Part:BBa_K5283019

P9, an 84 kDa thermogenesis-inducing protein secreted by Akkermansia muciniphila, enhances GLP-1 sec

The P9 protein gene from Akkermansia muciniphila is a bioengineered component with significant implications for the treatment of metabolic disorders such as diabetes and obesity. This gene encodes for an 84 kDa protein that has been shown to stimulate the secretion of glucagon-like peptide-1 (GLP- 1), a key regulator of energy metabolism and glucose homeostasis. The therapeutic potential of P9 lies in its ability to interact with intercellular adhesion molecule 2 (ICAM-2), which is crucial for its metabolic effects.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 43
  Illegal PstI site found at 1372
  Illegal PstI site found at 1744
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 43
  Illegal PstI site found at 1372
  Illegal PstI site found at 1744
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 123
  Illegal BamHI site found at 483
  Illegal BamHI site found at 1977
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 43
  Illegal PstI site found at 1372
  Illegal PstI site found at 1744
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 43
  Illegal PstI site found at 1372
  Illegal PstI site found at 1744
  
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

Figure 1- GLP-1 secretion was measured after treatment of NCI-H716 cells with anti-ICAM-2 antibodies (concentration of 1:10) for 1 h and then with P9 (50 µg ml–1) for 1 h.

In practical terms, the P9 protein can be heterologously expressed in other organisms, such as Lactococcus lactis, to produce and secrete the protein. This approach has been successfully demonstrated, where the engineered L. lactis not only secreted P9 but also stimulated GLP-1 production in NCI-H716 cells, a model for intestinal L cells. The relative expression levels of GLP-1 biosynthesis genes GCG and PCSK1 were upregulated, indicating the potential of this engineered strain for therapeutic use.

Figure 2- GLP-1 secretion by NCI-H716 cells stimulated by incubating with cell-free supernatant of L. lactis for 2 h. GM17: the supernatant of NCI-H716 cells treated by GM17 medial. NZCK: the supernatant of NCI-H716 cells treated by L. lactis NZCK. NZP9: the supernatant of NCI-H716 cells treated by L. lactis NZP9.

For users interested in incorporating the P9 protein gene into their projects, it can be used as a genetic component to develop probiotics or other biotherapeutics aimed at improving glucose metabolism and treating metabolic diseases. The gene can be codon-optimized for the host organism and expressed under controlled conditions to ensure optimal protein production and secretion.

References: Yoon HS, Cho CH, Yun MS, et al. Akkermansia muciniphila secretes a glucagon-like peptide-1-inducing protein that improves glucose homeostasis and ameliorates metabolic disease in mice. Nat Microbiol. 2021;6(5):563-573. doi:10.1038/s41564-021-00880-5

Di W, Zhang Y, Zhang X, et al. Heterologous expression of P9 from Akkermansia muciniphila increases the GLP-1 secretion of intestinal L cells. World J Microbiol Biotechnol. 2024;40(7):199. Published 2024 May 10. doi:10.1007/s11274-024-04012-z