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KMID : 1094720100150050830
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Biotechnology and Bioprocess Engineering
2010 Volume.15 No. 5 p.830 ~ p.836
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Expression and purification of recombinant human bone morphogenetic protein-7 (rhBMP-7) in Bacillus subtilis
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Kim Chun-Kwang
Oh Sung-Duk
Rhee Jong-Il
Lee Esther Mee-Rim
Yoon Taek-Rim
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Abstract
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The polypeptide representing the mature part of human bone morphogenetic protein-7 (BMP-7) was cloned and efficiently expressed in Bacillus subtilis. Recombinant B. subtilis had a clear band for rhBMP-7, a homodimeric protein with an apparent molecular weight of 15.4 kDa and produced 350 pg rhBMP-7/mL of culture medium. The extracellular and intracellular rhBMP-7 was purified in two steps using a fast performance liquid chromatography (FPLC) system with an ion-exchange column and a gel filtration column. The extracellular rhBMP-7 had a purity of 57.1% and a yield of 58.8%, while the purity of the intracellular rhBMP-7 was 36.2% with a yield of 51.4%. The rhBMP-7 produced in this work also stimulated alkaline phosphatase (ALP) activity in a dose-dependent manner, i.e. 2.5- and 8.9-fold at 100 and 300 ng rhBMP-7/mL, respectively, and showed intact biological activity.
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KEYWORD
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Bacillus subtilis, bone morphogenetic protein-7 (BMP-7), expression, purification
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