Reduction of growth and acetyl-CoA carboxylase activity by expression of a chimeric streptavidin gene in Escherichia coli.
The streptavidin gene from Streptomyces avidinii was expressed in E. coli as a non-fusion protein and as a glutathione S-transferase fusion protein. The streptavidin protein accumulated primarily in the inclusion bodies and did not alter cell growth. In contrast, the glutathione-S-transferase-streptavidin fusion protein was soluble. Nondenaturing polyacrylamide gel electrophoresis indicated that the chimeric glutathione-S-transferase-streptavidin protein was present mostly as a monomer, with some detectable polymeric forms. Cells grown in the presence of [3H]-biotin had label specifically associated with the expressed glutathione-S-transferase-streptavidin fusion protein, indicating this protein bound biotin in vivo. The majority of the radiolabeled biotin was associated with polymeric forms of the glutathione-S-transferase-streptavidin protein. The growth rates of biotin auxotrophs of E. coli growing in biotin-deficient media were substantially decreased by the expression of the glutathione-S-transferase-streptavidin gene. The decreased growth rate correlated with a decrease in acetyl-CoA carboxylase activity.
Guan
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Department of Botany, Iowa State University, Ames 50011, USA.
Wurtele
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D000103:Acetyl-CoA Carboxylase / Q000096:biosynthesis / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001710:Biotin / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D015870:Gene Expression; D005798:Genes, Bacterial / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D008970:Molecular Weight; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D012995:Solubility; D019809:Streptavidin; D013302:Streptomyces / Q000737:chemistry
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D001710:Biotin; D019809:Streptavidin; D005982:Glutathione Transferase; D000103:Acetyl-CoA Carboxylase