Thermostable protease from Staphylothermus

Details Thermostable protease from Staphylothermus Thermostable protease from Staphylothermus

1992-10-22

1994-09-13

Naff, David M.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

435212, 435219, 25217412, C12N 950, C12N 952, C12N 948, C11D 1000

Patent

active

053468210

DESCRIPTION:

BRIEF SUMMARY
TECHNICAL FIELD

This invention is within the field of thermostable proteases. More specifically, the present invention relates to novel thermostable proteases, to a process for the preparation of these enzymes, and to detergent compositions comprising these enzymes.


BACKGROUND ART

Hyperthermophilic archaebacteria have been isolated from solfataric and submarine hydrothermal systems (Kelly, R. M. & Deming, J. W.; Biotech. Progress, 4, 47-62 (1988)). It has been presumed that members of Pyrococcus and Thermococcus contain heat stable proteases and amylases (Stetter, K. O.; J. Chem. Technol. Biotechnol., 43(4), 315-317 (1988)), but proteases obtainable from members of Staphylothermus have never been predicted, isolated or in other ways investigated.


BRIEF DISCLOSURE OF THE INVENTION

Within the scope of the present invention novel enzymes that show extraordinary thermostability as well as thermoactivity are provided. Accordingly, in its first aspect, the present invention provides a protease that is characterized by having pH optimum in the range of from pH 6.5 to 10, and temperature optimum in the range of from 90.degree. to 100.degree. C. In another aspect, the present invention provides a protease that is characterized by having pH optimum in the range of from pH 6.5 to 10, temperature optimum in the range 90.degree. to 100.degree. C., and immunochemical properties identical or partially identical to those of the protease derived from Staphylothermus marinus, DSM No. 3639.
In a third aspect, the present invention provides a process for the preparation of the thermostable proteases of the invention, which process comprises cultivation of a protease producing strain of staphylothermus in a suitable nutrient medium, containing carbon and nitrogen sources and inorganic salts, followed by recovery of the desired enzyme. In preferred embodiments of this process, a strain of Staphylothermus marinus, preferably Staphylothermus marinus, DSM No. 3639, or a mutant or a variant thereof, is cultivated.


BRIEF DESCRIPTION OF THE DRAWING

The present invention is further illustrated by reference to the accompanying drawing, in which FIGS. 1A and 1B show the relation between the proteolytic activity of the protease obtained from Staphylothermus marinus, and temperature and pH, respectively.


DETAILED DISCLOSURE OF THE INVENTION

Growth experiments with Staphylothermus have now shown that these organisms secrete extremely thermostable and thermoactive protein hydrolyzing enzymes. These enzymes possess proteolytic activity under extreme conditions. The properties of the Staphylothermus proteases are demonstrated by the protease obtained from Staphylothermus marinus. A strain of Staphylothermus marinus is available from DSM, No. 3639.
As appears from the Figure, the protease obtainable from Staphylothermus marinus is active in a broad temperature and pH range, namely at temperatures of from below 65.degree. C. to above 100.degree. C., and at pH values of from below pH 6 to above 10. The temperature optimum is between 90.degree. and 100.degree. C., around 95.degree. C. Approximately 80% of proteolytic activity is still detected at 100.degree. C. Moreover, it appears from the Figure that the relation between protease activity and pH leads to a flat activity curve, which means that the protease is very pH tolerant, possessing a generally high proteolytic activity over a broad pH range. In this way the proteases according to the invention have pH optimum in the range of from pH 6.5 to 10, more specifically between pH 8 and pH 10, yet more specifically between pH 8.5 and pH 9.5, around pH 9. At pH 6 and 10, respectively, approximately 70% of proteolytic activity is detected.
In table 1 some of the properties of the proteases obtainable from Staphylothermus sp. are shown.


TABLE 1 ______________________________________ Staphylothermus marinus ______________________________________ pH optimum 9.0 temperature optimum 95.degree. C. type serine substrate specificity: Z-DL-Arg-pNA - Suc-Ala-Ala-Pro-Phe-pNA

REFERENCES:
patent: 3796635 (1974-03-01), Delente
patent: 4480036 (1984-10-01), Morgan et al.
Cowan et al. (1987) Biochem. J., 247, 121-133.
Kelly et al. (1988) Biotechnol. Prog., 4(2), 47-62.
Fiala et al. (1986) System, Appl. Microbiol., 8, 106-113.
Cowan et al. (1985) Trends Biotechnol., 3(3), 68-72.
Bragger et al. (1989) Appl. Microbiol. Biotechnol, 31(56) 556-561.
Cowan et al. (1982) Biochem, Biophys Acta, 705, 293-305.
Stetter et al. (1986) Experientia, 42(11/12), 1187-1191.
Fusek et al. (1990) J. Biol. Chem., 265(3), 1496-1501.
Daniel (1992) Origin Life & Evol. in Biosphere, 22, 33-42.
Eggen et al. (1990) FEMs Microbiol. Lett., 71(1/2), 17-20.
Klingeberg et al. (1991) Appl. Microbiol. Biotechnol., 34(6), 715-719.
Blumenthals et al. (1990) Appl. Environ. Microbiol., 56(7), 1992-1998.
Borman (4 Nov. 1991) Chem. Eng. News, 31-34.
Zamost et al., Chem. Abs. No. 117332y, vol. 114, No. 13, p. 308 (1990).
Takii et al., Chem. Abs. No. 127292a, vol. 108, No. 15, p. 332 (1987).
Meito Sangyo Co., Ltd., Chem. Abs. No. 2625z, vol. 82, No. 1, p. 244 (1974).
Inoue et al., Chem. Abs. No. 6098r, vol. 112, No. 1, p. 618 (1990).
Tosh Corp., Patent Abs. of JP No. C551, vol. 12, No. 472 (1988).

Affiliated with

Also associated with

Rating

Thermostable protease from Staphylothermus does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Thermostable protease from Staphylothermus, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Thermostable protease from Staphylothermus will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-1119769