Production of human recombinant bone morphogenetic protein-2A by high density culture of Escherichia coli with stationary dissolved oxygen fed-batch condition.
The optimization of cultivation condition in 500 ml shake flasks was carried out to produce recombinant human bone morphogenetic protein-2A (BMP-2A) in recombinant Escherichia coli YK537/pDH-B2m, followed by a 5L fermenter batch and condition-controlled fed-batch culture to obtain BMP-2A. The comparison of these two methods indicated that cultivation by keeping dissolved oxygen at 30%-40% and limiting glucose concentration could obtain BMP-2A 2.78 g/L broth, the final cell density was OD600 53 (dry cell weight 21.2 g/L), and expressed BMP-2A was 25% of the total amount of protein in E.coli. The critical fermentation conditions included: (1) keeping appropriate dissolved oxygen concentration in the process; (2) limiting glucose concentration; (3) taking heat induction at the middle-log phase and maintaining 42 degrees C for 4 hours; (4) controlling specific growth rate around 0.3 h-1 in the duration of growth.
Li
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M
Shanghai Research Center of Biotechnology, Chinese Academy of Sciences, China.
Chen
C
C
Pu
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Q
Chen
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S
D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D010100:Oxygen / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; D010100:Oxygen