High cell density cultivation of recombinant Escherichia coli for hirudin variant 1 production.
A synthetic medium, TK-25, for high cell density cultivation (HCDC) of Escherichia coli K-12 was modified to support HCDC of strain JM109. By optimizing the culture conditions, the cell concentration of 65 g/l in 14 h was obtained in the optimized medium, namely TK-10, with glucose-fed batch cultivation. When these conditions were further applied for HCDC of E. coli JM109 harboring pUC-based recombinant plasmid which expresses a hirudin variant, HV-1-fused protein under the control of trp promoter, it grew to 24 g/l of dried cells expressed as an inclusion body as 15.9% of the total protein, corresponding to 1908 mg/l hirudin-fused protein.
Matsui
Toru
T
Center of Molecular Biosciences, University of the Ryukyus, 1 Sembaru Nishihara-cho, Okinawa 903-0213, Japan. tmatsui@comb.u-ryukyu.ac.jp
Sato
Hiroaki
H
Yamamuro
Hotaka
H
Misawa
Satoru
S
Shinzato
Naoya
N
Matsuda
Hitoshi
H
Takahashi
Joji
J
Sato
Seigo
S
D002455:Cell Division; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics*; D006629:Hirudins / Q000235:genetics* / Q000378:metabolism; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism
D006629:Hirudins; D011994:Recombinant Proteins