[High density fed-batch culture of Escherichia coli DH5 alpha/pDH-B2m with DO feed-back control of nutrient feeding].
Optimization of cultivation condition of recombinant E. coli DH5 alpha/pDH-B2m and the condition suitable for expression of recombinant mature peptide of human bone morphogenetic protein-2 was carried out in 500 mL shaking flasks and then transferred to NBS Bioflo IV, a 20 L DO feed-back fed-batch culture system, to obtain rhBMP-2. The results indicate that keeping dissolved oxygen at 40% and controlling nutrient feeding rate with DO feed back strategy can obtain theoretically 3.59 g recombinant mature peptide of hBMP-2 per liter of broth, the final cell density OD600 reaches 57(22.8 g dry cell weight/L), and the expression of rhBMP-2 amounts to 30% of the total protein in E. coli.
Li
Yi
Y
Department of Biochemistry and Molecular Biology, Forth Military Medical University, Xi'an 710032, China.
Pu
Qin
Q
Zhao
Zhong-Liang
ZL
Chai
Yu-Bo
YB
Chen
Nan-Chun
NC
Chen
Su-Min
SM
D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis / Q000235:genetics*; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation; D005990:Glycerol / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013997:Time Factors; D016212:Transforming Growth Factor beta
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D003470:Culture Media; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; D005990:Glycerol