Expression and Purification of Recombinant Human Bone Morphogenetic Protein-7 (rhBMP-7) in Bacillus subtilis

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/...

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Published in:Biotechnology and bioprocess engineering 2010, 15(5), , pp.830-836
Main Authors: Kim, C.K., Chonnam National University, Gwangju, Republic of Korea, Oh, S.D., Chonnam National University, Gwangju, Republic of Korea, Rhee, J.I., Chonnam National University, Gwangju, Republic of Korea, Lee, Esther Meerim, North Carolina State University, Raleigh, NC, USA, Yoon, T.R., Chonnam National University Hwasun Hospital, Hwasun, Republic of Korea
Format: Article
Language:English
Subjects:
Alkaline phosphatase
BACILLUS SUBTILIS
Biological activity
Biotechnology
Bone morphogenetic protein 7
Bone surgery
Bones
Cell culture
Chemistry
Chemistry and Materials Science
Chromatography
Cloning
E coli
Engineering
Enzymes
expression
Filtration
Gene expression
Industrial and Production Engineering
Liquid chromatography
Molecular weight
Phosphatase
Plasmids
Polypeptides
protein purification
Proteins
PURIFICACION
PURIFICATION
R&D
Research & development
Research Paper
Studies
생물공학
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Summary: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.
ISSN:1226-8372
1976-3816
DOI:10.1007/s12257-009-3116-y