| US 7,455,990 B2 | ||
| Method of extracting recombinant hexose oxidase | ||
| Claus Lindvald Johansen, Højbjerg (Denmark); Søren Kjaerulff, Hillerød (Denmark); Susan Mampusta Madrid, Vedbaek (Denmark); Henrik Pedersen, Østbirk (Denmark); Charlotte Horsmans Poulsen, Braband (Denmark); and Masoud Rajabi Zargahi, Åbyhøj (Denmark) | ||
| Assigned to Danisco A/S, Copenhagen (Denmark) | ||
| Filed on Oct. 24, 2003, as Appl. No. 10/693,234. | ||
| Application 10/693234 is a continuation in part of application No. 09/722938, filed on Nov. 27, 2000, abandoned. | ||
| Application 09/722938 is a continuation of application No. PCT/IB00/01886, filed on Nov. 24, 2000. | ||
| Claims priority of application No. 9927801.2 (GB), filed on Nov. 24, 1999. | ||
| Prior Publication US 2004/0253671 A1, Dec. 16, 2004 | ||
| Int. Cl. C12P 21/06 (2006.01); C12N 9/04 (2006.01); C12N 1/21 (2006.01); C12N 1/15 (2006.01); C12N 1/19 (2006.01); C07H 21/04 (2006.01) | ||
| U.S. Cl. 435—69.1 [435/190; 435/252.3; 435/254.11; 536/23.2] | 13 Claims |
| 1. A method for extracting a soluble or membrane associated intracellular recombinant protein of interest (POI) from a bacterial,
yeast or fungal cell, the POI being released from the bacterial, yeast or fungal cell, comprising the steps of:
(a) providing a bacterial, yeast or fungal cell comprising a soluble or membrane associated intracellular recombinant POI;
(b) releasing the recombinant POI from the cell by contacting the cell with a membrane extracting composition comprising a
quaternary ammonium compound at a concentration of between 0.05% to 0.6% by weight, under conditions sufficient for the release
of the recombinant POI in a soluble form; and,
(c) recovering the recombinant POI from the membrane extracting composition; wherein the quaternary ammonium compound is selected from the group consisting of Lauroyl Trimethyl Ammonium Bromide (LTAB),
Myristyl Trimethyl Ammonium Chloride (MTAC), CetylTrimethyl Ammonium Chloride (CTAC), Cetrimide, Cetyl Trimethyl Ammonium
Bromide (CTAB), Stearoyl Trimethyl Ammonium Chloride (STAC), Stearoyl Trimethyl Ammonium Bromide (STAB), Benzalkonium Chloride
(alkyldimethylbenzylammonium chloride), N-Cetylpyridinium Bromide (N-Hexadecylpyridinium bromide), N-Cetylpyridinium Chloride
(N-Hexadecylpyridinium chloride), Benzyl Dimethyl Tetradecyl Ammonium Chloride, Benzyl Dimethyl Hexadecyl Ammonium Chloride
and a combination of any two or more thereof, and wherein the recombinant POI is a hexose oxidase (HOX) enzyme.
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