Pilot scale production of a recombinant human epidermal growth factor, secreted by Bacillus brevis, using expanded bed adsorption |
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Authors: | A Miyauchi S Ebisu K Uchida M Yoshida M Ozawa T Tojo K Kadowaki H Takagi |
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Institution: | (1) Research Laboratory, Higeta Shoyu Co, 2–8 Chuo-cho, Choshi-shi, Chiba 288–8680, Japan, JP |
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Abstract: | Recombinant Bacillus brevis which carried an expression plasmid encoding the human epidermal growth factor (EGF) gene on a cryptic high-copy number plasmid,
pHT926, extracellularly produced EGF in its biologically active form at a concentration of over 1.5 g L−1 in the culture broth in a 30-L jar fermenter. The culture broth also contained some other EGF compounds, which mainly consisted
of oligomeric and polymeric forms with disulfide bonds. We developed a simple purification method for EGF, without prior cell
removal from the culture broth, comprising cation exchange expanded bed adsorption followed by ultrafiltration with UF 10 000
and 3000 membranes. The EGF compounds were efficiently separated from the EGF in its native form in the expanded bed adsorption
step. With this purification method, only EGF in its native form was recovered from the culture broth, with a yield of nearly
80%, and 90% purity. This efficient and economic system has made it possible to use EGF as a pharmaceutical in the livestock
industry.
Received 10 June 1998/ Accepted in revised form 10 September 1998 |
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Keywords: | : human epidermal growth factor Bacillus brevis recombinants expanded bed adsorption fermentation |
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