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Expression and purification of recombinant human bone morphogenetic protein-7 (rhBMP-7) in Bacillus subtilis

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Abstract

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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Authors and Affiliations

  1. Department of Materials and Biochemical Engineering, Chonnam National University, Gwangju, 500-757, Korea

    Chun-Kwang Kim

  2. School of Applied Chemical Engineering, Chonnam National University, Gwangju, 500-757, Korea

    Jong Il Rhee

  3. Research Center for Biophotonics, Chonnam National University, Gwangju, 500-757, Korea

    Chun-Kwang Kim, Sung-Duk Oh & Jong Il Rhee

  4. Center for Functional Nano Fine Chemicals, Chonnam National University, Gwangju, 500-757, Korea

    Chun-Kwang Kim & Jong Il Rhee

  5. Department of Chemical & Biomolecular Engineering, North Carolina State University, Raleigh, NC, 27695-7905, USA

    Esther Meerim Lee

  6. Department of Orthopedic Surgery, Chonnam National University Hwasun Hospital, Jeonnam, 519-809, Korea

    Taek-Rim Yoon

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  1. Chun-Kwang Kim
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  2. Sung-Duk Oh
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  3. Jong Il Rhee
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  4. Esther Meerim Lee
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Correspondence to Jong Il Rhee.

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Kim, CK., Oh, SD., Rhee, J.I. et al. Expression and purification of recombinant human bone morphogenetic protein-7 (rhBMP-7) in Bacillus subtilis . Biotechnol Bioproc E 15, 830–836 (2010). https://doi.org/10.1007/s12257-009-3116-y

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  • DOI: https://doi.org/10.1007/s12257-009-3116-y

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