Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1993-11-11
1999-01-12
Wax, Robert A.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435222, 435220, 435 691, 4353201, 4352523, 43525231, 536 232, 935 14, 935 29, 935 74, C12N 954, C12N 1511, C12N 1557, C12N 1575
Patent
active
058587578
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to novel stabilized proteases, nucleotide sequences encoding the stabilized proteases, and host organisms containing the nucleotide sequences encoding the novel stabilized proteases.
BACKGROUND ART
Proteases, or (interchangeably) peptidases, are enzymes that cleave the amide linkages in protein substrates. Bacteria of the Bacillus species secrete two extracellular species of protease, a neutral or metalloprotease, and an alkaline protease which is functionally a serine endopeptidase, referred to as subtilisin.
A serine protease is an enzyme which catalyses the hydrolysis of peptide bonds, and in which there is an essential serine residue at the active Edition, McGraw-Hill Book Company, N.Y., 271-272!.
The bacterial serine proteases have molecular weights in the range of 20,000 to 45,000. They hydrolyse simple terminal esters and are similar in activity to eukaryotic chymotrypsin, also a serine protease. A more narrow term, alkaline protease, covering a sub-group, reflects the high pH see Priest; Bacteriological Rev., 41 711-753 (1977)!.
In relation to the present invention a subtilisin is a serine protease produced by Gram-positive bacteria or fungi. According to another definition, a subtilisin is a serine protease, wherein the relative order of the amino acid residues in the catalytic triad is Asp--His--Ser (positions 32, 64, and 221). A wide variety of subtilisins has been identified, and the amino acid sequences of a number of subtilisins have been determined. These include among others six subtilisins from Bacillus strains, namely, subtilisin 168, subtilisin BPN', subtilisin Carlsberg, subtilisin DY, subtilisin amylosacchariticus, and mesentericopeptidase (1983); Nucleic Acids Res.; 11 7911-7925; Stahl and Ferrari (1984); J. Bacteriol.; 159 811-819; Jacobs et al. (1985); Nucl. Acids Res.; 13 8913-8926; Nedkov et al. (1985); Biol. Chem. Hoppe-Seyler; 366 421-430; Svendsen et al. (1985); FEBS LETTERS; 196 228-232! one subtilisin from an al. (1985); FEBS LETTERS; 1983 195-200!, and one fungal subtilisin, Hoppe-Seyler; 366 584-492!.
Proteases such as subtilisins have found much utility in industry, particularly in detergent formulations, as they are useful for removing proteinaceous stains.
Proteases are globular proteins and quite compact due to the considerable amount of folding of the long polypeptide chain. The polypeptide chain essentially consists of the "backbone" and its "side-groups". As the peptide bond is planar, only rotations around the C.sub.a --N axis and the C.sub.a --C' axis are permitted. Rotation around the C.sub.a --N bond of the peptide backbone is denoted by the torsion angle .phi. (phi), rotation (1984); Proteins; W. H. Freeman and Company, New York!. The choice of the values of these angles of rotation is made by assigning the maximum value of +180.degree. (which is identical to -180.degree.) to the maximally extended chain. In the fully extended polypeptide chain, the N, C.sub.a and C' atoms are all "trans" to each other. In the "cis" configuration, the angles .phi. and .psi. are assigned the value of 0.degree.. Rotation from this position around the bonds so that the atoms viewed behind the rotated bond move "counter-clockwise" is assigned negative values by definition, those "clockwise" are assigned positive values. Thus, the values of the torsion angles lie within the range -180.degree. to +180.degree..
Since the C.sub.a -atoms are the swivel point for the chain, the side-groups (R-groups) associated with the C.sub.a -atoms become extremely important with respect to the conformation of the molecule.
The term "conformation" defines the participation of the secondary and tertiary structures of the polypeptide chains in moulding the overall structure of a protein. The correct conformation of a protein is of prime importance to the specific structure of a protein and contributes greatly to the unique catalytic properties (i.e. activity and specificity) of enzymes and their stability.
The amino acids of polypeptides can be divided into four
REFERENCES:
patent: 4980288 (1990-12-01), Bryan et al.
patent: 5185258 (1993-02-01), Caldwell et al.
patent: 5324653 (1994-06-01), Van Eekelen
patent: 5336611 (1994-08-01), Van Eckelen et al.
patent: 5340735 (1994-08-01), Christianson et al.
Piela et al., Biopolymers, vol. 26, pp. 1587-1600 (1987).
Balajai et al., Biochem. Biophys. Research Comm. vol. 160, No. 1, 109-14 (1989).
Wells et al., Nucleic Acids Research, vol. 11, No. 22, pp. 7911-925 (1983).
Jany et al., Biol. Chem. Hoppe-Seyler, vol. 366, pp. 485-492 (1985).
Meloun et al., FEBS Letters 2463, vol. 183, No. 2, pp. 195-200 (1985).
Svendsen et al., FEBS Letters 3361, vol. 196, No. 2, pp. 228-232 (1986).
Nedkov et al., Biol. Chem. Hoppe-Seyler, vol. 366, pp. 421-430 (1985).
Jacobs et al., Nucleic Acid Research, vol. 13, No. 24, pp. 8913-8926.
Vasantha et al., J. Bacteriology, vol. 159, No. 3, pp. 811-819 (1984).
Kurihara et al., J. Biological Chem., vol. 247, No. 17, pp. 5619-5631 (1985).
Priest, Bacteriological Reviews, vol. 41, No. 3, pp. 711-753 (1977).
White et al. Principles of Biochemistry, Fifth Edition, pp. 271-272 (1973).
Takagi, H., et al., 1989, Journal of Biochemistry, 105(6): 953-956.
Wu, H. et al., 1990, Proceedings of the National Academy of Sciences, U.S.A., 87:5888-5892.
Estink, M.R., et al., 1990, Proceedings of the SPIE International Society of Optical Engineers, 10241(2Pt.1): 137-143.
Branner Sven
Casteleijn Eric
Egmond Maarten Robert
Eriksen Nina
Hedeg.ang.rd Lisbeth
Harrington James J.
Moore William W.
Novo Nordisk A S
Wax Robert A.
Zelson Steve T.
LandOfFree
Stabilized enzymes and detergent compositions does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Stabilized enzymes and detergent compositions, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Stabilized enzymes and detergent compositions will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1514490