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  • 1
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Extracellular pullulanase (pullulan 6-glucanohydrolase, EC 3.2.1.41) was purified from cell free culture supernatants of Thermoanaerobium Tok6-B1 by ammonium sulphate precipitation, affinity precipitation, gel exclusion and ion exchange chromatography. A final purification factor of over 1600 was achieved. A molecular weight of 120 kD was determined by steric exclusion HPLC. Enzyme activity was specifically directed towards the α 1–6 glucosidic linkages of pullulan resulting in 100% conversion to maltotriose and also possessed activity towards α 1–4 linkages of starch, amylopectin and amylose producing maltooligosaccharides (DP2-DP4) as products. Maltotetraose was slowly hydrolysed to maltose. Values of K m (% w/v) were 7.3×10-3 for pullulan, 2.7×10-3 for amylopectin and 4.7×10-3 for Lintner's starch. Pullulanase activity was resistant to 6 M urea and was thermostable at temperatures up to 80°C (t 1/2 in the order of hours). Above 80°C thermal denaturation was significant (t 1/2=17 min at 85°C; 5 min at 90°C) but became less so in the presence of substrate (pullulan or starch). Thermostability was greatest at the pH activity optimum (pH 5.5) and was promoted by Ca2+ ions.
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  • 2
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary An endo-gb-1,4-mannanase cloned from “caldocellum saccharolyticum” and expressed in Escherichia coli was partially purified. The purification involved heat treatment, anion exchange and gel filtration. The mannanase was only active against mannan, glucomannans and galactoglucomannans and obeyed Michaelis-Menten kinetics on these substrates. The rate and extent of hydrolysis was dependent on the type of substrate. Galactomannans were not as readily depolymerized as the mannan and glucomannans investigated. The glucose content of the glucomannans did not affect the rate of hydrolysis and only slightly affected the extent. The molecular mass of the mannanase was estimated at 39 kDa. The pH and temperature optima were 6.5 and 80° C respectively. The mannanase was very thermostable with a half life of 48 min at 85° C and no loss in activity after 24 h at 70° C.
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  • 3
    ISSN: 1432-1432
    Keywords: Botryoid ; Chemiosmosis ; Ferredoxin ; Greigite ; Hydrothermal ; Iron sulphides ; Membrane ; Origin of life ; Protocell ; Proton motive force
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We propose that life emerged from growing aggregates of iron sulphide bubbles containing alkaline and highly reduced hydrothermal solution. These bubbles were inflated hydrostatically at sulphidic submarine hot springs sited some distance from oceanic spreading centers four billion years ago. The membrane enclosing the bubbles was precipitated in response to contact between the spring waters and the mildly oxidized, acidic and iron-bearing Hadean ocean water. As the gelatinous sulphide bubbles aged and were inflated beyond their strength they budded, producing contiguous daughter bubbles by the precipitation of new membrane. [Fe2S2]+/0 or [Fe4S4]2+/+ clusters, possibly bonded by hydrothermal thiolate ligands as proferredoxins, could have catalyzed oxidation of thiolates to disulphides, thereby modifying membrane properties. We envisage the earliest iron sulphide bubbles (pro botryoids) first growing by hydrostatic inflation with hydrothermal fluid, but evolving to grow mainly by osmosis (the protocellular stage), driven by (1) catabolism of hydrothermal abiogenic organics trapped on the inner walls of the membrane, catalyzed by the iron sulphide clusters; and (2) cleavage of hydrophobic compounds dissolved in the membrane to hydrophilic moieties which were translocated, by the proton motive force inherent in the acidic Hadean ocean, to the alkaline interior of the protocell. The organics were generated first within the hydrothermal convective system feeding the hot springs operating in the oceanic crust and later in the pyritizing mound developing on the sea floor, as a consequence of the reduction of CO, CO2, and formaldehyde by Fe2+- and S2−-bearing minerals. We imagine the physicochemical interactions in and on the membrane to have been sufficiently complex to have engendered auto- and cross-catalytic replication. The membrane may have been constructed in such a way that a “successful” parent could have “informed” the daughters of membrane characteristics functional for the then-current level of evolution.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular evolution 40 (1995), S. 559-563 
    ISSN: 1432-1432
    Keywords: Archaebacteria ; Evolution of metabolism ; Nicotinamide coenzymes ; Nonhem iron proteins ; Redox coenzymes ; Thermophiles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP) are of universal occurrence in living organisms and play a central role in coupling oxidative with reductive reactions. However, the evidence that the origin and early evolution of life occurred at high temperatures (〉95°C) is now strong, and at these temperatures some modern metabolites, including both the reduced and oxidized forms of these coenzymes, are unstable. We believe there is good evidence that indicates that in the most primitive organisms nonhem iron proteins carried out many or all of the functions of NAD/P(H). This has important implications for the way in which investigations of archaebacterial metabolism are conducted.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Extremophiles 3 (1999), S. 263-267 
    ISSN: 1433-4909
    Keywords: Key words Xylanase ; Xylanolytic ; Archaea ; Thermococcus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A xylanase has been found in the archaeon Thermococcus zilligii strain AN1 (DSM 2770), which grows optimally at 75°C. The enzyme had a molecular mass of 95 kDa and a unique N-terminal sequence. It had activity against all five xylans tested and against xylose oligomers, but not against other carbohydrate polymers. The K m values found for xylans were typical of those found for bacterial xylanases. The pH optimum for activity was pH 6, and the enzyme half-life at 100°C was 8 min. This is the first description of any archaeal hemicellulase.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Applied microbiology and biotechnology 34 (1991), S. 789-793 
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Thermus sp. Rt41A produces an extracellular proteinase that is produced concomitant with growth and l-glutamate catabolism. Calcium-chelating medium components were shown to decrease the half-life of the proteinase in growing cultures. Medium modifications avoiding these components resulted in an increase in the half-life and in the peak level of proteinase. By adding the inorganic phosphate requirement for growth in anabolic amounts to pH-controlled batch cultures, stability of the proteinase in the medium was greatly enhanced and there was consequent improvement in the total proteinase yield. This approach also allowed a balanced increase in substrate and phosphate concentrations to increase the cell and proteinase yield in batch culture in an almost stoichiometric manner.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Origins of life and evolution of the biospheres 22 (1992), S. 33-42 
    ISSN: 1573-0875
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Origins of life and evolution of the biospheres 22 (1992), S. 181-190 
    ISSN: 1573-0875
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Type of Medium: Electronic Resource
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