Modified bacterial alkaline phosphatases and their applications

Details Modified bacterial alkaline phosphatases and their applications Modified bacterial alkaline phosphatases and their applications

1996-06-28

1999-04-06

Wax, Robert A.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

4351721, 4351723, 435 691, 4352523, 4353201, 435849, 536 231, 536 237, C12N 914, C12N 1500, C12N 120, C12P 2106

Patent

active

058916997

ABSTRACT:
Modified alkaline phosphatases of bacterial origin (BAP or bacterial alkaline phosphatase) which consist of a bacterial alkaline phosphatase sequence in which at least one of the amino acid residues in position 329 or in position 330 is replaced by another amino acid residue, which modified bacterial alkaline phosphatases exhibit both significantly improved enzymic properties and an increased thermal stability, and their applications, in particular in immunoenzymic assays (reagents and diagnostic kits).
Method for selecting mutants of alkaline phosphatases which possess significantly improved enzymic properties.

REFERENCES:
patent: 5362644 (1994-11-01), Boquet et al.
patent: 5534223 (1996-07-01), Boquet et al.
Claude M.L. Janeway et al., "Magnesium in the Active Site of Escherichia coli Alkaline Phosphatase is Important for Both Structural Stabilization and Catalysis", Biochemistry, vol. 32, pp. 160-1609, 1993.
Daniel Gillet et al., Insertion of a Disulfide-Containing Neurotoxin into E. coli Alkaline Phosphatase: The Hybrid Retains Both Biological Activities, Protein Engineering, vol. 5, No. 3, pp. 273-278, 1992.
J.E. Murphy et al., "Why are Mammalian Alkaline Phosphatase Much More Active than Bacterial Alkaline Phosphatases", Molecular Microbiology, vol. 12, No. 3, 1994.
Janeway et al. (1993) Biochemistry, 32:1601-1609, Feb. 16, 1993.
Anal. Chem, (1993), vol. 55, pp. 1779-1784, "Enzyme Immunoassay Using a Rat Prolactin-Alkaline Phosphatase Recombinant Tracer", Daniel Gillet, et al.
Journal of Immunological Methods, vol. 169, 1994, pp. 205-211, "Recombinant Technology in the Preparation of Immunogen and Enzymatic Tracer. Application to the Development of an Enzyme Immunoassay for Rat Prolactin", Eric Ezan, et al.
Journal of Immunological Methods, vol. 181, 1995, pp. 177-186, "Recombinant Antibody-Alkaline Phosphatase Conjugates for Diagnosis of Human IgGs: Application to Anti-HBsAg Detection", Alice Carrier, et al.
Protein Engineering, vol. 5, No. 3, pp. 273-278, 1992, "Insertion of a Disulfide-Containing Neurotoxin into E. coli Alkaline Phosphatase: The Hybrid Retains both Biological Activities", Daniel Gillet, et al.
Bio/Technology, vol. 11, May 1993, pp. 601-605, Recombinant Colorimetric Antibodies: Construction and Characterization of a Bifunctional F(ab)2/Alkaline Phosphatase Conjugate Produced in Escherichia coli, Frederic Ducancel, et al.

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