The Bacillus licheniformis BlaP beta-lactamase as a model protein scaffold to study the insertion of protein fragments

Protein Sci. 2007 Oct;16(10):2260-71. doi: 10.1110/ps.072912407.

Abstract

Using genetic engineering technologies, the chitin-binding domain (ChBD) of the human macrophage chitotriosidase has been inserted into the host protein BlaP, a class A beta-lactamase produced by Bacillus licheniformis. The product of this construction behaved as a soluble chimeric protein that conserves both the capacity to bind chitin and to hydrolyze beta-lactam moiety. Here we describe the biochemical and biophysical properties of this protein (BlaPChBD). This work contributes to a better understanding of the reciprocal structural and functional effects of the insertion on the host protein scaffold and the heterologous structured protein fragments. The use of BlaP as a protein carrier represents an efficient approach to the functional study of heterologous protein fragments.

MeSH terms

  • Bacillus / enzymology*
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Chitin / metabolism
  • Enzyme Stability
  • Hexosaminidases / chemistry
  • Models, Molecular
  • Protein Denaturation
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / chemistry*
  • beta-Lactamases / chemistry*
  • beta-Lactamases / genetics

Substances

  • Bacterial Proteins
  • Recombinant Fusion Proteins
  • Chitin
  • Hexosaminidases
  • chitotriosidase
  • beta-Lactamases