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AssigneesKali-Chemie Aktiengesellschaft, Gesellschaft fuer Biotechnologische Forschung mbH
InventorsRoman Vetter, Detlef Wilke, Antoine Amory, Andre Clippe, Dietmar Schomburg, Wolfgang Aehle
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Novel, optimized highly alkaline proteases which are suitable for use in detergent formulations are prepared by employing microorganisms transformed by mutated DNA sequences. The mutated sequences are obtained starting from DNA sequences which code for highly alkaline protease usually produced by Bacillus species by altering these DNA sequences in defined positions by directed mutagenesis (point mutation) in such a way that the codon in which the point mutation is located now codes for an amino acid which is more strongly basic than the original amino acid. The result is highly alkaline proteases in which original amino acids have been replaced by more strongly basic amino acids, preferably by the amino acids lysine or arginine. Synthetic oligonucleotides, DNA sequences, vectors and transformed microorganisms which are used for generating and obtaining the optimized highly alkaline protease are also described.
Highly alkaline protease, which has an amino-acid sequence having at least 80% sequence similarity with the protease amino-acid sequence illustrated in FIG. 1 (SEQ. ID. NO. 2) commencing at residue number 1 at the `protease` mark and differing from said FIG. 1 (SEQ. ID. NO. 2) sequence in from one to three of the positions 18, 42, 49, 57, 96, 107, 114, 115, 135, 138, 166, 176, 179, 188, 189, 198, 238, 255, or 266 in FIG. 1 (SEQ. ID. NO. 2), or sequence positions corresponding thereto, in that the amino acid located in the differing position has been replaced by a more strongly basic amino acid selected from the group consisting of lysine and arginine.
Highly alkaline protease, which has an amino-acid sequence having at least 80% sequence similarity with the protease amino-acid sequence illustrated in FIG. 1 (SEQ. ID. NO. 2) commencing at residue number 1 at the `protease` mark and differing from said FIG. 1 (SEQ. ID. NO. 2) sequence in from one to three of the positions 18, 49, 57, 107, 114, 115, 135, 138, 166, 176, 179, 188, 189, 198, 238, or 255 in FIG. 1 (SEQ. ID. NO. 2), or sequence positions corresponding thereto, in that the amino acid located in the differing position has been replaced by a more strongly basic amino acid selected from the group consisting of lysine and arginine.
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