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High-level production of Fc-fused kringle domain in Pichia pastoris
- Jeong, Gu Min, Lee, Yong Jae, Kim, Yong Sung, Jeong, Ki Jun
- Journal of industrial microbiology & biotechnology 2014 v.41 no.6 pp. 989-996
- Pichia pastoris, agonists, antagonists, bioreactors, carbon, glycerol, humans, pH, temperature
- Recently, as a new non-immunoglobulin-based protein scaffold, a human kringle domain was successfully engineered toward biologically functional agonists and antagonists. In this study, the fed-batch cultivation conditions were optimized for enhanced production of an Fc-fused kringle domain (KD548-Fc) in Pichia pastoris. Fed-batch cultivations were performed in 5-l laboratory-scale bioreactors, and in order to find the optimal conditions for high-level production of KD548-Fc, several parameters including the initial carbon source (glycerol) concentration, temperature, and pH were investigated. When cells were cultivated at pH 4.0 and 25 °C with 9.5 % glycerol in the initial medium, the highest production yield (635 mg/l) was achieved with high productivity (7.2 mg/l/h). Furthermore, functional KD548-Fc was successfully purified from the culture broth using a simple purification procedure with high purity and recovery yield.