Abstract
The ability of Methanosarcina thermophila strain TM-1 to store a reserve polysaccharide was studied using both biochemical methods and thin-section electron microscopy. When grown under conditions of excess carbon and energy (either methanol or acetate) and limiting nitrogen, M. thermophila accumulated a polysaccharide which could be hydrolyzed to glucose by the enzyme amyloglucosidase. This polysaccharide reached levels of 20 mg polysaccharide per g protein in nitrogen-limited cells, while cells limited for carbon, as well as cells in the exponential phase of growth, did not accumulate significant amounts of this polysaccharide. Thin-section electron micrographs of M. thermophila showed glycogen-like inclusion granules in nitrogen-limited cells but not in carbon-limited or exponential-phase cells. These granules were stained by a polysaccharide-specific staining procedure, the PATO stain. The polysaccharide was purified from cell extracts, the iodine-polysaccharide complex gave a maximum absorption at between 500 and 510 nm. The polysaccharide was mobilized within 21 h by cells starved for a carbon/energy source. N-Limited (polysaccharide-containing) acetategrown cells could shift to methanogenesis from methanol more quickly than did C-limited acetate-grown cells lacking polysaccharide, and ATP levels remained higher in N-limited cells. The results are consistant with the hypothesis that this polysaccharide can provide carbon and energy for metabolic shifts but other storage compounds, such as polyphosphate, may also play a similar role.
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References
Beudeker RF, Kerver MWM, Keunen JG (1981) Occurrence, structure and function of intracellular polyglucose in the obligate chemolithotroph Thiobacillus neapolitanus. Arch Microbiol 129:221–226
Blaut M, Gottschalk G (1984) Coupling of ATP synthesis and methane formation from methanol and molecular hydrogen in Methanosarcina barkeri. Eur J Biochem 141:217–222
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of dye binding. Anal Biochem 72:248–254
Brana AF, Manzanal MB, Hardison C (1982) Characterization of intracellular polysaccharides of Streptomyces. Can J Microbiol 28:1320–1323
Cheng KJ, Brown RG, Costerton JW (1977) Characterization of a cytoplasmic plasmic reserve glucan from Ruminococcus albus. Appl Environ Microbiol 38:718–724
Evans JNS, Tolman CJ, Kanodia S, Roberts MF (1985) 2,3-Cyclopyrophosphoglycerate in methanogens. Evidence by 13C-NMR spectroscopy for a role in carbohydrate metabolism. Biochem 24:5693–5698
Fuchs G, Winter H, Steiner I, Stupperich E (1983) Enzymes of gluconeogenesis in the autotroph Methanobacterium thermoautotrophicum. Arch Microbiol 136:160–162
Gottschal JC, Pol A, Kuenen JG (1981) Metabolic flexibility of Thiobacillus A2 during substrate transitions in the chemostat. Arch Microbiol 129:23–28
Gunnarsson LAH, Ronnow PH (1982) Variation of the ATP-pool in thermophilic methanogenic bacteria during nitrogen or sulfur starvation. FEMS Microbiol Lett 14:317–320
Hardie LP, Balkwill DL, Stevens SE (1983) Effects of iron starvation on the ultrastructure of the cyanobacterium Agmenellum quadruplicatum. Appl Environ Microbiol 45:1007–1017
Hippe H, Caspari D, Fiebig K, Gottschalk G (1979) Utilization of trimethylamines and other N-methyl compounds for growth and methane formation by Methanosarcina barkeri. Proc Natl Acad Sci USA 76:494–498
Kamio Y, Terawaki Y, Nakajima T, Matsuda K (1981) Structure of glycogen produced by Selenomonas ruminantium. Agric Biol Chem 45:209–216
Kellenberger E, Ryter A, Sechaud J (1958) Electron microscope study of DNA plasms. J Biophys Biochem Cytol 4:671–678
König H, Stetter KO (1982) Isolation and characterization of Methanolobus tindarius, sp. nov., a coccoid methanogen growing only on methanol and methylamines. Zbl Bakt Hyg, Abt Orig C 3:478–490
König H, Skorko R, Zillig W, Reiter WD (1982) Glycogen in thermoacidophilic archaebacteria of the genera Sulfolobus, Thermoproteus, Desulfurococcus and Thermococcus. Arch Microbiol 132:297–303
König H, Nusser E, Stetter KO (1985) Glycogen in Methanolobus and Methanococcus. FEMS Microbiol Lett 28:265–269
Kreisl P, Kandler OC (1986) Chemical structure of the cell wall polymer of Methanosarcina. System Appl Microbiol 7:293–299
Lindner JGEM, Marcelis JH, de Vos NM, Hoog Kap-Korstanje (1979) Intracellular polysaccharide of Bacteroides fragilis. J Gen Microbiol 111:93–99
Linton JD, Cripps RE (1978) The occurrence and identification of intracellular polyglucose storage granules in Methylococcus NCIB 11083 grown in chemostate culture on methane. Arch Microbiol 117:41–48
Mah RA, Smith MR, Baresi L (1978) Studies on an acetatefermenting strain of Methanosarcina. Appl Environ Microbiol 35:1174–1184
Mah RA, Smith MR, Ferguson T, Zinder SH (1981) Methanogenesis from H2−CO2, methanol and acetate by Methanosarcina. In: Dalton H (ed) Microbial growth on C1 compounds. Heyden, London, pp 131–143
Murray PA, Zinder SH (1985) Nutritional requirements of Methanosarcina sp. strain TM-1. Appl Environ Microbiol 50:49–55
Nelson DR, Zeikus JG (1974) Rapid method for the radioisotopic analysis of gaseous end products of anaerobic metabolism. Appl Microbiol 28:258–261
Preiss J (1984) Bacterial glycogen synthesis and its regulation. Ann Rev Microbiol 38:418–458
Scherer PA, Bochem HP (1983) Ultrastructural investigation of 12 methanosarcinae and related species grown on methanol for occurrence of polyphosphate-like inclusions. Can J Microbiol 29:1190–1199
Seely RJ, Fahrney DE (1983) A novel diphospho-P, P′-diester from Methanobacterium thermoautotrophicum. J Biol Chem 258: 10835–10838
Seely RJ, Fahrney DE (1984) Levels of cyclic-2,3-diphosphoglycerate in Methanobacterium thermoautotrophicum during phosphate limitation. J Bacteriol 160:50–54
Sirevag R (1975) Photoassimilation of acetate and metabolism of carbohydrate in Chlorobium thiosulfatophilum. Arch Microbiol 104:105–111
Smith MR, Mah RA (1978) Growth and methanogenesis by Methanosarcina strain 227 on acetate and methanol. Appl Environ Microbiol 36:870–879
Smith MR, Zinder SH, Mah RA (1980) Microbial methanogenesis from acetate. Process Biochem 15:34–39
Sowers KR, Johnson JL, Ferry JG (1984) Phylogenetic relationships among the methylotrophic methane-producing bacteria and emendation of the family Methanosarcinaceae. Int J Syst Bacteriol 34:444–450
Tolman CJ, Kanodia S, Roberts MF, Daniels L (1986) 31P-NMR spectra of methanogens: 2,3-cyclopyrophospuglycerate is detectable only in methanobacteria strains. Biochem Biophys Acta 886:345–352
Venable JH, Coggeshall R (1965) A simplified lead citrate stain for use in electron microscopy. J Cell Biol 25:407–408
Zeikus JG (1977) The biology of methanogenic bacteria. Bacterial Rev 41:514–541
Zhilina TN (1976) Biotypes of Methanosarcina. Mikrobiol 45:481–489
Zinder SH, Elias AF (1985) Growth substrate effects on acetate and methanol catabolism in Methanosarcina sp. strain TM-1. J Bacteriol 163:317–323
Zinder SH, Mah RA (1979) Isolation and characterization of a thermophilic strain of Methanosarcina unable to use H2−CO2 for methanogenesis. Appl Environ Microbiol 38:996–1008
Zinder SH, Cardwell SC, Anguish T, Lee M, Koch M (1984) Methanogenesis in a thermophilic (58°C) anaerobic digestor: Methanothrix sp. as an important aceticlastic methanogen. Appl Environ Microbiol 47:796–807
Zinder SH, Sowers KR, Ferry JG (1985) Methanosarcina thermophila sp. nov., a thermophilic, acetotrophic, methane-producing bacterium. Int J Syst Bacteriol 35:522–523
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Murray, P.A., Zinder, S.H. Polysaccharide reserve material in the acetotrophic methanogen, Methanosarcina thermophila strain TM-1: accumulation and mobilization. Arch. Microbiol. 147, 109–116 (1987). https://doi.org/10.1007/BF00415270
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DOI: https://doi.org/10.1007/BF00415270