Method of producing non-bovine chymosin and use hereof
AssigneeChr. Hansen A/S
InventorsStefan Kappeler, Zakaria Farah, Johannes Maarten van den Brink, Henrik Rahbek-Nielsen, Peter Budtz
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Abstract
A method of recombinantly producing a non-bovine pre-prochymosin, prochymosin or chymosin derived from ruminant species including deer species, buffalo species, antelope species, giraffe species, ovine species and caprine species; Camelidae species such as Camelus dromedarius; porcine species; or Equidae species. The recombinant enzymes are used in milk coagulating compositions in cheese manufacturing based on cow's milk and milk from any animal species which are used in cheese manufacturing including camel's milk.
Claims48
A method of producing a non-bovine pre-prochymosin, prochymosin or chymosin, the method comprising the steps of
- (i) isolating or constructing a nucleic acid sequence coding for the pre-prochymosin, prochymosin or chymosin,
- (ii) constructing an expression vector comprising said coding sequence and, operably linked thereto, appropriate expression signals permitting the pre-prochymosin, prochymosin or chymosin to be expressed in a host cell,
- (iii) transforming said host cell with the expression vector,
- (iv) cultivating the thus transformed host cell under conditions where the coding sequence is expressed, and
- (v) harvesting the pre-prochymosin, prochymosin or chymosin.
A method according to claim I wherein the coding sequence is derived from a mammalian species selected from the group consisting of a ruminant species, a Camelidae species, a porcine species, an Equidae species and a primate species.
A DNA construct capable of expressing non-bovine pre-prochymosin, prochymosin or chymosin, said construct comprising an expression vector comprising a nucleic acid sequence comprising a gene coding for the pre-prochymosin, prochymosin or chymosin and, operably linked thereto, appropriate expression signals permitting the pre-prochymosin, prochymosin or chymosin to be expressed in a host cell.
A method of manufacturing cheese, comprising adding a milk clotting effective amount of a non-bovine prochymosin or chymosin to the milk and carrying out appropriate further cheese manufacturing steps, the non-bovine prochymosin or chymosin having in said milk a C/P ratio in the range of 2-20.
A milk clotting composition comprising a milk clotting bovine enzyme selected from prochymosin, chymosin and pepsin and a non-bovine milk clotting enzyme selected from prochymosin, chymosin, pepsin and a microbial aspartic protease.
Dependencies are inferred from the claim text.