Method of improving acid decarboxylase activity in vitro under alkaline pH
AssigneesCathay Biotech Inc., Cibt America Inc.
InventorsHoward Chou, Ling Chen, Wenqiang Lu, Xiucai Liu
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Abstract
This invention provides acid decarboxylase-prion subunit fusion polypeptides, nucleic acid sequences, expression vectors, and host cells expression such fusion polypeptides to produce various amino acids and derivatives of the amino acids such as polyamines.
Claims6
A method of improving Escherichia coli lysine decarboxylase CadA polypeptide activity by at least 5% in vitro under alkaline pH comprising fusing a prion subunit to the carboxyl terminus of the Escherichia coli lysine decarboxylase CadA polypeptide, wherein the prion subunit has at least 95% identity to the amino acid sequence of the prion subunit region of any one of SEQ ID NOS:
- 7, 8, 11, 12, 13, 14, 15, 17, or 19, and the Escherichia coli lysine decarboxylase CadA polypeptide has the amino acid sequence of SEQ ID NO:
- 2 or a biologically active fragment thereof that has lysine decarboxylase CadA polypeptide activity and has at least 95% amino acid sequence identity to SEQ ID NO:
- 2, and wherein the lysine decarboxylase CadA polypeptide activity is assessed by measuring the production of cadaverine and lysine Produced by a host cell that expresses the lysine decarboxylase cadA polypeptide fused to the prion subunit and a control cell that expresses the acid decarboxylase protein, but is not fused to the prion subunit, under identical conditions.
Dependencies are inferred from the claim text.