Secretory Production of Recombinant Protein by a High Cell Density Culture of a Protease Negative Mutant Escherichia coli Strain
- 5 April 1999
- journal article
- research article
- Published by Wiley in Biotechnology Progress
- Vol. 15 (2) , 164-167
- https://doi.org/10.1021/bp9900108
Abstract
Several protease negative mutant strains including HM114, HM126, and HM130 as well as their parent strain KS272 were compared for their growth and secretory production of a model fusion protein, protein A-beta-lactamase. HM114, a strain deficient in two cell envelope proteases, grew slightly faster and produced more fusion protein than the other strains deficient in more proteases. HM114 was grown to a cell dry weight of 47.86 g/L in 29 h using pH-stat, fed-batch cultivation. The beta-lactamase activity was 11.25 x 10(4) U/L, which was 30% higher than that obtained with its parent strain KS272. Up to 96% of protein A-beta-lactamase fusion protein could be recovered by a simple cold osmotic shock method. The specific beta-lactamase activity obtained with HM114 after fractionation was 4.5 times higher than that obtained with KS272.Keywords
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