Continuous fermentation process

Abstract

A fermentation assembly comprising: (a) a vessel for culturing living cells; (b) at least two storage flasks in fluid communication with the vessel for supply of liquids and a first transport means for transferring the liquids from the storage flasks to the vessel; (c) individual appliances operably connected to the transport means for monitoring the supply of the contents of the storage flasks to the vessel; (d) a harvest flask in fluid communication with the vessel and a second transport means for transferring the fermentation broth from the vessel to the harvest flask; and (e) a device operably connected to the first transport means for controlling and maintaining a constant dilution rate in the vessel with varying rates of individual supply of liquid from the storage flasks to the vessel is disclosed.

What is claimed is:  
     
         1 . A fermentation assembly comprising 
 (a) a vessel for culturing living cells;    (b) at least two storage flasks in fluid communication with the vessel for supply of liquids and a first transport means for transferring the liquids from the storage flasks to the vessel;    (c) individual appliances operably connected to the transport means for monitoring the supply of the contents of the storage flasks to the vessel;    (d) a harvest flask in fluid communication with the vessel and a second transport means for transferring the fermentation broth from the vessel to the harvest flask; and    (e) a device operably connected to the first transport means for controlling and maintaining a constant dilution rate in the vessel with varying rates of individual supply of liquid from the storage flasks to the vessel.    
     
     
         2 . A fermentation assembly according to  claim 1  wherein the at least two storage flasks comprise individual storage flasks for solutions of carbon, nitrogen, and mineral sources.  
     
     
         3 . A fermentation assembly according to  claim 1  wherein at least one of the at least two storage flasks contains a controlling agent.  
     
     
         4 . A fermentation assembly according to  claim 1  further comprising an additional storage flask containing water.  
     
     
         5 . A fermentation assembly according to  claim 1  wherein the vessel contains a bed of immobilized living cells.  
     
     
         6 . A fermentation assembly according to,  claim 5  wherein the bed of immobilized cells is selected from the group consisting of a fixed bed, an expanded bed, a moving bed, and combinations thereof.  
     
     
         7 . An assembly according to  claim 5  wherein the living cells are immobilized on a porous carrier.  
     
     
         8 . A process for the manufacture of a protein comprising: 
 (a) providing a continuous culture of living cells in a fermentation reactor; and    (b) individually feeding nutrients and other agents required for the growth of the cells into the reactor at a constant dilution rate to achieve optimal production of the protein.    
     
     
         9 . A process according to  claim 8  wherein the protein is selected from the group consisting of catalase, lactase, phenoloxidase, oxidase, oxidoreductase, glucanase, cellulase, xylanase and other polysaccharides peroxidase, lipase, hydrolase, esterase, cutinase, protease and other proteolytic enzymes, aminopeptidase, carboxypeptidase, phytase, lyase, pectinase, pectinolytic enzymes, amylase, glucosidase, mannosidase, isomerase, invertase, transferase, ribonuclease, chitinase, and desoxyribonuclease.  
     
     
         10 . A process according to  claim 8  wherein the protein is a therapeutic protein.  
     
     
         11 . A process according to  claim 10  wherein the therapeutic protein is selected from the group consisting of antibodies, vaccines, and antigens.  
     
     
         12 . A process according to  claim 8  wherein the protein is an antibacterial and/or health-beneficial protein.  
     
     
         13 . A process according to  claim 12  wherein the antibacterial and/or health-beneficial protein is selected from the group consisting of lactoternin, lactoperoxidase and lysozyme.  
     
     
         14 . A process according to  claim 8  wherein the cells are immobilized.  
     
     
         15 . A process according to  claim 8  wherein the cell is a phytase-producing microorganism.  
     
     
         16 . A process according to  claim 15  wherein the phytase-producing microorganism is  Hansenula polymorpha.    
     
     
         17 . A process according to  claim 16  wherein the phytase-producing microorganism is  Hansenula polymorpha  transformed by a DNA encoding a phytase of fungal or consensus origin.  
     
     
         18 . A process according to  claim 8  wherein the fermentation reactor contains a bed of immobilized cells on a porous carrier.  
     
     
         19 . A process according to  claim 18  wherein the bed is selected from the group consisting of a fixed bed, an expanded bed, a moving bed, and combinations thereof.  
     
     
         20 . A process according to  claim 8  wherein the nutrients comprises a carbon source.  
     
     
         21 . A process according to  claim 20  wherein the carbon source is glycerol or sugar.  
     
     
         22 . A process according to  claim 21  wherein the sugar is selected from the group consisting of mono-, di-, and polysaccharides.  
     
     
         23 . A process according to  claim 20  wherein t he carbon source is glucose.  
     
     
         24 . A process according to  claim 20  wherein the carbon source is methanol.  
     
     
         25 . A process according to  claim 20  wherein the carbon source is glucose and methanol.  
     
     
         26 . A process according to  claim 25  wherein the total amount of methanol and glucose is from about 10 g/l to about 500 g/l each.  
     
     
         27 . A fermentation assembly comprising: 
 (a) a fermentor  1  equipped with inlet tubes  2   a  in fluid communication with a storage flask  2  for supply of liquids to the fermentor;    (b) a pump  3  operably connected to the inlet tubes for transporting liquids from the storage flask  2  to the fermentor  1 ;    (c) a scale  4  in contact with each storage flask for monitoring the amount of liquid supplied to and discharged from the fermentor;    (d) a gas inlet  9  and out let tubes  10  in communication with the fermentor for introducing and removing gas therefrom;    (e) a pump  6  operably connected to an outlet tube  5   a  which is in fluid connection with the fermentor, wherein the pump discharges fermentation broth from the fermentor to a harvest flask  5 ;    (f) a main controlling unit  7  operably connected to the fermentation assembly for overall process monitoring and steering;    (g) a controlling unit  11  operably connected to individual control systems  17  for monitoring and steering temperature, pH, gas pressure, fermentor content, and antifoam agents;    (h) a circuit  12  for monitoring gas supply and taking samples including an outlet tube from the fermentor, which circuit is operably connected to the outlet to be pump  13 ; and    (i) gas inlet and outlet flow control devices  14  and  15  operably connected to the gas inlet and outlet tubes  9 ,  10 .    
     
     
         29 . A fermentation assembly according to claim  28  further comprising sterile filters  16  and thermostating unit  8 .