Expression of recombinant proteinase K from Tritirachium...

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase

Reexamination Certificate

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C435S320100, C435S254230, C536S023200

Reexamination Certificate

active

10468252

ABSTRACT:
The invention concerns a method for the expression of a gene coding for a soluble proteinase K in yeast e.g. inPichia pastoriswith subsequent secretion into the culture medium. In addition a method for purifying the heterologously expressed and secreted proteinase K is described.

REFERENCES:
patent: 0396106 (1990-11-01), None
patent: WO 87/02673 (1987-05-01), None
patent: WO 88/07581 (1988-10-01), None
patent: WO 96/28556 (1996-09-01), None
1998 INVITROGEN Catalog “The Pichia Expression System” pp. 19-23.
Clark, Elaina De Bernardez et al., “Inhibition of Aggregation Side Reactions during in Vitro Protein Folding,” Methods in Enzymology, vol. 309, pp. 217-236 (1999).
Ebeling, Wolfgang et al., “ProteinaseK fromTritirachium albumLimber,” Eur. J. Biochem., 47, 91-97 (1974).
Gunkel, F. Andreas et al., “Proteinase K fromTritirachium albumLimber,” Eur. J. Biochem. 179-185-194 (1989).
Kane, James F. et al., “Effects of rare codon clusters on high-level expression of heterologous proteins inEscherichia coli,” Current Opinion in Biotechnology 1995, 6:494-500.
Penheiter, Alan R. et al., “Purification and Characterization of a Soybean Root Nodule Phosphatase Expressed inPichia pastoris,” Protein Expression and Purification 14, 125-130 (1998).
Samal, Babru et al., “Cloning and Expression of the Gene Encoding a Novel Proteinase fromTritirachium albumLimber,” Practical Protein Engineering, 1996, pp. 95-104.
Cregg, J. et al., “Recombinant Protein Expression inPichia pastoris,” Mol. Biotechnol. vol. 16, No. 1, (2000). 23-52.
Roy, N. et al., “Expression of Human Gelatinase B inPichia pastoris, ” Protein Expression and Purification, 16(1999) 324-330.
Borg-von Zepelin, M. et al., “The Expression of the Secreted Aspartyl Proteinases Sap4 to Sap6 fromCandida albicansin Murine Macrophages,” Mol. Microbiol. vol. 28, No. 3, (1998) 543-554.
Hiroshi Nakakubo, et al., “Secretory Production of Recombinant Human Chymase as an Active Form inPichia pastoris, ” Yeast 16(2000) 315-323.
Withers-Martinez, C. et al., “PCR-based Gene Synthesis As An Efficient Approach For Expression Of The A + T-rich Malaria Genome,” Protein Eng. vol. 12, No. 12(1999), 1113-1120.
Hiroshi Nakakubo et al., “Functional Reconstitution of an Active Recombinant Human Chymase FromPichia pastorisCell Lysate,” Yeast 16 (2000), 1387-1396.

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