Refolding of porcine growth hormone from inclusion bodies of Escherichia coli

被引:11
作者
Baranauskaite, L [1 ]
Sereikaite, J [1 ]
Gedminiene, G [1 ]
Bumeliene, Z [1 ]
Bumelis, VA [1 ]
机构
[1] Vilnius Gediminas Tech Univ, Fac Fundamental Sci, Dept Chem & Bioengn, Vilnius, Lithuania
关键词
porcine growth hormone; inclusion bodies; refolding;
D O I
10.1080/10242420500175820
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Escherichia coli cells expressing porcine growth hormone were grown in a batch fermentation process. The expression level was estimated to be nearly 40% of the total cellular protein after 2-3 h of induction with 1 mM isopropyl beta-D-thiogalactoside. Porcine growth hormone expressed as inclusion bodies was solubilized in 8 M urea. Refolding conditions following a dilution protocol in the presence of beta-mercaptoethanol or using a glutathione pair were tested. Reverse phase-HPLC was applied to distinguish oxidized, misfolded and reduced forms of the hormone. A ratio of reduced to oxidized glutathione equal to 2/1 was chosen to avoid the formation of misfolded forms at high protein concentration.
引用
收藏
页码:185 / 189
页数:5
相关论文
共 16 条
[1]   EQUILIBRIUM DENATURATION OF RECOMBINANT PORCINE GROWTH-HORMONE [J].
BASTIRAS, S ;
WALLACE, JC .
BIOCHEMISTRY, 1992, 31 (38) :9304-9309
[2]  
BENTLE LA, 1987, Patent No. 4652630
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   COMPARING THE REFOLDING AND REOXIDATION OF RECOMBINANT PORCINE GROWTH-HORMONE FROM A UREA DENATURED STATE AND FROM ESCHERICHIA-COLI INCLUSION-BODIES [J].
CARDAMONE, M ;
PURI, NK ;
BRANDON, MR .
BIOCHEMISTRY, 1995, 34 (17) :5773-5794
[5]   Protein refolding for industrial processes [J].
Clark, ED .
CURRENT OPINION IN BIOTECHNOLOGY, 2001, 12 (02) :202-207
[6]  
Clark EDB, 1998, CURR OPIN BIOTECH, V9, P157
[7]  
FISHER B, 1993, BIOTECHNOL BIOENG, V41, P3
[8]   Estimating the instability parameters of plasmid-bearing cells. I. Chemostat culture [J].
Ganusov, VV ;
Brilkov, AV .
JOURNAL OF THEORETICAL BIOLOGY, 2002, 219 (02) :193-205
[9]  
Glick B. R., 2003, Molecular Biotechnology: Principles and Applications of Recombinant NA Technology, V3rd
[10]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+