Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1993-06-30
1995-02-21
Lilling, Herbert J.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435 712, 435212, 4352521, 435219, C12P 2104, C12N 952, C12N 950
Patent
active
053914899
ABSTRACT:
Cell-free extracts from Pyrococcus furiosus were found to possess unusually high levels of proteolytic activity as measured by hydrolysis of azocasein; loss in activity was only 30% after incubation for 24 hours at 98.degree. C. and the half-life of proteolytic activity at that temperature was about 60 hours. Furthermore, cell-free extracts incubated at 98.degree. C. in 1% sodium dodecyl sulfate (SDS) for 24 hours yielded an SDS-resistant protease having a temperature optimum of at least 100.degree. C. The enzyme retained at least 40% of its activity when tested at 98.degree. C. by azocasein hydrolysis in the presence of 4M urea, 2M guanidinium chloride, 10 mM dithiothreitol or 150 mM .beta.-mercaptoethanol. The protease was found to have a pH optimum of 6.8 at 98.degree. C. and retained more than 45% of its activity at pH 9.3 and 82% of its activity at pH 4.5 in assays performed at those values. The protease was classified as a metalloprotease through inhibitor studies, and peptide hydrolysis showed trypsin-like cleavage with additional activities.
REFERENCES:
patent: 5242817 (1993-09-01), Kelly et al.
Khoo et al Biochem J. (1984) vol. 221 pp. 407-413.
Matsuzawa et al Agric Biol Chem 47(1) pp. 25-28 (1983).
Cowan et al Biochem J. (1987) 247 pp. 121-133.
Bryant et al, "Characterization of hydrogenase from the Hyperthermophilic Archaebacterium, Pyrococcus furiosus", The Journal of Biological Chemistry, vol. 264, No. 264, No. 9, Issue of Mar. 25, pp. 5070-50079.
Aono et al, "A Novel and Remarkably Thermostable Ferrodioxin from the Hyperthermophilic Archaebacterium Pyrococcus furiosus", Journal of Bacteriology, Jun. 1989, pp. 3433-3439, vol. 171, No. 6.
Kelly et al, "Extremely Thermophilic Archaebacteria: Biological and Engineering Considerations", Biotechnology Progress (vol. 4, No. 2, Jun. 1988, reprinted from Chemical Engineering Progress, Aug. 1988, pp. 47-62.
Brown et al, "Cultivation Techniques for Hyperthermophilic Archaebacteria: Continuous Culture of Pyrococcus furiosus at Temperatures near 100 C", Appl'd and Environmental Microbiology, Aug. 1989, vol. 55, No. 8, pp. 2086-2088.
Parameswaran et al, "Engineering Considerations for Growth of Bacteria at Temperatures Around 100C", The Humana Press Inc., Copyright 1988, pp. 53-73.
Cowan et al, "An extremely thermostable extracellular proteinase from a strain of the archaebacterium Desulfurococcus growing at 88 C", Biochem. J. (1987) 247, 121-133, pp. 121-133.
Matsuzawa et al, "Production of Thermophilic Extracellular Proteases (Aqualysins I and II) by Thermus aquaticus YT-1, an Extreme Thermophile", Agric. Biol. Chem. 47 (1), 25-28, 1983.
Khoo et al, "Interactions of calcium and other metal ions with caldolysin, the thermostable proteinase from Thermus aquaticus strain T351", Biochem. J. (1984) 221, 407-413.
DSM Catalogue of Strains 1989, Pyrococcus Furiosus Stetter and Fiale 2986; Media #377 Pyrococcus/Staphylothermus Medium, pp. I-VIII; p. 97.
Zillig et al, Pyrococcus woesei, sp.nov., an Ultra-Thermophilic Marine Archaebacterium, Representing a Novel Order, Thermococcales, System Appl. Micro Biol. 9, 62-70 (1987).
Matsubara, Methods in Enzymology, vol. XIX, Protolytic enzymes, 1970 Academic Press, N.Y. "Purification and Assay of Thermolysin", 642-650.
Fiala et al, "Pyrococcus furiosus sp.nov. represents a novel genus . . . ", Arch microbiol (1986) 145: 56-61.
Crueger et al, "Biotechnology: A textbook of Industrial Microbiology", Brock, ed., pp. 98-99 (1984).
Anfinsen Christian B.
Blumentals Ilse I.
Brown Stephen H.
Kelly Robert M.
Robinson Anne K. S.
Johns Hopkins University
Lilling Herbert J.
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