Journal
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
Volume 165, Issue 2, Pages 416-425Publisher
SPRINGER
DOI: 10.1007/s12010-011-9261-8
Keywords
2-Deoxyribose-5-phosphate aldolase; Lactose induction; Continuous; Glucose limited; Fed-batch cultivation
Funding
- Major Science and Technology Project of Zhejiang Province [2007 C01004-2]
- National Natural Science Foundation of China [20906016]
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The production of a thermophilic 2-deoxyribose-5-phosphate aldolases (DERA) in Escherichia coli BL21 under continuous lactose induction strategy was investigated. The process was combined with the exponential feeding method, controlling the feeding rate to maintain the specific growth rate at 0.15 h(-1). The results indicate that the lactose concentration in the feed medium affected directly the expression of the target protein. The use of 50 g/L in the feed medium resulted in the biomass concentration of 39.3 g DCW/L, and an expression level of above 30%, and the maximum final DERA concentration of 16,200 U/L. Furthermore, the acetate concentration remained at a low level in the fed-batch phase, less than 0.5 g/L. In conclusion, combining glucose feeding with lactose induction is a more powerful way to achieve high cell density cultures and to efficiently produce the thermophilic DERA. The results also indicate the potential industrial utility in the scale production of other recombinant proteins.
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