*The Potential of Precision Probiotic* Hafnia alvei *HA4597 to Support Weight Loss DOI: http://dx.doi.org/10.5772/intechopen.103723*

**Figure 2.**

*From in vitro identification of ClpB to in vivo proof of concept.*

The functional mimetic properties of *E. coli* K12 ClpB were first confirmed with a proteomic approach. ClpB was identified after Western Blot analysis of *E. coli* K12 stationary phase total protein extract of *E. coli* K12 using anti alpha-MSH IgG [9]. Then, two pathways used by the ClpB to regulate satiety were identified with preliminary protocols in mice:


Anorexigenic properties of *E. coli* K12 ClpB were then confirmed *in vivo* on mice. Oral gavage of mice with *E. coli* K12 triggers a decrease of the food intake and body weight that wasn't shown with ClpB-deficient strains [14].

The alpha-MSH mimetic properties of bacterial *E. coli* ClpB made it a good candidate to develop an anti-obesity solution, but to develop a product with unquestionable safety, the team looked for a ClpB-producing strain with a food grade status. *Hafnia alvei,* a starter culture used in cheese production, in particular the Normandy traditional cheeses Camembert and Brie, was identified thanks to *in silico* approach [14]. *H. alvei* is a commensal bacterium from the *Enterobacteriaceae* family, naturally present in cheese [15, 16] and in the human gut [14], and thus benefitting from a long history of safe use.

The summary of these steps of discovery and development are exposed in **Figure 2** hereunder.
