ISSN:
1476-5535
Keywords:
Keywords: human epidermal growth factor; Bacillus brevis recombinants; expanded bed adsorption; fermentation
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Process Engineering, Biotechnology, Nutrition Technology
Notes:
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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1038/sj.jim.2900570
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