Crossflow microfiltration process for the separation of biotechnologically produced materials

• US 4,751,003 A
• Filed: 05/01/1986
• Issued: 06/14/1988
• Est. Priority Date: 05/02/1985
• Status: Expired due to Fees

• Current Assignee: Henkel KGaA (Henkel AG & Co KGaA), Biochemie GmbH (Novartis Ag)
• Original Assignee: Henkel KGaA (Henkel AG & Co KGaA)
• Inventors: Raehse, Wilfried, Carduck, Franz-Josef, Kuehne, Norbert
• Primary Examiner(s): Castel, Benoit
• Application Number:
  US06/858,408
• Time in Patent Office: 775 Days
• Field of Search: 210/651, 210/641, 210/650, 210/638, 210/639, 210/500.41, 210/653, 210/805, 210/806, 435/803, 435/212, 435/261, 435/219-222, 435/813-816
• US Class Current: 210/639
• CPC Class Codes: B01D 2315/10 Cross-flow filtration
• CPC Class Codes: B01D 61/147 Microfiltration
• CPC Class Codes: B01D 63/06 Tubular membrane modules
• CPC Class Codes: C12M 29/02 Percolation
• CPC Class Codes: C12M 33/14 with filters, sieves or membranes
• CPC Class Codes: C12M 47/10 Separation or concentration of fermentation products bioreactors combined with means for distillation or extraction of liquid fuel C12M43/02
• CPC Class Codes: C12N 9/50 Proteinases , e.g. Endopeptidases (3.4.21-3.4.25)
• CPC Class Codes: C12N 9/52 derived from bacteria or Archaea
• CPC Class Codes: C12P 1/00 Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
• CPC Class Codes: Y10S 435/814 Enzyme separation or purification

Abstract

The invention relates to a process for the separation of biotechnologically produced valuable materials form a cell suspension by crossflow microfiltration to obtain an high specific permeate flux while retention stays near 0% for long periods. To enable crossflow microfiltration to be carried out on an industrial scale in biotechnology in the separation of biotechnologically produced extracellular valuable materials from a cell suspension, particularly in the separation of alkaline protease for recovering enzyme, under economically acceptacle conditions, alkaline protease of relatively high molecular weight, more especially enzyme >20,000 daltons, is separated from a fermenter solution using polysulfone tubes having micropore diameters of from 0.3 to 0.5 μm at a rate of flow of the fermenter solution of from 3 to 6 m/s parallel to the membrane surface and with a pressure difference between the concentrate side and the permeate side of 2 bar, the ratio of the mean pore diameter of the membrane to the size of the microorganisms remaining in the concentrate being between 0.15 and 0.85

Claims

1. A crossflow microfiltration process for the separation of biotechnologically produced materials from a cell suspension comprising

(A) passing a cell suspension containing biotechnologically produced materials through a tubular, polysulfone, polymeric, microfiltration membrane having an inner surface and an outer surface, in crossflow to said inner surface, wherein

(a) said cell suspension flows through the tubular microfiltration membrane at about 3 to 6 meters per second,

(b) said microfiltration membrane has micropores with a mean diameter of about 0.3 to 0.5 μ

m,

(c) said mircopores are about 0.15 to 0.85 times the size of cells in said cell suspension, and

(d) there is a mean pressure gradient of about 2 bar between the inner surface of said microfiltration membrane and its outer surface,

so that the materials to be separated pass through said microfiltration membrane as a filtrate and the cell suspension remaining becomes a concentrate; and

(B) recovering said filtrate.

2. The process of claim 1 wherein solids having a particle size of not more than 500 μ

m are added to said cell suspension in an amount effective to increase the flux of filtrate through said microfiltration membrane.

3. The process of claim 2 wherein about 0.05 to 1.0% by weight of solids are added, based upon the weight of said cell suspension.

4. The process of claim 3 wherein said solids are ground cellulose.

5. The process of claim 1 wherein said permeate is an alkaline protease.

6. The process of claim 1 wherein the pH of said cell suspension is adjusted to about 6.2 to 7.2 before microfiltration.

7. The process of claim 5 wherein a surfactant is added to said cell suspension before microfiltration.

8. The process of claim 1 wherein said permeate is an alkaline protease enzyme of more than 20,000 daltons.

9. The process of claim 1 further comprising serially passing said cell suspension through a plurality of the microfiltration membranes which are serially connected, and recycling cell suspension concentrate leaving the last of said plurality to the first of said plurality.

10. The process of claim 9 further comprising enriching the recovered filtrate by subjecting it to ultrafiltration.

11. The process of claim 1 further comprising enriching the recovered filtrate by subjecting it to ultrafiltration.