[Expression of recombinant human serum albumin in genetically engineered Pichia pastoris in high-density fermentation].
The optimum culture conditions of genetically engineered Pichia pastoris in shake-flask cultivation and in fed-batch fermentation were investigated respectively in this paper. It showed that the cultural period induced with 5 g/L methanol is 96 h, optimum methanol concentration is 10 g/L, and pH range is 5.72-6.59 in shake-flask culture. Although the seed inoculum amount increased, the target protein productivity per cell optical density was decreased. Their relationship fit the equeation Y = 12.941x(-0.5059) (r = 0.9789, where x is inoculating OD600, Y is protein productivity per cell optical density), we postulated that the restricting factor may be dissolved oxygen (DO) at shake-flask culture. With the 10% inoculum and 20 OD600 of seed, the lag phase of cell growth is 2.11 h in batch cultivation, and the relationship between cell optical density (Y) and culture time (t) is Y = 0.7841e0.2319t (r = 0.9936); The cell dry weight of broth reached 115-160 g/L and the maximum rHSA concentration was 3.6 g/L at the 120th at the fed-batch fermentation phase.
Guo
Meijin
M
State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237, China.
Zhuang
Yingping
Y
Chu
Ju
J
Zhang
Siliang
S
Liu
Zhimin
Z
D005285:Fermentation; D005818:Genetic Engineering; D010843:Pichia / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012709:Serum Albumin / Q000096:biosynthesis*
D011994:Recombinant Proteins; D012709:Serum Albumin