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A natural mutation in Pisum sativum L. (pea) alters starch assembly and improves glucose homeostasis in humans

Nature Food volume 1, pages 693–704 (2020)Cite this article

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Abstract

Elevated postprandial glucose (PPG) is a significant risk factor for non-communicable diseases globally. Currently, there is a limited understanding of how starch structures within a carbohydrate-rich food matrix interact with the gut luminal environment to control PPG. Here, we use pea seeds (Pisum sativum) and pea flour, derived from two near-identical pea genotypes (BC1/19RR and BC1/19rr) differing primarily in the type of starch accumulated, to explore the contribution of starch structure, food matrix and intestinal environment to PPG. Using stable isotope 13C-labelled pea seeds, coupled with synchronous gastric, duodenal and plasma sampling in vivo, we demonstrate that maintenance of cell structure and changes in starch morphology are closely related to lower glucose availability in the small intestine, resulting in acutely lower PPG and promotion of changes in the gut bacterial composition associated with long-term metabolic health improvements.

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Fig. 1: Starch biosynthetic pathway in pea seeds.
Fig. 2: Effects of acute consumption of 50 g dry weight RR and rr pea seeds and flour.
Fig. 3: Impact of genotype structure and processing on starch digestibility.
Fig. 4: The effect of structure and genotype of pea seeds and flour on small intestinal environment.
Fig. 5: Using stable isotope 13C-enriched RR and rr pea seeds and flour to understand the digestion and fermentation process further.
Fig. 6: The effect of consuming products derived from the two pea genotypes for 28 days on glucose homeostasis and gut microbiota.

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Data availability

All presented data are tabulated and detailed in the main text and the Supplementary Information. The experimental procedures are detailed in the Methods. Quantified data are freely available from the Mendeley Data Database at https://doi.org/10.17632/gtthhhp9wz.1.

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Acknowledgements

We thank E. Panteliou and L. Mendoza for their help during nasogastric and nasoduodenal tube insertion. We thank A. Sukkar and A. Cherta Murillo for their assistance in trial 4 and G. Franco Becker and C. Byrne for their help with the radioimmunoassays in trial 4. We thank E. McKay for technical assistance with the plasma [13C]glucose and urine 13C-labelled SCFA assay and S. Small for technical assistance with the breath 13C and urine 13C-labelled SCFA assay and growth of isotope-labelled pea seeds. We thank M. Parker for valuable discussions and interpretation of the microscopy images. We thank B. Hazard, QIB, for very helpful discussions of starch mutations in cereals. All clinical trials were conducted at the NIHR Imperial Clinical Research Facility; we thank all of the staff and volunteers who took part in the study. The Division of Integrative Systems Medicine and Digestive Disease at Imperial College London receives financial support from the NIHR Imperial Biomedical Research Centre based at Imperial College Healthcare NHS Trust and Imperial College London, in line with the Gut Health research theme. I.G.-P. is supported by a NIHR fellowship (NIHR-CDF-2017-10-032). C.D. gratefully acknowledges support from the Department for Environment, Food & Rural Affairs (CH0103 and CH0111, Pulse Crop Genetic Improvement Network; and LK09126) and from the Biotechnology and Biological Sciences Research Council (BBSRC; BB/L025531/1 and BBS/E/J/000PR9799). We also gratefully acknowledge the support of the BBSRC through the BBSRC Institute Strategic Programme Food Innovation and Health BB/R012512/1 and its constituent project(s) BBS/E/F/000PR10343 (Theme 1, Food Innovation) and BBS/E/F/000PR10345 (Theme 2, Digestion in the Upper GI Tract). Infrastructure support was provided by the NIHR Imperial Biochemical Research Centre and the NIHR Imperial Clinical Research Facility. G.S.F. is an NIHR Senior Investigator. This research was funded by the BBSRC (grant nos. BB/L025582/1, BB/L025418/1, BB/L025531/1 and BB/L025566/1). The views expressed in this publication are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care.

Author information

Author notes

  1. These authors contributed equally: Katerina Petropoulou, Louise J. Salt, Cathrina H. Edwards, Frederick J. Warren, Douglas J. Morrison, Tom Preston, Claire Domoney, Peter J. Wilde, Gary S. Frost.

Authors and Affiliations

  1. Section for Nutrition Research, Department of Metabolism, Digestion and Reproduction, Faculty of Medicine, Imperial College London, London, UK

    Katerina Petropoulou, Edward S. Chambers, Rasha Alshaalan, Mai Khatib, Elaine Holmes & Gary S. Frost

  2. Quadram Institute Bioscience, Norwich, UK

    Louise J. Salt, Cathrina H. Edwards, Frederick J. Warren, Natalia Perez-Moral, Kathryn L. Cross, Lee Kellingray, Rachael Stanley, Todor Koev, Arjan Narbad & Peter J. Wilde

  3. Computational and Systems Medicine, Division of Integrated Systems Medicine and Digestive Diseases, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, London, UK

    Isabel Garcia-Perez, Nicholas Penney, Jose Ivan Serrano-Contreras & Elaine Holmes

  4. Clinical Nutrition Program, Department of Health, College of Health and Rehabilitation Sciences, Princess Noura Bint Abdulrahman University, Riyadh, Saudi Arabia

    Rasha Alshaalan

  5. Faculty of Applied Medical Sciences, Department of Clinical Nutrition, King Abdulaziz University, Jeddah, Saudi Arabia

    Mai Khatib

  6. School of Pharmacy, University of East Anglia, Norwich, UK

    Todor Koev & Yaroslav Z. Khimyak

  7. Department of Mechanical Engineering, Imperial College London, London, UK

    Maria N. Charalambides

  8. Division of Integrative Systems Medicine and Digestive Disease, Department of Metabolism, Digestion and Reproduction, Imperial College London, London, UK

    Jesus Miguens Blanco, Rocio Castro Seoane & Julian R. Marchesi

  9. MRC Centre for Molecular Bacteriology and Infection, Imperial College London, London, UK

    Julie A. K. McDonald

  10. School of Biosciences, Cardiff University, Cardiff, UK

    Julian R. Marchesi

  11. Centre for Computational & Systems Medicine, Murdoch University, Perth, Western Australia, Australia

    Elaine Holmes

  12. Diabetes, Endocrinology and Metabolism, Department of Medicine, Imperial College London, London, UK

    Ian F. Godsland

  13. Scottish Universities Environmental Research Centre, University of Glasgow, East Kilbride, UK

    Douglas J. Morrison & Tom Preston

  14. John Innes Centre, Norwich, UK

    Claire Domoney

Authors
  1. Katerina Petropoulou
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Contributions

K.P. and L.J.S. should be considered as joint first authors. G.S.F. oversaw the design and implementation of the in vivo experiments. K.P. managed, assisted in the design and performed all the experimental studies in vivo, sample collection, processing and data analysis. E.S.C. designed and applied for ethics approval for the human studies. R.A. and M.K. assisted in experimental human studies 2 and 3, and M.K. also assisted in study 4. N.P. was responsible for the nasogastric and nasoduodenal tube insertion. Metabolomics analysis was performed by K.P. and I.G.-P. Metabolite identification was performed by I.G.-P. and J.I.S.-C. D.J.M. and T.P. oversaw the stable isotope analysis and data analysis, and T.P. also oversaw the labelled crop production. L.J.S. performed simulated digestions, starch analysis of pea seeds, particle size analysis of pea fragments, sample preparation for light microscopy and scanning electron microscopy, and light microscopy imaging. Preparation, sectioning and imaging of pea tissue sections were performed by R.S. and K.L.C.; K.L.C. performed scanning electron microscopy. N.P. carried out simulated digestions of flours and subsequent starch analysis; diffusion experiments using fluorescence microscopy were also performed by N.P. R.A. performed the compression experiments and M.N.C. oversaw the implementation of these experiments. T.K. and Y.Z.K. carried out solid-state NMR experiments. P.J.W., F.J.W. and C.H.E. oversaw the design and implementation of the digestions in vitro and microscopy studies. C.D. oversaw field trials of the variant pea lines and multiplication of their seeds, with quality testing for all experiments. K.P., J.A.K.M., R.C.S. and J.M.B. performed 16S rRNA gene sequencing and data analysis. G.S.F. and K.P. led the initial drafts of the manuscript. All authors contributed to the final draft of the manuscript.

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Correspondence to Gary S. Frost.

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Supplementary Figs. 1–9, Tables 1–14, methods and note.

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Petropoulou, K., Salt, L.J., Edwards, C.H. et al. A natural mutation in Pisum sativum L. (pea) alters starch assembly and improves glucose homeostasis in humans. Nat Food 1, 693–704 (2020). https://doi.org/10.1038/s43016-020-00159-8

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