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Anita Sellstedt - Energy Production using Microorganisms Print E-mail

info_16x16The aim of our research is to study energy production in microorganisms. We use an integrative approach with a combination of molecular biology, biochemistry and ecophysiology.
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Hydrogen metabolism of Frankia
The nitrogen-fixing filamentous actinobacteria of the genus Frankia form root nodules in symbioses with many woody trees and shrubs. Nitrogen fixation by actinorhizal plants may contribute as much as a quarter of the total biologically fixed nitrogen globally in terrestrial ecosystems each year. In addition, actinorhiza are major sources of new soil nitrogen in many different biomes.
The most important recent achievement in the Frankia research field is undoubtedly the sequencing of three Frankia genomes. We were able to show that there are large differences in the genome sizes. Frankia EANpec1 was found to have the largest genome with 9.0 Megabases (Mb), while Frankia ACN14a had an intermediate size (7.5 Mb) and Frankia HFPCcI3 was the smallest at 5.4 Mb. These numbers were correlated with geographical origin, host plant distribution and repeated sequences, such as IS. Our findings open up a new era in Frankia research, yielding possibilities to explore the molecular biology of Frankia and to utilize its traits, e.g. nitrogen fixation.
An inevitable source of energy-inefficiency in the nitrogen-fixation process is the evolution of hydrogen; as much as 25% of the in vitro electron flow through nitrogenase goes to hydrogen evolution, while in vivo these losses are much higher. Some nitrogen-fixing systems have dealt with this problem of energy loss through an enzyme, called uptake hydrogenase, which is very common in Frankia.
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Light micrograph of the bacterium Frankia showing vesicles. Light micrograph of our isolate of Chalara parvispora

Ethanol production
We are interested in developing ethanol production from cellulose-based biomass. We have shown in a batch fermentation study that a fungal mix could produce 24 gL-1 ethanol using pulp waste as substrate. The fungal mix was able to grow on xylose, hemicellulose and cellulose. In addition, we were able to identify the fungi mix using PCR-amplification of DNA and sequencing as Chalara parvispora and Trametes hirsuta/T. versicolor. In a reconstitution study, the identified fungi were shown to produce equal amounts of ethanol as the fungi mix. In addition, it was shown that C. parvispora could produce ethanol from xylose.
The study shows that refining biomass by ethanol production from pulp waste, a lignocellulose material, can be increased by adding C. parvispora and T. versicolor, which is of great economic significance for energy production.
sweden_greySvensk sammanfattning

Key publications

DeLuca TH, O Zackrisson, M-C Nilsson, A Sellstedt 2002. Quantifying nitrogen fixation in feather moss carpets. Nature. 419: 917-920.

Mohapatra A, Leul M, Mattsson U, Sellstedt A 2004. A hydrogen-evolving enzyme is present in Frankia R43. FEMS Micro- biol. Lett.236: 235-240

Leul M, Mattsson U, Sellstedt A 2005. Molecular characterization of uptake hydrogenase in Frankia. Biochem. Soc. Trans. 33: 64-66.

Normand, Lapierre, Tisa, Lavire, Alloisio, Cournoyer, Marechal, Pujic, Gogarten,Huang, Mastronunzio, Bickhard, Bassi, Rawnsley, Niemann, Francino, Lapidus, Martinez, Goltsman, Choisne, Schenowitz, Couloux, Demange, Medigue, Cruveiller, Labarre, Rouy, Vallenet, Mullin, Kopp, Wang, Tomkins, Berry, Valverde, Wall, Sellstedt, Tavares Daubi, Benson 2007. Genome structure reflects host biogeography in three plant symbionts Frankia sp. strains. Genome Research, 17, 7-15.

Leul M, Normand P, Sellstedt A 2009. The phylogeni of uptake hydrogenases. Int Microbiol. 12(1):23-28.

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  1. Leul M, Normand P, Sellstedt A
    The phylogeny of uptake hydrogenases in Frankia
    International Microbiology: 2009 12:23-28
  2. Zackrisson O, DeLuca TH, Gentili F, Sellstedt A, Jäderlund A
    Nitrogen fixation in mixed Hylocomium splendens moss communities
    Oecologia: 2009 160:309-319
  3. Holmgren M, Sellstedt A
    Identification of white-rot and soft-rot fungi increasing ethanol production from spent sulfite liquor in co-culture with Saccharomyces cerevisiae
    Journal of Applied Microbiology: 2008 105:134-140
  4. Holmgren M, Sellstedt A
    Identification of white-rot and soft-rot fungi increasing ethanol production from spent sulfite liquor in co-culture with Saccharomyces cerevisiae
    Journal of Applied Microbiology: 2008 105:134-140
  5. Leul M, Normand P, Sellstedt A
    The organization, regulation and phylogeny of uptake hydrogenase genes in Frankia

    Physiologia Plantarum: 2007 130:464-470
  6. Santos CL, Vieira J, Sellstedt A, Normand P, Moradas-Ferreira P, Tavares F
    Modulation of Frankia alni ACN14a oxidative stress response: activity, expression and phylogeny of catalases
    Physiologia Plantarum: 2007 130:454-463
  7. Tavares F, Santos CL, Sellstedt A
    Reactive oxygen species in legume and actinorhizal nitrogen-fixing symbioses: the microsymbiont's responses to an unfriendly reception
    Physiologia Plantarum: 2007 130:344-356
  8. Sellstedt A, Normand P, Dawson J
    Frankia - the friendly bacteria - infecting actinorhizal plants
    Physiologia Plantarum: 2007 130:315-317
  9. DeLuca TH, Zackrisson O, Gentili F, Sellstedt A, Nilsson M-C
    Ecosystem controls on nitrogen fixation in boreal feather moss communities
    Oecologia: 2007 152:121-130
  10. Normand P, Lapierre P, Tisa LS, Gogarten JP, ..., Sellstedt A, Tavares F, et al
    Genome characeristics of facultatively symbiotic Frankia sp. strains reflect host range and host plant biogeography
    Genome Research 2007 17:7-15
  11. Leul M, Mohapatra A, Sellstedt A
    Biodiversity of hydrogenases in Frankia
    Curr Microbiol: 2005 50:17-23
  12. Stahl L, Hogberg P, Sellstedt A, Buresh RJ
    Measuring nitrogen fixation by Sesbania sesban planted fallows using N-15 tracer technique in Kenya
    Agroforestry Systems: 2005 65:67-79
  13. Leul M, Mattsson U, Sellstedt A
    Molecular characterization of uptake hydrogenase in Frankia
    Biochem Soc Trans: 2005 33:64-66
  14. Gentili F, Nilsson MC, Zackrisson O, DeLuca TH, Sellstedt A
    Physiological and molecular diversity of feather moss associative N-2-fixing cyanobacteria
    Journal Of Experimental Botany: 2005 56:3121-3127
  15. Mohapatra A, Leul M, Mattsson U, Sellstedt A
    A hydrogen-evolving enzyme is present in Frankia sp. R43
    FEMS Microbiol Lett: 2004 236:235-240
  16. Zackrisson O, DeLuca TH, Nilsson MC, Sellstedt A, Berglund LM
    Nitrogen fixation increases with successional age in boreal forests
    Ecology: 2004 85:3327-3334
  17. Tavares F, Bernardo L, Sellstedt A
    Identification and expression studies of a catalase and a bifunctional catalase-peroxidase in Frankia strain R43
    Plant and Soil: 2003 254:75-81
  18. Leitz G, Lundberg C, Fallman E, Axner O, Sellstedt A
    Laser-based micromanipulation for separation and identification of individual Frankia vesicles
    Fems Microbiology Letters: 2003 224:97-100
  19. Mattsson U, Sellstedt A
    Nickel affects activity more than expression of hydrogenase protein in Frankia
    Curr Microbiol: 2002 44:88-93
  20. DeLuca TH, Zackrisson O, Nilsson M-C, Sellstedt A
    Quantifying nitrogen-fixation in feather moss carpets of boreal forests
    Nature: 2002 419:917-920
  21. Tavares F, Sellstedt A
    DNase-resistant DNA in the extracellular and cell wall-associated fractions of Frankia strains R43 and CcI3
    Curr Microbiol: 2001 42:168-172
  22. Mattsson U, Johansson L, Sandstrom G, Sellstedt A
    Frankia KB5 possesses a hydrogenase immunologically related to membrane-bound
    Curr Microbiol: 2001 42:438-441
  23. Mattsson U, Johansson L, Sandstrom G, Sellstedt A
    Frankia KB5 possesses a hydrogenase immunologically related to membrane-bound [NiFe]-hydrogenases
    Current Microbiology: 2001 42:438-441
  24. Tavares F, Sellstedt A
    A simple, rapid and non-destructive procedure to extract cell wall-associated proteins from Frankia
    J Microbiol Methods: 2000 39:171-178
  25. Mattsson U, Sellstedt A
    Hydrogenase in Frankia KB5: expression of and relation to nitrogenase
    Can J Microbiol: 2000 46:1091-1095
  26. Tavares F, Sellstedt A
    DNase activities of the extracellular, cell wall-associated, and cytoplasmic protein fractions of Frankia strain R43
    Applied and Environmental Microbiology: 1997 63:4597-4599
  27. Sellstedt A
    Specificity and Effectivity in Nodulation by Frankia on Southern-Hemisphere Actinorhiza
    Fems Microbiology Letters: 1995 125:231-236
  28. Sellstedt A, Rosbrook PA, Kang L, Reddell P
    Effect of Carbon Source on Growth, Nitrogenase and Uptake Hydrogenase Activities of Frankia Isolates from Casuarina Sp
    Plant and Soil: 1994 158:63-68
  29. Sellstedt A, Mattsson U
    Hydrogen Metabolism in Casuarina-Frankia - Immunolocalization of Nitrogenase and Hydrogenase
    Soil Biology & Biochemistry: 1994 26:583-592
  30. Sellstedt A, Stahl L, Mattsson U, Jonsson K, Hogberg P
    Can the N-15 Dilution Technique Be Used to Study N-2 Fixation in Tropical Tree Symbioses as Affected by Water-Deficit
    Journal of Experimental Botany: 1993 44:1749-1755
  31. Sellstedt A
    Hydrogen Metabolism in Frankia - Is There a Plant Factor Affecting This Metabolism
    Acta Oecologica-International Journal of Ecology: 1992 13:512-512
  32. Sellstedt A, Wullings B, Nystrom U, Gustafsson P
    Identification of Casuarina-Frankia Strains by Use of Polymerase Chain-Reaction (Pcr) with Arbitrary Primers
    Fems Microbiology Letters: 1992 93:1-5
  33. Sellstedt A, Atkins CA
    Composition of Amino-Compounds Transported in Xylem of Casuarina Sp
    Journal of Experimental Botany: 1991 42:1493-1497
  34. Sellstedt A, Reddell P, Rosbrook P
    The Occurrence of Hemoglobin and Hydrogenase in Nodules of 12 Casuarina-Frankia Symbiotic Associations
    Physiologia Plantarum: 1991 82:458-464
  35. Sellstedt A, Reddell P, Rosbrook PA, Ziehr A
    The Relations of Hemoglobin and Lignin-Like Compounds to Acetylene-Reduction in Symbiotic Casuarina
    Journal of Experimental Botany: 1991 42:1331-1337
  36. Sellstedt A, Winship LJ
    Acetylene, Not Ethylene, Inactivates the Uptake Hydrogenase of Actinorhizal Nodules During Acetylene-Reduction Assays
    Plant Physiology: 1990 94:91-94
  37. Sellstedt A, Lindblad P
    Activities, Occurrence, and Localization of Hydrogenase in Free-Living and Symbiotic Frankia
    Plant Physiology: 1990 92:809-815
  38. Sellstedt A, Smith GD
    Nickel Is Essential for Active Hydrogenase in Free-Living Frankia Isolated from Casuarina
    Fems Microbiology Letters: 1990 70:137-140
  39. Lindblad P, Sellstedt A
    Occurrence and Localization of an Uptake Hydrogenase in the Filamentous Heterocystous Cyanobacterium Nostoc Pcc-73102
    Protoplasma: 1990 159:9-15
  40. Sellstedt A, Hogberg P, Jonsson K
    Diurnal-Variation in Acetylene-Reduction and Net Hydrogen Evolution in 5 Tropical and Sub-Tropical Nitrogen-Fixing Tree Symbioses
    Journal of Experimental Botany: 1989 40:1163-1168
  41. Lindblad P, Sellstedt A
    Immunogold Localization of Hydrogenase in Free-Living Frankia Cpi1
    Fems Microbiology Letters: 1989 60:311-315
  42. Sellstedt A
    Occurrence and Activity of Hydrogenase in Symbiotic Frankia from Field-Collected Alnus-Incana
    Physiologia Plantarum: 1989 75:304-308
  43. Sellstedt A
    Nitrogenase Activity, Hydrogen Evolution and Biomass Production in Different Casuarina Symbioses
    Plant and Soil: 1988 105:33-40
  44. Sellstedt A, Winship LJ
    Hydrogen Metabolism of Casuarina Root-Nodules - a Comparison of 2 Inoculum Sources
    Physiologia Plantarum: 1987 70:367-372
  45. Winship LJ, Martin KJ, Sellstedt A
    The Acetylene-Reduction Assay Inactivates Root Nodule Uptake Hydrogenase in Some Actinorhizal Plants
    Physiologia Plantarum: 1987 70:361-366
  46. Sellstedt A
    Acetylene reduction, H-2 Evolution and 15-N-2 Fixation in the Alnus incana-Frankia Symbiosis
    Planta: 1986 167:382-386
  47. Sellstedt A, Hussdanell K
    Biomass Production and Nitrogen-Utilization by Alnus-Incana When Grown on N-2 or Nh4+ Made Available at the Same Rate
    Planta: 1986 167:387-394
  48. Sellstedt A
    Nitrogen and Carbon Utilization in Alnus-Incana Fixing N-2 or Supplied with No3 at the Same Rate
    Journal of Experimental Botany: 1986 37:786-797
  49. Sellstedt A, Hussdanell K, Ahlqvist AS
    Nitrogen-Fixation and Biomass Production in Symbioses between Alnus-Incana and Frankia Strains with Different Hydrogen Metabolism
    Physiologia Plantarum: 1986 66:99-107
  50. Huss-Danell K, Sellstedt A
    Nitrogenase activity in response to darkening and defoliation in Alnus incana

    Journal of Experimental Botany: 1985 36:1352-1358
  51. Sellstedt A, Huss-Danell K
    Growth, nitrogen fixation and relative efficiency of nitrogenase in Alnus incana grown in different cultivation systems
    Plant and Soil: 1984 78:147-158

 
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