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US 5,330,901 AGranted
Expression of human serum albumin in Pichia pastoris
AssigneeResearch Corporation Technologies, Inc.
InventorsWilliam D. Prevatt, Kotikanyadan Sreekrishna
Life of this patent
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Abstract
A process for the production of HSA in Pichia pastoris cells comprising cultivating Pichia pastoris cells capable of expressing HSA at a pH of about 5.7 to about 6.4 contemporaneously with the expression of HSA.
Claims16
An improved process for the secretion of a HSA protein in transformed Pichia pastoris cells comprising:
- (a) cultivating in a fermentation broth said transformed Pichia pastoris cells which express a HSA structural gene encoding a native HSA secretion signal sequence and a mature HSA protein, wherein the native HSA signal sequence is operably linked to the sequence encoding the mature HSA protein, under conditions suitable for sustaining the viability of said transformed Pichia pastoris cells, under suitable conditions for the expression of said HSA protein by said Pichia pastoris cells, and
- (b) maintaining the pH of said fermentation broth at a pH in the range of from about 5.7 to about 6.4 contemporaneously with the expression of a HSA protein.
An improved process for the secretion of HSA in transformed cells of Pichia pastoris GS115 comprising:
- (a) cultivating in a fermentation broth cells of Pichia pastoris GS115 which have been transformed with a linear integrative site-specific vector containing the following serial arrangement:
- a first insertable DNA fragment 5' AOX1 Promoter at least one expression cassette containing a HSA structural gene encoding a signal sequence and a mature HSA protein operably linked to a Pichia pastoris AOX1 regulatory region and an AOX1 termination sequence, at least one marker gene and a second insertable DNA fragment 3' to AOX1 termination sequence, under conditions suitable for sustaining the viability of said transformed cells of Pichia pastoris GS115, under suitable conditions for the expression of said HSA by said cells of Pichia pastoris GS115, and
- (b) maintaining the pH of said fermentation broth at a pH in the range of from about 5.7 to about 6.4 contemporaneously with the expression of HSA protein.
Dependencies are inferred from the claim text.