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.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
the sulphonyl chloride
- DNS-NH2 :
the sulphonic acid
steam volatile fatty acid
Buckel, W., Barker, H. A.: Two pathways of glutamate by anaerobic bacteria. J. Bacteriol. 117, 1248–1260 (1974)
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)
Elsden, S. R., Hilton, M. G.: Volatile acid production from threonine, valine, leucine and isoleucine by clostidia. Arch. Microbiol. 117, 165–172 (1978)
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)
Guillaume, J., Beerens, H., Osteux, H.: Production de gaz carbonique et fermentation du glycocolle par Clostridium histolyticum. Ann. Inst. Pasteur 91, 721–726 (1956)
Lee, M.-L., Safille, A.: Improved solvent for thin-layer chromatography of DNS-amino acids. J. Chromatogr. 116, 462–464 (1976)
Mead, G. C.: The amino acid-fermenting clostridia. J. Gen. Microbiol. 67, 47–56 (1971)
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 1974
Stadtman, T. C.: On the metabolism of an amino acid fermenting Clostridium. J. Bacteriol. 67, 314–319 (1954)
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)
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)
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)
Woods, K. R., Wang, K.-T.: Separation of dansyl-amino acids by polyamide layer chromatography. Biochim. Biophys. Acta 133, 369–370 (1967)
About this article
Cite this article
Elsden, S.R., Hilton, M.G. Amino acid utilization patterns in clostridial taxonomy. Arch. Microbiol. 123, 137–141 (1979). https://doi.org/10.1007/BF00446812
- 2-aminobutyric acid
- 5-aminovaleric acid
- Glutamic acid
- Polyamide layer