High-level secretory production of recombinant human platelet-derived growth factor--BB by Saccharomyces cerevisiae under the non-selective conditions.
Recombinant human platelet-derived growth factor-BB (rhPDGF-BB) was produced by Saccharomyces cerevisiae. A two-stage cultivation strategy with mixture of glucose and galactose was developed to enhance rhPDGF-BB production, with its concentration being 32 mg/l fermentation broth under optimal conditions: corn steep powder as nitrogen source, 2 g/1 of glucose concentration at the beginning of induction phase, a pH of 5.0 and 6.5 for cell growth and rhPDGF-BB accumulation. The purification process consisted of yeast supernatant ultrafiltration followed by ion exchange chromatography and molecular sieve, and the recovery of rhPDGF-BB was estimated to be 20.28%. Biological activity of the purified rhPDGF-BB was 3.05 ng/ml.
Wang
Y
Y
College of Light Industry and Food Sciences, South China University of Technology, Guangzhou 510640, China. yonghw@scut.edu.cn
Xue
L
L
Li
Y
Y
Zhu
Y
Y
Yang
B
B
Wang
X
X
D000077214:Becaplermin; D006801:Humans; D010982:Platelet-Derived Growth Factor / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development*
D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; D000077214:Becaplermin