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Recombinant trypsin production in high cell density fed‐batch cultures in Escherichia coli
Author(s) -
Yee L.,
Blanch H. W.
Publication year - 1993
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260410804
Subject(s) - trypsin , recombinant dna , yield (engineering) , escherichia coli , periplasmic space , chemistry , protease , biochemistry , strain (injury) , chromatography , serine protease , enzyme , biology , materials science , anatomy , metallurgy , gene
Fed‐batch techniques were employed to obtain high cell density cultures (92–100 g DCW/L) of Escherichia coli strain X90 producing a recombinant serine protease, rat anionic trypsin, secreted to the periplasm. The specific growth rate was controlled to minimize growth‐inhibiting acetate formation by utilizing an exponential feeding profile determined from mass balance equation. The volumetric yield of recombinant rat anionic trypsin was 56 mg/L, and the final cell density was 92 g DCW/L when the culture was induced in the late logarithmic phase. However, when the culture was induced in the early logarithmic phase, the volumetric yield was 13 mg/L and the final cell density was 14 g DCW/L. Thus, the induction timing is shown to have a significant effect on the final cell density as well as the overall volumetric yield of the recombinant protease. © 1993 Wiley & Sons, Inc.