenylalanine, tyrosine and tryptophan to phenylacetic acid, p-hydroxyphenyl acetic acid and indole acetic acid respectively. C. lituseburense and C. scatologenes used serine, threonine and arginine and produced 2-amino butyric acid and ornithine. C. lentoputrescens, C. limosum and C. malenomenatum resembled C. tetanomorphum by using glutamic acid and tyrosine. The chromatograms always showed the physiological group to which an organism belonged and in some cases were characteristic of the species.
Key words2-aminobutyric acid 5-aminovaleric acid Glutamic acid Lysine Proline Tyrosine Polyamide layer Chromatography Clostridia
the sulphonyl chloride
the sulphonic acid
steam volatile fatty acid
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- Buckel, W., Barker, H. A.: Two pathways of glutamate by anaerobic bacteria. J. Bacteriol. 117, 1248–1260 (1974)Google Scholar
- Ciccarelli, A. S., Whaley, D. N., McCroskey, L. M., Giminez, D. F., Dowell, V. R. Jr., Hatheway, C. L.: Cultural and physiological characteristics of Clostridium botulinum type G and the susceptibility of certain animals to its toxin. Appl. Env. Microbiol. 34, 843–848 (1977)Google Scholar
- Elsden, S. R., Hilton, M. G.: Volatile acid production from threonine, valine, leucine and isoleucine by clostidia. Arch. Microbiol. 117, 165–172 (1978)Google Scholar
- Elsden, S. R., Hilton, M. G., Waller, J. M.: The end products of the metabolism of aromatic amino acids by clostridia. Arch. Microbiol. 107, 283–288 (1976)Google Scholar
- Guillaume, J., Beerens, H., Osteux, H.: Production de gaz carbonique et fermentation du glycocolle par Clostridium histolyticum. Ann. Inst. Pasteur 91, 721–726 (1956)Google Scholar
- Lee, M.-L., Safille, A.: Improved solvent for thin-layer chromatography of DNS-amino acids. J. Chromatogr. 116, 462–464 (1976)Google Scholar
- Mead, G. C.: The amino acid-fermenting clostridia. J. Gen. Microbiol. 67, 47–56 (1971)Google Scholar
- Smith, L. D. S., Hobbs, G.: Genus III Clostridium. In: Bergey's manual of determinative bacteriology 8th ed. (R. E. Buchanan, N. E. Gibbons, eds.), pp. 551–572. Baltimore: Williams and Wilkins 1974Google Scholar
- Stadtman, T. C.: On the metabolism of an amino acid fermenting Clostridium. J. Bacteriol. 67, 314–319 (1954)Google Scholar
- Stickland, L. H.: Studies on the metabolism of the strict anaerobes (genus Clostridium). I. The chemical reactions by which Cl. sporogenes obtains its energy. Biochem. J. 28, 1746–1759 (1934)Google Scholar
- Stickland, L. H.: Studies in the metabolism of the strict anaerobes (genus Clostridium). II. The reduction of proline by Cl. sporogenes. Biochem. J. 29, 288–290 (1935)Google Scholar
- Woods, D. D., Clifton, C. E.: Studies in the metabolism of the strict anaerobes (genus Clostridium). VI. Hydrogen production and amino acid utilization by Clostridium tetanomorphum. Biochem. J. 31, 1774–1788 (1937)Google Scholar
- Woods, K. R., Wang, K.-T.: Separation of dansyl-amino acids by polyamide layer chromatography. Biochim. Biophys. Acta 133, 369–370 (1967)Google Scholar