Molecular Analysis of Growth Inhibition Caused by Overexpression of the Biotin Operon in Escherichia coli

Abstract

Constitutive overexpression of the biotin operon (type 9 mutation) in a multicopy plasmid resulted in growth inhibition in Escherichia coli. Deletion analysis of the biotin operon indicated that overexpression of the bioB gene alone, the product of which is believed to catalyze the conversion of dethiobiotin to biotin, is sufficient for growth inhibition. This growth inhibition was still observed when the wild-type bioB gene was replaced by several mutant-type bioB genes derived from biotin auxotrophs that have base-pair substitutions creating amino acid substitutions in the bioB gene product. However, the modification of Ala 143 and Gly 99 of the bioB gene product resulted in recovery from growth inhibition. These results suggest that this phenotype of growth inhibition by overexpression of the bioB gene in E. coli is independent of the biotin-forming activity itself, but is caused by some function involving a specific conformation of the bioB gene product.