Unglycosylated lysostaphin variant protein
Unglycosylated lysostaphin variant protein, nucleic acid molecule, vector and host cell, as well as a method for production of unglycosylated lysostaphin variant protein in a yeast expression system are provided. The proteins are produced in a Pichia pastoris expression system and have been shown to have activity equivalent to wild-type lysostaphin. The lysostaphin variant proteins can be used as therapeutic proteins for treatment of diseases such as Staphylococcus aureus infection.
1. A vector comprising a recombinant nucleic acid molecule encoding a lysostaphin protein or variant thereof, wherein the recombinant nucleic acid molecule encoding the lysostaphin protein or variant thereof comprises the nucleotide sequence of SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7, and wherein the lysostaphin variant protein has endopeptidase activity.
2. The vector of claim 1 , wherein said vector is an expression vector.
3. The vector of claim 1 , wherein the recombinant nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO:5 and encodes a lysostaphin variant protein comprising the amino acid sequence of SEQ ID NO:4.
4. The vector of claim 3 , wherein said vector is an expression vector.
5. The vector of claim 1 , wherein the recombinant nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO:7 and encodes a lysostaphin variant protein comprising the amino acid sequence of SEQ ID NO:6.
6. The vector of claim 5 , wherein said vector is an expression vector.
7. A recombinant host cell comprising the vector of claim 1 .
8. The recombinant host cell of claim 7 , wherein said host cell is a yeast organism.
9. The recombinant host cell of claim 8 , wherein the yeast organism is Pichia pastoris.
10. The recombinant host cell of claim 8 , wherein said vector is an expression vector, and wherein the recombinant nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO:5 and encodes a lysostaphin variant protein comprising the amino acid sequence of SEQ ID NO:4.
11. The recombinant host cell of claim 10 , wherein the yeast organism is Pichia pastoris.
12. The recombinant host cell of claim 10 , wherein the encoded lysostaphin variant protein is unglycosylated at the position corresponding to N125 of SEQ ID NO:4, and wherein the lysostaphin variant protein has endopeptidase activity.
13. The recombinant host cell of claim 11 , wherein the encoded lysostaphin variant protein is unglycosylated at the position corresponding to N125 of SEQ ID NO:4, and wherein the lysostaphin variant protein has endopeptidase activity.
14. The recombinant host cell of claim 8 , wherein said vector is an expression vector, and wherein the recombinant nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO:7 and encodes a lysostaphin variant protein comprising the amino acid sequence of SEQ ID NO:6.
15. The recombinant host cell of claim 14 , wherein the yeast organism is Pichia pastoris.
16. The recombinant host cell of claim 14 , wherein the encoded lysostaphin variant protein is unglycosylated at the position corresponding to N125 of SEQ ID NO:6, and wherein the lysostaphin variant protein has endopeptidase activity.
17. The recombinant host cell of claim 15 , wherein the encoded lysostaphin variant protein is unglycosylated at the position corresponding to N125 of SEQ ID NO:6, and wherein the lysostaphin variant protein has endopeptidase activity.