Journal of Comparative Physiology B

, Volume 164, Issue 7, pp 561–569 | Cite as

d-Alanine metabolism in the lucinid clam Lucinoma aequizonata

  • S. Wiley
  • H. Felbeck


The chemoautotrophic symbiont-bearing clam Lucinoma aequizonata contains very high levels of free d-alanine in all tissues. The possible sources for this amino acid and its involvement in the clams' metabolism were investigated. Very low levels of d-alanine (generally below 1 μmol·l-1) were measured in the sediment porewaters from the habitat of the clams. Experiments with 14C-labeled tracers demonstrate an active metabolism of d-alanine in the clams rather than a role as inert waste product. d-alanine is metabolized at about 0.12 μmol·g fw-1·h-1. Label from aspartate, but not glucose and CO2, is incorporated into d-alanine. Incubation with labeled d-alanine did not result in formation of radioactive l-alanine. Tests for alanine racemase (EC and d-amino acid oxidase (EC did not show activity in either gill, i.e. symbiont and host, or foot tissue. d-Alanine amino transferase (EC 2.6.1.b.) was demonstrated in gill and foot tissues. Two sources for d-alanine are proposed: a degradation of cell walls of symbiotic bacteria and production by the host using a d-specific alanine transaminase.

Key words

d-Alanine Chemoautotrophic symbiosis Lucinidae Clam, Lucinoma 



amino acid(s)


fresh weight


high-performance liquid chromatography


methyl benzethonium hydroxyde






tricarbonic acid


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Allen JA (1958) On the basic form and adaptations to the habitat in the Lucinaceae (Eulamellibranchia). Phil Trans R Soc B 241:421–484Google Scholar
  2. Aswad DD (1984) Determination of d-and l-aspartate in amino acid mixtures by high-performance liquid chromatography after derivatization with a chiral adduct of o-phthaldialdehyde. Anal Biochem 137:405–409Google Scholar
  3. Bada JL (1982) Racemization of amino acids in nature. Interdisciplinary Sci Rev 7:30–46Google Scholar
  4. Bada JL, Brown SE (1980) Amino acid racemization in living mammals-biochronological applications. Trends Biochem Sci 5: R3-R5Google Scholar
  5. Bada JL, Lee C (1977) Decomposition and alteration of organic compounds dissolved in sea water. Mar Chem 5:527–534Google Scholar
  6. Berg CP (1953) Physiology of the d-amino acids. Physiol Rev 33:145–189Google Scholar
  7. Berg CJ, Alatalo P (1984) Potential of chemosynthesis in molluscan mariculture. Aquaculture 39:165–179Google Scholar
  8. Cary SC, Vetter RD, Felbeck H (1989) Habitat characterization and nutritional strategies of the endosymbiont-bearing bivalve Lucinoma aequizonata. Mar Ecol Prog Ser 55:31–45Google Scholar
  9. Corrigan JJ (1969) d-Amino acids in animals. Science 164:142–149Google Scholar
  10. D'Aniello A, Giuditta A (1977) Identification of d-aspartic acid in the brain of Octopus vulgaris Lam. J Neurochem 29:1053–1057Google Scholar
  11. D'Aniello A, Giuditta A (1978) Presence of d-aspartate in squid axoplasm and in other regions of the cephalopod nervous system. J Neurochem 31:1107–1108Google Scholar
  12. D'Aniello A, Giuditta A (1980) Presence of d-alanine in crustacean muscle and hepatopancreas. Comp Biochem Physiol 66B:319–322Google Scholar
  13. Distel DL, Felbeck H (1987) Endosymbiosis in the lucinid clams Lucinoma aequizonata, Lucinoma annulata and Lucina floridana: a reexamination of the functional morphology of the gills as bacteria-bearing organs. Mar Biol 96:79–86Google Scholar
  14. Distel DL, Felbeck H (1988a) Pathways of inorganic carbon fixation in the endosymbiont-bearing lucinid clam Lucinoma aequizonata. Part 2. Analysis of the individual contributions of host and symbiont cells to inorganic carbon assimilation. J Exp Zool 247:11–22Google Scholar
  15. Distel DL, Felbeck H (1988b) Pathways of inorganic carbon fixation in the endosymbiont-bearing lucinid clam Lucinoma aequizonata. Part 1. Purification and characterization of the endosymbiotic bacteria. J Exp Zool 247:1–10Google Scholar
  16. Felbeck H (1980) Investigations on the role of the amino acids in anaerobic metabolism of the lugworm Arenicola marina L. J Comp Physiol 137:183–192Google Scholar
  17. Felbeck H (1985) Occurrence and metabolism of d-aspartate in the gutless bivalve Solemya reidi. J Exp Zool 234:145–149Google Scholar
  18. Felbeck H, Wiley S (1987) Free d-amino acids in the tissues of marine bivalves. Biol Bull 173:252–259Google Scholar
  19. Fiala-Medioni A, Metivier C, Herry A, Le Pennec M (1986) Ultrastructure of the gill of the hydrothermal-vent mytilid Bathymodiolus sp. Mar Biol 92:65–72Google Scholar
  20. Graßl M, Supp M (1983a) l-Alanine: determination with alanine aminotransferase and lactate dehydrogenase. In: Bermeyer HU (ed) Methods of enzymatic analysis. Verlag Chemie, Weinheim, pp 345–349Google Scholar
  21. Graßl M, Supp M (1983b) d-alanine. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Verlag Chemie, Weinheim, pp 336–340Google Scholar
  22. Hiatt AC, McIndoo J, Malmberg RL (1986) Regulation of polyamine biosynthesis in tobacco. Effects of inhibitors and exogenous polyamines on arginine decarboxylase, ornithine decarboxylase, and S-adenosylmethionine decarboxylase. J Biol Chem 261:1293–1298Google Scholar
  23. Jackson JBC (1973) The ecology of molluses of Thalassia communities, Jamaica, West Indies. Part 1. Distribution, environmental physiology, and ecology of common shallow-water species. Bull Mar Sci 23:313–350Google Scholar
  24. Katz E, Derrain AL (1977) Peptide antibiotics of Bacillus-chemistry, biogenesis, and possible functions. Bacteriol Rev 41:449–474Google Scholar
  25. Lee C, Bada JL (1977) Dissolved amino acids in the equatorial Pacific, the Sargasso Sea, and Biscayne Bay. Limnol Oceanogr 22:502–510Google Scholar
  26. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  27. Manahan DT, Wright SH, Stephens GC (1983) Simultaneous determination of net uptake of 16 amino acids by a marine bivalve. Am Physiol Soc 244:832–838Google Scholar
  28. Maniatis T, Fritsch EF, Sambrook J (1982) In: Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory, New York, p 68Google Scholar
  29. Martinez-Carrion M, Jenkins WT (1965) d-Alanine-d-glutamate transaminase. Part 1. Purification and characterization. J Biol Chem 240:3538–3546Google Scholar
  30. Matsushima O, Katayama H, Yamada K, Kado Y (1984) Occurrence of free d-alanine and alanine racemase activity in bivalve molluscs with special reference to intracellular osmoregulation. Mar Biol Lett 5:217–225Google Scholar
  31. Meister A (1965) Biochemistry of the amino acids, vol I, 2nd edn. Academic Press, New York, pp 113–118, 220–224, 297–304, 332–375Google Scholar
  32. Mor A, Delfour A, Nicolas P (1991) Identification of a d-alanine containing polypeptide precursor for the peptide opiod, dermorphin. J Biol Chem 266:6264–6270Google Scholar
  33. Muneoka Y, Kobayashi M (1992) Comparative aspects of structure and action of molluscan neuropeptides. Experientia 48:448–456Google Scholar
  34. Nagata Y, Akino T (1990) d-Amino acids in mouse tissues are not of microbial origin. Experientia 46:466–468Google Scholar
  35. Preston RL (1985) Distribution and transport of d-amino acids in marine invertebrates. Bull Mt Desert Isl Biol Lab 25:100–103Google Scholar
  36. Preston RL (1987a) Occurrence of d-amino acids in higher organisms: a survey of the distribution of d-amino acids in marine invertebrates. Comp Biochem Physiol 87:55–62Google Scholar
  37. Preston RL (1987b) d-Alanine transport and metabolism by the coelomocytes of the bloodworm, Glycera dibranchiata (Polychaeta). Comp Biochem Physiol 87:63–71Google Scholar
  38. Schöttler U, Wienhausen G, Zebe E (1983) The mode of energy production in the lugworm Arenicola marina at different oxygen concentrations. J Comp Physiol 149:547–555Google Scholar
  39. Schweimanns M, Felbeck H (1985) Significance of the occurrence of chemoautotrophic bacterial endosymbionts in lucinid clams from Bermuda. Mar Ecol Prog Ser 24:113–120Google Scholar
  40. Stein JL, Haygood M, Felbeck H (1989) Diversity of ribulose bisphosphate carboxylase genes in thiotrophic symbioses. In: Nardon P et al (eds) Endocytobiology IV. Institute National de la Recherche Agronomique, Paris, pp 343–348Google Scholar
  41. Vetter RD (1985) Elemental sulfur in the gills of three species of clams containing chemoautotrophic symbiotic bacteria: a possible inorganic energy storage compound. Mar Biol 88:33–42Google Scholar
  42. Virkar R, Webb KL (1970) Free amino acid composition of the soft-shell clam Mya arenaria in relation to salinity of the medium. Comp Biochem Physiol 32:775–783Google Scholar
  43. Zurburg W, De Zwaan A (1981) The role of amino acids in anaerobiosis and osmoregulation in bivalves. J Exp Zool 215:315–325Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • S. Wiley
    • 1
  • H. Felbeck
    • 1
  1. 1.Scripps Institution of Oceanography 0202University of California San DiegoLa JollaUSA

Personalised recommendations