Thermotoga neapolitana DSM 5068 alkaline phosphatase (PhoA) gene and recombinant enzyme production
AssigneeMichigan State University
InventorsClaire Vieille, J. Gregory Zeikus, Maris Laivenieks
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Abstract
The present invention relates to compositions and methods for providing a recombinant thermostable Thermotoga neapolitana alkaline phosphatase enzyme. More particularly, the invention relates to engineering Escherichia coli with T. neapolitana alkaline phosphatase gene expression vectors for providing an inducible system for thermostable enzyme production, wherein the expressed enzyme is readily soluble with a high degree of activity. These methods provide for commercial quantities of a thermostable alkaline phosphatase enzyme.
Claims35
An expression vector comprising a nucleic acid at least 78% identical to SEQ ID NO:03 encoding an alkaline phosphatase polypeptide operably linked to an inducible promoter.
A composition comprising a heterologous nucleic acid, wherein said nucleic acid is at least 78% identical to SEQ ID NO:03.
A method for providing commercial quantities of alkaline phosphatase, comprising,
- a) providing,
- i) a microorganism comprising an expression vector, wherein said expression vector comprises a nucleic acid at least 78% identical to SEQ ID NO:03, operably linked to an inducible promoter;
- ii) an inducing agent for said inducible promoter;
- ii) culture media for said microorganism;
- and
- b) contacting said microorganism with an inducing agent for expressing a commercial quantity of alkaline phosphatase polypeptide in said culture media.
Dependencies are inferred from the claim text.