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  • Thermostable protein  (2)
  • Cell wall-less mutant  (1)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 108 (1976), S. 189-194 
    ISSN: 1432-072X
    Keywords: Chlamydomonas reinhardii ; Synchronous growth ; Cell wall-less mutant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Synchronization and synchronous growth of a cell wall-less mutant of Chlamydomonas reinhardii have been described. The following growth conditions were used: A modified Sueokas' “high salt minimal medium”, 14∶10 h light-dark cycle, growth temperature 30°C, light intensity 12–18 Klux and dilution of the culture at the end of the dark to a constant cell density of 1.0·106 cells/ml. The time course of increase and distribution of cell volume, cytoplasmic and nuclear division, release of motile cells after the division period and accumulation of DNA, RNA and protein are reported. These mutant cells did not make any sporangium in which the dividing cells were kept as a unit inside a mother cell wall. However, they usually adhered during the period of division, thus making clumps containing 2, 4 and 8 cells. Several of these cell clumps dissolved releasing either single or couples of 2 and 4 cells. After the end of division the cells became flagellated and motile and thereby releasing themselves from the aggregate.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-072X
    Keywords: Key words Isocitrate dehydrogenase ; icd ; Archaea ; Archaeoglobus fulgidus ; Thermostable protein ; Thermostability ; Thermophiles ; Hyperthermophiles ; Phylogeny
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A thermostable homodimeric isocitrate dehydrogenase from the hyperthermophilic sulfate-reducing archaeon Archaeoglobus fulgidus was purified and characterized. The mol. mass of the isocitrate dehydrogenase subunit was 42 kDa as determined by SDS-PAGE. Following separation by SDS-PAGE, A. fulgidus isocitrate dehydrogenase could be renatured and detected in situ by activity staining. The enzyme showed dual coenzyme specificity with a high preference for NADP+. Optimal temperature for activity was 90° C or above, and a half-life of 22 min was found for the enzyme when incubated at 90° C in a 50 mM Tricine-KOH buffer (pH 8.0). Based on the N-terminal amino acid sequence, the gene encoding the isocitrate dehydrogenase was cloned. DNA sequencing identified the icd gene as an open reading frame encoding a protein of 412 amino acids with a molecular mass corresponding to that determined for the purified enzyme. The deduced amino acid sequence closely resembled that of the isocitrate dehydrogenase from the archaeon Caldococcus noboribetus (59% identity) and bacterial isocitrate dehydrogenases, with 57% identity with isocitrate dehydrogenase from Escherichia coli. All the amino acid residues directly contacting substrate and coenzyme (except Ile-320) in E. coli isocitrate dehydrogenase are conserved in the enzyme from A. fulgidus. The primary structure of A. fulgidus isocitrate dehydrogenase confirmes the presence of Bacteria-type isocitrate dehydrogenases among Archaea. Multiple alignment of all the available amino acid sequences of di- and multimeric isocitrate dehydrogenases from the three domains of life shows that they can be divided into three distinct phylogenetic groups.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-072X
    Keywords: Key wordsArchaea ; Archaeoglobus fulgidus ; Thermophiles ; Malate dehydrogenase ; Lactate ; dehydrogenase ; Thermostable protein ; mdh ; Glycine motif ; Lactate-dehydrogenase-like malate ; dehydrogenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A thermostable l-malate dehydrogenase from the hyperthermophilic sulfate-reducing archaeon Archaeoglobus fulgidus was isolated and characterized, and its gene was cloned and sequenced. The enzyme is a homodimer with a molecular mass of 70 kDa and catalyzes preferentially the reduction of oxaloacetic acid with NADH. A. fulgidus l-malate dehydrogenase was stable for 5 h at 90° C, and the half-life at 101° C was 80 min. Thus, A. fulgidus l-malate dehydrogenase is the most thermostable l-malate dehydrogenase characterized to date. Addition of K2HPO4 (1 M) increased the thermal stability by 40%. The primary structure shows a high similarity to l-lactate dehydrogenase from Thermotoga maritima and gram-positive bacteria, and to l-malate dehydrogenase from the archaeon Haloarcula marismortui and other l-lactate-dehydrogenase-like l-malate dehydrogenases.
    Type of Medium: Electronic Resource
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