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. 1990 Mar;172(3):1380–1384. doi: 10.1128/jb.172.3.1380-1384.1990

Enzymes of ammonia assimilation in hyphae and vesicles of Frankia sp. strain CpI1.

N A Schultz 1, D R Benson 1
PMCID: PMC208609  PMID: 1968454

Abstract

Frankia spp. are filamentous actinomycetes that fix N2 in culture and in actinorhizal root nodules. In combined nitrogen-depleted aerobic environments, nitrogenase is restricted to thick-walled spherical structures, Frankia vesicles, that are formed on short stalks along the vegetative hyphae. The activities of the NH4(+)-assimilating enzymes (glutamine synthetase [GS], glutamate synthase, glutamate dehydrogenase, and alanine dehydrogenase) were determined in cells grown on NH4+ and N2 and in vesicles and hyphae from N2-fixing cultures separated on sucrose gradients. The two frankial GSs, GSI and GSII, were present in vesicles at levels similar to those detected in vegetative hyphae from N2-fixing cultures as shown by enzyme assay and two-dimensional polyacrylamide gel electrophoresis. Glutamate synthase, glutamate dehydrogenase, and alanine dehydrogenase activities were restricted to the vegetative hyphae. Vesicles apparently lack a complete pathway for assimilating ammonia beyond the glutamine stage.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aharonowitz Y., Friedrich C. G. Alanine dehydrogenase of the beta-lactam antibiotic producer Streptomyces clavuligerus. Arch Microbiol. 1980 Mar;125(1-2):137–142. doi: 10.1007/BF00403210. [DOI] [PubMed] [Google Scholar]
  2. Bender R. A., Janssen K. A., Resnick A. D., Blumenberg M., Foor F., Magasanik B. Biochemical parameters of glutamine synthetase from Klebsiella aerogenes. J Bacteriol. 1977 Feb;129(2):1001–1009. doi: 10.1128/jb.129.2.1001-1009.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bergersen J. F., Turner G. L. Nitrogen fixation by the bacteroid fraction of breis of soybean root nodules. Biochim Biophys Acta. 1967 Aug 29;141(3):507–515. doi: 10.1016/0304-4165(67)90179-1. [DOI] [PubMed] [Google Scholar]
  4. Boland M. J., Benny A. G. Enzymes of nitrogen metabolism in legume nodules. Purification and properties of NADH-dependent glutamate synthase from lupin nodules. Eur J Biochem. 1977 Oct 3;79(2):355–362. doi: 10.1111/j.1432-1033.1977.tb11816.x. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Callaham D., Deltredici P., Torrey J. G. Isolation and Cultivation in vitro of the Actinomycete Causing Root Nodulation in Comptonia. Science. 1978 Feb 24;199(4331):899–902. doi: 10.1126/science.199.4331.899. [DOI] [PubMed] [Google Scholar]
  7. Carlson T. A., Martin G. B., Chelm B. K. Differential transcription of the two glutamine synthetase genes of Bradyrhizobium japonicum. J Bacteriol. 1987 Dec;169(12):5861–5866. doi: 10.1128/jb.169.12.5861-5866.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Darrow R. A., Knotts R. R. Two forms of glutamine synthetase in free-living root-nodule bacteria. Biochem Biophys Res Commun. 1977 Sep 23;78(2):554–559. doi: 10.1016/0006-291x(77)90214-5. [DOI] [PubMed] [Google Scholar]
  9. Edmands J., Noridge N. A., Benson D. R. The actinorhizal root-nodule symbiont Frankia sp. strain CpI1 has two glutamine synthetases. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6126–6130. doi: 10.1073/pnas.84.17.6126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fuchs R. L., Keister D. L. Comparative properties of glutamine synthetases I and II in Rhizobium and Agrobacterium spp. J Bacteriol. 1980 Nov;144(2):641–648. doi: 10.1128/jb.144.2.641-648.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Johansson B. C., Gest H. Inorganic nitrogen assimilation by the photosynthetic bacterium Rhodopseudomonas capsulata. J Bacteriol. 1976 Nov;128(2):683–688. doi: 10.1128/jb.128.2.683-688.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kanamori K., Weiss R. L., Roberts J. D. Ammonia assimilation in Bacillus polymyxa. 15N NMR and enzymatic studies. J Biol Chem. 1987 Aug 15;262(23):11038–11045. [PubMed] [Google Scholar]
  13. Kanamori K., Weiss R. L., Roberts J. D. Role of glutamate dehydrogenase in ammonia assimilation in nitrogen-fixing Bacillus macerans. J Bacteriol. 1987 Oct;169(10):4692–4695. doi: 10.1128/jb.169.10.4692-4695.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kenealy W. R., Thompson T. E., Schubert K. R., Zeikus J. G. Ammonia assimilation and synthesis of alanine, aspartate, and glutamate in Methanosarcina barkeri and Methanobacterium thermoautotrophicum. J Bacteriol. 1982 Jun;150(3):1357–1365. doi: 10.1128/jb.150.3.1357-1365.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mazzucco C. E., Benson D. R. [14C]methylammonium transport by Frankia sp. strain CpI1. J Bacteriol. 1984 Nov;160(2):636–641. doi: 10.1128/jb.160.2.636-641.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Meeks J. C., Wolk C. P., Lockau W., Schilling N., Shaffer P. W., Chien W. S. Pathways of assimilation of [13N]N2 and 13NH4+ by cyanobacteria with and without heterocysts. J Bacteriol. 1978 Apr;134(1):125–130. doi: 10.1128/jb.134.1.125-130.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
  18. Murry M. A., Fontaine M. S., Tjepkema J. D. Oxygen protection of nitrogenase in Frankia sp. HFPArI3. Arch Microbiol. 1984 Oct;139(2-3):162–166. doi: 10.1007/BF00401993. [DOI] [PubMed] [Google Scholar]
  19. Murry M. A., Zhongze Z., Torrey J. G. Effect of O2 on vesicle formation, acetylene reduction, and O2-uptake kinetics in Frankia sp. HFPCcI3 isolated from Casuarina cunninghamiana. Can J Microbiol. 1985 Sep;31(9):804–809. doi: 10.1139/m85-151. [DOI] [PubMed] [Google Scholar]
  20. Noridge N. A., Benson D. R. Isolation and nitrogen-fixing activity of Frankia sp. strain CpI1 vesicles. J Bacteriol. 1986 Apr;166(1):301–305. doi: 10.1128/jb.166.1.301-305.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  22. Orr J., Haselkorn R. Regulation of glutamine synthetase activity and synthesis in free-living and symbiotic Anabaena spp. J Bacteriol. 1982 Nov;152(2):626–635. doi: 10.1128/jb.152.2.626-635.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Parsons R., Silvester W. B., Harris S., Gruijters W. T., Bullivant S. Frankia vesicles provide inducible and absolute oxygen protection for nitrogenase. Plant Physiol. 1987 Apr;83(4):728–731. doi: 10.1104/pp.83.4.728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schultz N. A., Benson D. R. Developmental potential of Frankia vesicles. J Bacteriol. 1989 Dec;171(12):6873–6877. doi: 10.1128/jb.171.12.6873-6877.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Thomas J., Meeks J. C., Wolk C. P., Shaffer P. W., Austin S. M. Formation of glutamine from [13n]ammonia, [13n]dinitrogen, and [14C]glutamate by heterocysts isolated from Anabaena cylindrica. J Bacteriol. 1977 Mar;129(3):1545–1555. doi: 10.1128/jb.129.3.1545-1555.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tisa L. S., Ensign J. C. Isolation and nitrogenase activity of vesicles from Frankia sp. strain EAN1pec. J Bacteriol. 1987 Nov;169(11):5054–5059. doi: 10.1128/jb.169.11.5054-5059.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tunlid A., Schultz N. A., Benson D. R., Steele D. B., White D. C. Differences in fatty acid composition between vegetative cells and N(2)-fixing vesicles of Frankia sp. strain CpI1. Proc Natl Acad Sci U S A. 1989 May;86(9):3399–3403. doi: 10.1073/pnas.86.9.3399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tyler B. Regulation of the assimilation of nitrogen compounds. Annu Rev Biochem. 1978;47:1127–1162. doi: 10.1146/annurev.bi.47.070178.005403. [DOI] [PubMed] [Google Scholar]

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