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Heterologous expression of P9 from Akkermansia muciniphila increases the GLP-1 secretion of intestinal L cells

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Abstract

Glucagon-like peptide-1(GLP-1) is an incretin hormone secreted primarily from the intestinal L-cells in response to meals. GLP-1 is a key regulator of energy metabolism and food intake. It has been proven that P9 protein from A. muciniphila could increase GLP-1 release and improve glucose homeostasis in HFD-induced mice. To obtain an engineered Lactococcus lactis which produced P9 protein, mature polypeptide chain of P9 was codon-optimized, fused with N-terminal signal peptide Usp45, and expressed in L. lactis NZ9000. Heterologous secretion of P9 by recombinant L. lactis NZP9 were successfully detected by SDS-PAGE and western blotting. Notably, the supernatant of L. lactis NZP9 stimulated GLP-1 production of NCI-H716 cells. The relative expression level of GLP-1 biosynthesis gene GCG and PCSK1 were upregulated by 1.63 and 1.53 folds, respectively. To our knowledge, this is the first report on the secretory expression of carboxyl-terminal processing protease P9 from A. muciniphila in L. lactis. Our results suggest that genetically engineered L. lactis which expressed P9 may have therapeutic potential for the treatment of diabetes, obesity and other metabolic disorders.

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Data availability

No datasets were generated or analysed during the current study.

Abbreviations

GLP-1:

Glucagon-like peptide-1

PCSK1 :

Proprotein convertase subtilisin/kexin type 1 gene

GCG :

Proglucagon gene

SCFAs:

Short-chain fatty acids

GPCR:

G-protein-coupled receptors

cAMP:

Cyclic adenosine monophosphate

PKA:

Protein kinase A

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Acknowledgements

This work was supported by the National Key R&D Program of China (No. 2022YFF1100102) and the National Natural Science Foundation of China (No. 31972055).

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Authors and Affiliations

  1. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China

    Wenxuan Di, Yuchen Zhang, Luxuan Han, Liang Zhao & Zhengyuan Zhai

  2. Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, Beijing, 100093, China

    Yanling Hao

  3. Food Laboratory of Zhongyuan, Luohe, Henan, 462300, China

    Liang Zhao, Yanling Hao & Zhengyuan Zhai

  4. Department of Food Science, Beijing University of Agriculture, Beijing, China

    Xinyuan Zhang

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  1. Wenxuan Di
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  2. Yuchen Zhang
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  4. Luxuan Han
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  5. Liang Zhao
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  6. Yanling Hao
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Contributions

W.D. (First Author): Investigation, Formal Analysis, Writing - Original Draft Y.Z.: Data Curation, Writing - Original Draft X.Z.: Visualization, Investigation L.H.: Formal Analysis L.Z.: Review & Editing Y.H.: Review & Editing Z.Z.(Corresponding Author): Conceptualization, Funding Acquisition, Resources, Supervision, Review & Editing.

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Correspondence to Zhengyuan Zhai.

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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 40, 199 (2024). https://doi.org/10.1007/s11274-024-04012-z

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