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Claims for Patent: 6,204,012

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Summary for Patent: 6,204,012
Title: Protein production process
Abstract:A method for producing a protein by culturing a cell capable of expressing the protein and which contains a methylotrophic yeast promoter for an enzyme of the methanol metabolic pathway controlling expression of the protein, in a fermentative batch process comprising a batch phase and a feeding phase, under conditions such that dissolved oxygen is continually present in the culture medium throughout the process, and about 1% to about 100% of the carbon source during the feeding phase is a sugar or sugar polymer, which sugar or sugar polymer is provided in such an amount that the sugar or sugar polymer is continually depleted by the cell and therefore is substantially undetectable in the culture medium; and isolating the protein on completion of the feeding phase.
Inventor(s): Hellmuth; Karsten (Marklohe, DE), Lopez-Ulibarri; Rual (Montclair, NJ), Mayer; Anne Fran.cedilla.oise (New York, NY), Schlieker; Heinrich Winfried (Bloomfield, NJ), van Loon; Adolphus (Rheinfelden, CH)
Assignee: Roche Vitamins Inc. (Parsippany, NJ)
Application Number:09/163,052
Patent Claims:1. A method for producing a desired protein which comprises culturing a cell capable of expressing the protein comprising a nucleic acid sequence encoding the desired protein operably linked to a methylotrophic yeast promoter having an activity for an enzyme of the methanol metabolic pathway and controlling expression of the desired protein, in a fermentative batch process comprising a batch phase and a feeding phase under conditions such that dissolved oxygen is continually present in the culture medium throughout the process, and about 1% to about 100% of the total carbon source present in the feeding phase is a repressive sugar or a repressive sugar polymer, which is provided in such an amount that the repressive sugar or repressive sugar polymer is continually depleted by the cell and therefore substantially undetectable in the culture medium; and isolating the protein on completion of the feeding phase by collecting and purifying the desired protein from the culture medium.

2. A method of claim 1 wherein the protein is an enzyme.

3. A method of claim 2 wherein the protein is phytase, cellulase, xylanase, amylase, protease, invertase, lipase, catalase, cellulase, glucose oxidase, alcohol oxidase, pectinase, naraginase, collagenase, peroxidase or pullulanase.

4. A method of claim 1 wherein the cell is a methylotrophic yeast cell.

5. A method of claim 4 wherein the yeast cell is a Hansenula, Pichia, Candida or Torulopsis cell.

6. A method of claim 5 wherein the yeast cell is Hansenula polymorpha or Pichia pastoris.

7. A method of claim 1 wherein the promoter is the formate dehydrogenase promoter, the methanol oxidase promoter or the dihydroxyacetone synthase promoter.

8. A method of claim 1 wherein the cell is eukaryotic host cell which has been engineered to contain the methylotrophic yeast promoter.

9. A method of claim 7 wherein the cell is eukaryotic host cell which has been engineered to contain the methylotrophic yeast promoter.

10. A method of claim 1 wherein the sugar or sugar polymer is a mono-, di-, oligo- or polysaccharide.

11. A method of claim 10 wherein the sugar or sugar polymer is glucose, fructose, sucrose, maltose, starch, glycogen, cellulose or dextrose.

12. A method of claim 11 wherein the sugar is glucose.

13. A method of claim 1 wherein the sugar is in a sugar containing composition which is a natural or artificially produced syrup.

14. A method of claim 13 wherein the sugar containing composition is molasses, glucose syrup, or fructose syrup.

15. A method of claim 14 wherein the sugar containing composition is glucose syrup.

16. A method of claim 1 wherein the sugar or sugar polymer makes up about 40% to about 100% of the carbon source.

17. A method of claim 16 wherein the sugar or sugar polymer makes up about 90% to about 100% of the carbon source.

18. A method of claim 17 wherein the sugar or sugar polymer makes up about 100% of the carbon source.

19. A method of claim 18 wherein the sugar or sugar polymer is glucose.

20. A method of claim 1 wherein the dissolved oxygen is present in the culture medium at a level of from about 1% to about 100% saturation.

21. A method of claim 20 wherein the dissolved oxygen is present in the culture medium at a level of from about 10% to about 100% saturation.

22. A method of claim 21 wherein the dissolved oxygen is present in the culture medium at a level of about 20% saturation.

23. A method of claim 19 wherein the dissolved oxygen is present in the culture medium at a level of about 20% saturation.

Summary for Patent:   Start Trial

Foriegn Application Priority Data
Foreign Country Foreign Patent Number Foreign Patent Date
97117021Oct 01, 1997

Details for Patent 6,204,012

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Smith And Nephew SANTYL collagenase OINTMENT;TOPICAL 101995 001 1965-06-04   Start Trial Roche Vitamins Inc. (Parsippany, NJ) 2017-10-01 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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