Export and modification of (poly)peptides in the lantibiotic way
First Claim
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1. A method for harvesting a desired polypeptide produced by a host cell, said method comprising:
- selecting a host cell having first nucleic acid comprising;
a first nucleic acid fragment encoding a leader peptide;
a second nucleic acid fragment encoding the polypeptide;
wherein the first and second nucleic acid fragments are within the same open reading frame as the first nucleic acid; and
wherein the leader peptide is at least functionally equivalent to a N-terminal leader peptide found within a prepeptide of a lantibiotic;
selecting the host cell for the presence of the transporter protein LanT or a functional equivalent thereof, translating the first nucleic acid; and
harvesting the polypeptide from the culture medium of the host cell.
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Abstract
The invention includes a method for harvesting a polypeptide produced by a host cell, wherein the polypeptide has not undergone intra-cellular post-translational modification, such as dehydration of a serine or a threonine, and/or thioether bridge formation. The invention also includes a method for producing thioether containing peptides and dehydroalanine/dehydrobutyrine-containing peptides, wherein extracellularly thioether rings may be formed.
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Citations
13 Claims
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1. A method for harvesting a desired polypeptide produced by a host cell, said method comprising:
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selecting a host cell having first nucleic acid comprising;
a first nucleic acid fragment encoding a leader peptide;
a second nucleic acid fragment encoding the polypeptide;
wherein the first and second nucleic acid fragments are within the same open reading frame as the first nucleic acid; and
wherein the leader peptide is at least functionally equivalent to a N-terminal leader peptide found within a prepeptide of a lantibiotic;
selecting the host cell for the presence of the transporter protein LanT or a functional equivalent thereof, translating the first nucleic acid; and
harvesting the polypeptide from the culture medium of the host cell. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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Specification