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US 2006/0234351 A1Published application
Yeast protein expression secretion system
OwnerMaharaj Sahib, Edupuganti Rajuinventors
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OwnerMaharaj Sahib, Edupuganti Rajuinventors
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This invention discloses novel prepro-insulin polypeptides. The polypeptides consist of an N-terminal region, derived from N-terminal regions of secretory proteins, and a downstream insulin polypeptide region. The N-terminal region directs the polypeptides efficiently into the secretory pathway of yeasts. Modifications at the N-terminal region, just adjacent to the insulin polypeptide region, further increase the efficiency of secretion and improves the final yield of secreted insulin. The patent also discloses expression systems for the expression of said polypeptides under the regulation of yeast derived alcohol inducible promoters. Thus a combination of such promoters and precursors with the said N-terminal regions appear to function as very high yielding expression systems in yeasts.
1) A DNA construct having a formula
12) A polypeptide SP-B(1-29)-A(1-21) B(1-29)-A(1-21), where SP is a signal peptide region that enables the secretion of polypeptides expressed in yeasts and is derived from either Schwanniomyces occidentalis glucoamylase signal peptide sequence or from Carcinus maenas crustacean hyperglycemic harmone signal peptide sequence, and lies to the N-terminus of the insulin peptide region B(1-29)-A(1-21), and further where B(1-29) is the B chain of insulin from amino acid 1 to amino acid 29, A(1-21) is the A chain of insulin from amino acid 1 to amino acid 21, and the amino acid 29 of the B chain directly connects, by means of a peptide bond, the amino acid 1 of the A chain.
Dependencies are inferred from the claim text.