Intrazelluläre Proteasen aus <i>Thermoactinomyces vulgaris</i> 1. Mitt. Vorkommen der Proteasen in der Zelle und Dynamik ihrer Bildung im Verlauf der Kulturführung

The intracellular proteases (IP) of the cellular extract of Thermoactinomyces vulgaris are characterized as to the biochemical properties compared with the corresponding extracellular proteases (EP). According to that, the storage stability and the temperature and pH behaviour (optimum, stability) of both of the proteases are identical. Nevertheless, differences were detected between IP and EP after the action of several effectors and different substrates. As could be seen after a column chromatographic separation of the IP of the cellular extract, it is composed of at least 3 proteases, two of them are serine proteases which can be inhibited moreover unspecifically by p-chloromercuri benzoate. The purified proteases (IP) are very instable and therefore not yet characterized in detail.

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“…Mitt. [5]), beim EP-Praparat durch Lyophilisieren der aus dem Kulturfiltrdt mit 60 bis 80 "/, Ethanol ausfillbaren Fraktion.…”

Section: Enzympruparute (Rohenzyme)unclassified

“…1. Mitt[5].Sfrepromycinsulfur-FSiNung. Der Zellextrakt wird zur Entfernung der Nucleinsauren mit Streptomycinsulfat (5 mg/ml) versetzt und der gebildete Niederschlag abgetrennt und verworfen.…”

unclassified

Intrazelluläre Proteasen aus Thermoactinomyces vulgaris 2. Mitt. Biochemische Charakterisierung der Proteasen des Zellextraktes

Schalinatus

Ruttloff

Behnké

1983

Nahrung

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Identification and characterization of a novel spore-associated subtilase from Thermoactinomyces sp. CDF

et al. 2009

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A gene encoding a spore-associated subtilase, designated protease CDF, was cloned from Thermoactinomyces sp. CDF and expressed in Escherichia coli. The enzyme gene is translated as a proform consisting of a 94 aa propeptide and a 283 aa mature protease domain. Phylogenetic analysis revealed that this enzyme belonged to the subtilisin family, but could not be grouped into any of its six known subfamilies. The mature protease CDF has an unusually high content of charged residues, which are mainly distributed on the enzyme surface. The recombinant proform of protease CDF formed inclusion bodies, but could be efficiently converted to the mature enzyme when the inclusion bodies were dissolved in alkaline buffers. The proform underwent a two-step maturation process, wherein the N-terminal part (85 residues) of the propeptide was autoprocessed intramolecularly, and the remaining 9-residue peptide was further processed intermolecularly. Protease CDF exhibited optimal proteolytic activity at 50-55 6C and pH 10.5-11.0. The enzyme was stable under high-pH conditions (pH 11.0-12.0), and NaCl could stabilize the enzyme at lower pH values. In addition, the enzyme was not dependent on calcium for either maturation or stability. By immunoblot analysis, protease CDF was found to be associated with spores, and could be extracted from the spores with 2 M KCl and alkaline buffers without damaging the coat layer, demonstrating that the protease CDF is located on the surface of the spore coat.

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Intrazelluläre Proteasen aus Thermoactinomyces vulgaris 1. Mitt. Vorkommen der Proteasen in der Zelle und Dynamik ihrer Bildung im Verlauf der Kulturführung

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References 10 publications

Intrazelluläre Proteasen aus Thermoactinomyces vulgaris 2. Mitt. Biochemische Charakterisierung der Proteasen des Zellextraktes

Intrazelluläre Proteasen aus Thermoactinomyces vulgaris 2. Mitt. Biochemische Charakterisierung der Proteasen des Zellextraktes

Identification and characterization of a novel spore-associated subtilase from Thermoactinomyces sp. CDF

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