The solubility of recombinant proteins expressed in Escherichia coli is increased by otsA and otsB co-transformation.
The osmolyte trehalose strongly limits protein aggregation both in vitro and in vivo. The addition of trehalose to the culture medium reduced the aggregation of recombinant proteins expressed in Escherichia coli in a concentration-dependent manner. Comparable positive effects were obtained when the host bacteria were engineered to overexpress the gene products of otsA and otsB, the two enzymes involved in trehalose synthesis. Apparently, the osmolyte preserves protein monodispersion rather than directly facilitating protein folding. However, the stabilization of the protein folding intermediate(s) resulted in higher yields of native proteins and aggregates of lower complexity. Other osmolytes have been tested in vitro in comparison with trehalose. Di-myo-inositol1,1'-phosphate (DIP) seems to be a good candidate to test in in vivo applications, although the opportunity of using otsA/B overexpressing cells is simpler and less expensive.
Schultz
Tina
T
EMBL Scientific Core Facilities, Meyerhofstr. 1, D-69117 Heidelberg, Germany.
Liu
Jing
J
Capasso
Paola
P
de Marco
Ario
A
D000599:Amino Acids, Diamino / Q000494:pharmacology; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism*; D005822:Genetic Vectors; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism; D012995:Solubility; D014170:Transformation, Genetic; D014199:Trehalose / Q000494:pharmacology
D000599:Amino Acids, Diamino; D017931:DNA Primers; D018076:DNA, Complementary; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C121557:hydroxyectoine; D014199:Trehalose; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase; C518654:OtsB protein, E coli; D010744:Phosphoric Monoester Hydrolases