Variants of chymosin with improved milk-clotting properties
AssigneeChr. Hansen A/S
InventorsJohannes Maarten Van Den Brink, Martin Lund, Jonas Jacobsen, Sari Charlotte Hansen, Iben Jeppesen
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Abstract
Variants of chymosin with improved milk-clotting properties.
Claims20
A method for making an isolated chymosin polypeptide variant comprising:
- (a) producing a chymosin polypeptide variant having an alteration at one or more positions in its amino acid sequence relative to a parent polypeptide having chymosin activity, wherein the alteration comprises a substitution, a deletion or an insertion in at least one amino acid position corresponding to any of positions 117, 134, 141, 143, 156, 241, 279, 280, 281, 298, 300, 307, 309, 311, 325, 350, 352 and 353;
- and
- (b) isolating the polypeptide variant of step (a), thereby obtaining the isolated chymosin polypeptide variant, wherein the variant has chymosin activity;
- and wherein:
- (i):
- the amino acid position of the parent polypeptide is determined by an alignment of the amino acid sequence of the parent polypeptide with the amino acid sequence of the polypeptide of SEQ ID NO:
- 1 (bovine chymosin);
- and
- (ii):
- the parent polypeptide has at least 65% sequence identity with the mature polypeptide from amino acid position 59 to amino acid position 381 of SEQ ID NO:
- 1 (bovine chymosin).
An isolated chymosin polypeptide variant having an alteration at one or more positions in its amino acid sequence relative to a parent polypeptide having chymosin activity, wherein the alteration comprises a substitution, a deletion or an insertion in at least one amino acid position corresponding to any of positions 117, 134, 141, 143, 156, 241, 279, 280, 281, 298, 300, 307, 309, 311, 325, 350, 352 and 353, wherein the variant has chymosin activity;
- and wherein:
- (i):
- the amino acid position of the parent polypeptide is determined by an alignment of the amino acid sequence of the parent polypeptide with the amino acid sequence of the polypeptide of SEQ ID NO:
- 1 (bovine chymosin);
- and
- (ii):
- the parent polypeptide has at least 90% sequence identity with the mature polypeptide from amino acid position 59 to amino acid position 381 of SEQ ID NO:
- 1 (bovine chymosin),
- (iii):
- the isolated chymosin polypeptide variant has less than 100% sequence identity with the mature polypeptide of SEQ ID NO:
- 1 (bovine chymosin);
- and
- wherein the isolated chymosin polypeptide variant has a chymosin activity giving a higher C/P ratio as compared to the C/P ratio of bovine chymosin comprising the mature polypeptide of SEQ ID NO:
- 1.
An isolated chymosin polypeptide variant having an alteration at one or more positions in its amino acid sequence relative to a parent polypeptide having chymosin activity, wherein the alteration comprises a substitution, a deletion or an insertion in at least one amino acid position corresponding to any of positions 117, 134, 141, 143, 156, 241, 279, 280, 281, 298, 300, 307, 309, 311, 325, 350, 352 and 353, wherein the variant has chymosin activity;
- and wherein:
- (i):
- the amino acid position of the parent polypeptide is determined by an alignment of the amino acid sequence of the parent polypeptide with the amino acid sequence of the polypeptide of SEQ ID NO:
- 1 (bovine chymosin)
- (ii):
- the parent polypeptide has at least 90% sequence identity with the mature polypeptide from amino acid position 59 to amino acid position 381 of SEQ ID NO:
- 2 (Camel chymosin);
- and
- (iii):
- the isolated chymosin polypeptide variant has less than 100% sequence identity with the mature polypeptide of SEQ ID NO:
- 2 (camel chymosin);
- wherein the isolated chymosin polypeptide variant has a chymosin activity giving a higher C/P ratio as compared to the C/P ratio of camel chymosin comprising the mature polypeptide of SEQ ID NO:
- 2.
Dependencies are inferred from the claim text.