Abstract
The anaerobic fungusPiromyces sp. strain E2 appeared restricted in nitrogen utilization. Growth was only supported by ammonium as source of nitrogen. Glutamine also resulted in growth, but this was due to release of ammonia rather than to uptake and utilization of the amino acid. The fungus was not able to grow on other amino acids, albumin, urea, allantoin, or nitrate. Assimilation of ammonium is very likely to be mediated by NADP-linked glutamate dehydrogenase (NADP-GDH) and glutamine synthetase (GS). One transaminating activity, glutamate-oxaloacetate transaminase (GOT), was demonstrated. Glutamate synthase (GOGAT), NAD-dependent glutamate dehydrogenase (NAD-GDH), and the transaminating activity glutamate-pyruvate transaminase (GPT) were not detected in cell-free extracts ofPiromyces sp. strain E2. Specific enzyme activities of both NADP-GDH and GS increased four-to sixfold under nitrogen-limiting conditions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- GDH :
-
Glutamate dehydrogenase
- GOGAT :
-
Glutamate synthase
- GOT :
-
Glutamate-oxaloacetate transaminase
- GPT :
-
Glutamate-pyruvate transaminase
- GS :
-
Glutamine synthetase
References
Arst HM, Parbtani AAM, Cove DJ (1975) A mutant ofAspergillus nidulans defective in NAD-linked glutamate dehydrogenase. Mol Gen Genet 138:165–171
Asao N, Ushida K, Kojima Y (1993) Proteolytic activity of rumen fungi belonging to the generaNeocallimastix andPiromyces. Lett Appl Microbiol 16:247–250
Baars JJP, Op den Camp HJM, Hermans JHM Mike V, Van der Drift C, Van Griensven LJLD, Vogels GD (1994) Nitrogen assimilating enzymes in the white button mushroomAgaricus bisporus. Microbiology 140:1161–1168
Baars JJP, Op den Camp HJM, Van der Drift C, Van Griensven LJLD, Vogels GD (1995a) Regulation of nitrogen metabolising enzymes in the commercial mushroomAgaricus bisporus. Curr Microbiol 31:345–350
Baars JJP, Op den Camp HJM, Van Hoek AHAM, Van der Drift C, Van Griensven LJLD, Vogels GD (1995b) Purification and characterization of NADP-dependent glutamate dehydrogenase from the commercial mushroomAgaricus bisporus. Curr Microbiol 30:211–217
Bergmeyer HU, Beutler HO (1985) Determination of ammonia. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 8. Verlag Chemie, Weinheim Basel, pp 454–461
Brown CM, Macdonald-Brown DS, Meers JL (1974) Physiological aspects of microbial inorganic nitrogen metabolism. Adv Microb Physiol 11:1–52
Burn VJ, Turner PR, Brown CM (1974) Aspects of inorganic nitrogen assimilation in yeasts. Antonie van Leeuwenhoek 40:93–102
genetet I, Martin F, Stewart GR (1984) Nitrogen assimilation in mycorrhizas. Plant Physiol 76:395-39
Goldin BR, Frieden C (1971)l-Glutamate dehydrogenases. Curr Top Cell Regul 4:77–117
Gulati SK, Ashes JR, Gordon GLR, Connel PJ, Rogers PL (1989) Nutritional availability of amino acids from the rumen anaerobic fungusNeocallimastix sp. LM1 in sheep. J Agric Sci (Camb) 113:383–387
Holmes AR, Collings A, Famden KJF, Shepherd MG (1989) Ammonium assimilation byCandida albicans and other yeasts: evidence for activity of glutamate synthase. J Gen Microbiol 135:1423–1430
Kusnan MB, Berger MG, Fock HP (1987) The involvement of glutamine synthetase/glutamate synthase in ammonia assimilation byAspergillus nidulans. J Gen Microbiol 133:1235–1242
Lara M, Blanco L, Campomanes M, calva E, Palacios R, Mora J (1982) Physiology of ammonium assimilation inNeurospora crassa. J Bacteriol 150:105–112
Lejohn HB (1971) Enzyme regulation, lysine pathways and cell wall structures as indicators of major lines of evolution of fungi. Nature 231:164–168
Marounek M, Vovk SJ (1992) Distribution of radioactivity of14C-amino acids added to the medium in cells and metabolites in cultures of rumen fungi. Reprod Nutr Dev 32:129–133
Marvin-Sikkema FD, Lahpor GA, Kraak MN, Gottschal JC, Prins RA (1992) Characterization of an anaerobic fungus from Ilama faeces. J Gen Microbiol 138:2235–2241
Meers JL, Tempest DW, Brown CM (1970) Glutamine(amide):2-oxoglutarate amino transferase oxido-reductase (NADP), an enzyme involved in the synthesis of glutamate by some bacteria. J Gen Microbiol 64:187–194
Michel V, Fonty G, Millet L, Bonnemoy F, Gouet P (1993) In vitro study of the proteolytic activity of rumen anaerobic fungi. FEMS Microbiol Lett 110:5–10
Miller SM, Magasanik B (1990) Role of NAD-linked glutamate dehydrogenase in nitrogen metabolism inSaccharomyces cerevisiae. J Bacteriol 172:4927–4935
Orpin CG (1975) Studies on the rumen flagellateNeocallimastix frontalis. J Gen Microbiol 91:249–262
Orpin CG (1988) Nutrition and biochemistry of anaerobic Chitridiomycetes. Biosystems 21:365–370
Orpin CG, Greenwood Y (1986) Nutritional and gemination requirements of the rumen chytridiomyceteNeocallimastix patriciarum. Trans Br Mycol Soc 86:103–109
Pateman JA (1969) Regulation of synthesis of glutamate dehydrogenase and glutamine synthetase in micro-organisms. Biochem J 115:769–775
Sakurada M, Morgavi DP, Tomita Y, Onodera R (1994) Ureolytic activity of anaerobic rumen fungi,Piomyces sp. OTS3 andNeocallimastix sp. OTS4. An Sci Tech 65:950–955
Sanwal BD, Lata M (1961) The occurrence of two different glutamic dehydrogenases inNeurospora. Can J Microbiol 7:319–328
Sanwal BD, Lata M (1962) Concurrent regulation of glutamic acid dehydrogenases ofNeurospora. Arch Biochem Biophys 97:582–588
Schwartz T, Kusnan MB, Fock HP (1991) The invovement of glutamate dehydrogenase and glutamine synthetase/glutamate synthase in ammonia assimilation by the basidiomycete fungusStropharia semiglobata. J Gen Microbiol 137:2253–2258
Sleat R, Mah RA (1984) Quantitative method for colorimetric determination of formate in fermentation media. Appl Environ Microbiol 47:884–885
Stewart GR, Mann AF, Fentem PA (1980) Enzymes of glutamate formation: glutamate dehydrogenase, glutamine synthetase and glutamate synthase. In Miflin BJ (ed) The biochemistry of plants, a comprehensive treatise, vol 5. Academic Press, New York, pp 271–327
Teunissen MJ, Marras SAE, Op den Camp HJM, Vogels GD (1989) An improved method for the quantification of alcohols, volatile fatty acids, and lactate or 2,3-butanediol in biological samples. J Microbiol Methods 10:247–254
Teunissen MJ, Op den Camp HJM, Huis in 't Veld JHJ, Vogels GD (1991a) Comparison of growth characteristics of anaerobic fungi from ruminant and non-ruminant herbivores during cultivation in a defined medium. Gen Microbiol 137:1401–1408
Teunissen MJ, Smits TAM, Op den Camp HJM, Huis in 't Veld JHJ, Vogels GD (1991b) Fermentation of cellulose and production of cellulolytic and xylanolytic enzymes by anaerobic fungi from ruminant and non-ruminant herbivores. Arch Microbiol 156:290–296
Trinci APJ, Davies DR, Gull K, Lawrence MI, Nielsen BB, Rickers A, Theodorou MK (1994) Anaerobic fungi in herbivorous animals. Mycol Res 98:129–152
Wallace RJ, Cotta MA (1988) Metabolism of nitrogen-containing compounds. In: Hobson PN (ed) The rumen microbial ecosystem. Elsevier Applied Science, London, pp 217–249
Wallace RJ, Joblin KN (1985) Proteolytic activity of a rumen anaerobic fungus. FEMS Microbiol Lett 29:19–25
Wallace RJ, Munro CA (1986) Influence of the rumen anaerobic fungusNeocallimastix frontalis on the proteolytic activity of a defined mixture of rumen bacteria growing on a solid substrate. Lett Appl Microbiol 3:23–26
Wooton JF, Argenzio RA (1975) Nitrogen utilization within equine large intestine. Am J Physiol 229:1061–1067
Younes H, Garleb K, Behr S, Rémésy C, Demigné C (1994) Fermentable fibers or oligosaccharides reduce urinary nitrogen excretion by increasing urea disposal in the rat cecum. J Nutr 125:1010–1016
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Dijkerman, R., Ledeboer, J., Verhappen, A.B.M. et al. The anaerobic fungusPiromyces sp. strain E2: nitrogen requirement and enzymes involved in primary nitrogen metabolism. Arch. Microbiol. 166, 399–404 (1996). https://doi.org/10.1007/BF01682986
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01682986