A new member of the bacterial ribonuclease inhibitor family from Saccharopolyspora erythraea
Open Access
- 19 December 2003
- journal article
- Published by Wiley in FEBS Letters
- Vol. 557 (1-3) , 164-168
- https://doi.org/10.1016/s0014-5793(03)01468-6
Abstract
We have identified Sti, the gene of a ribonuclease inhibitor from Saccharopolyspora erythraea, by using a T7 phage display system. A specific phage has been isolated from a genome library by a biopanning procedure, using RNase Sa3, a ribonuclease from Streptomyces aureofaciens, as bait. Sti, a protein of 121 amino acid residues, with molecular mass 13 059 Da, is a homolog of barstar and other microbial ribonuclease inhibitors. To overexpress its gene in Escherichia coli, we optimized the secondary structure of its mRNA by introducing a series of silent mutations. Soluble protein was isolated and purified to homogeneity. Inhibition constants of complex of Sti and RNase Sa3 or barnase were determined at pH 7 as 5×10−12 or 7×10−7, respectively.Keywords
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