Method for isolating an enzyme from a fermentation broth

Details Method for isolating an enzyme from a fermentation broth Method for isolating an enzyme from a fermentation broth

1991-08-27

1994-07-19

Naff, David M.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

435202, 435204, 435219, 435177, 435180, C12N 900, C12N 928, C12N 932, C12N 950

Patent

active

053309057

ABSTRACT:
To obtain enzymes from an enzyme-containing suspension, such as fermentation broth, the suspension is placed, together with particulate material capable of binding enzyme, such as ion exchange resin, in a vertical container. An ascending stream of inert gas, such as dry nitrogen, is passed through the resulting composition at a rate sufficient for fluidization. The resulting composition is mixed and evaporated until an adequate amount of enzyme is bound to the particulate material. Concentration of enzyme in the suspension is directly proportional to the amount of enzyme bound to particulate material. The residual suspension is forced out of the container by a pulse of compressed gas, and the bound enzyme is eluted. To speed up the process, the stream of gas and/or the composition are heated to enzyme-tolerated temperatures. The resulting composition can be further subjected to a washing step after removal of the residual suspension and prior to elution of the enzyme. The washing liquid can be passed through another fixed bed column containing particulate material capable of binding enzyme. Elution of the enzyme can be carried out as a stepwise process.

REFERENCES:
patent: 3943072 (1976-03-01), Thomson et al.
patent: 4318990 (1982-03-01), Thomson et al.
patent: 4690892 (1987-09-01), Ananthapadmanabhan et al.
Trevan, M. D. Immobilized Enzymes 1980, pp 68-71, John Wiley & Sons Ltd.
Shirai, Y. et al., Continuous production of monoclonal antibody with immobilized hybridoma cells in an expounded beci fermenter, Appli Microbiol Biotechnol 1987 26: pp.495-499.

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