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

被引：6

作者：

Kim, Chun-Kwang ^{[1
]}

Oh, Sung-Duk

Rhee, Jong Il ^{[1
]}

Lee, Esther Meerim ^{[2
]}

Yoon, Taek-Rim ^{[3
]}

机构：

[1] Chonnam Natl Univ, Ctr Funct Nano Fine Chem, Kwangju 500757, South Korea

[2] N Carolina State Univ, Dept Chem & Biomol Engn, Raleigh, NC 27695 USA

[3] Chonnam Natl Univ, Hwasun Hosp, Dept Orthoped Surg, Jeonnam 519809, South Korea

来源：

BIOTECHNOLOGY AND BIOPROCESS ENGINEERING | 2010年 / 15卷 / 05期

关键词：

Bacillus subtilis; bone morphogenetic protein-7 (BMP-7); expression; purification; GROWTH-FACTOR-BETA; ESCHERICHIA-COLI; SECRETION; SYSTEM;

D O I：

10.1007/s12257-009-3116-y

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页码：830 / 836

页数：7

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