Impact of food processing on the blood cholesterol-lowering effect of cereal beta-glucan

Lead Research Organisation: University of Nottingham
Department Name: Sch of Biosciences

Abstract

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Technical Summary

The effect of beta-glucan (BG), a water-soluble form of fibre, in lowering LDL cholesterol concentrations has been demonstrated by many research groups. This beneficial impact has been accepted as a valid health claim for BG, in non-processed or minimally processed forms of oats and barley, by EFSA. The mechanism(s) of action of BG are strongly linked to its capacity to generate high viscosity in the gastrointestinal tract and a subsequent decrease in bile reabsorption through decreases in rates of digestion, inhibition of lipolysis and/or transport of mixed micelles. The BG may also directly interact with lipase, bile salts and the mucus layer, leading to decreased bile recycling. However, the precise mechanisms are still not fully understood. Moreover, the cholesterol-lowering activity of BG varies significantly depending on food type, processing and storage conditions, mainly due to depolymerisation and poor solubility of BG.

The main aims of this project therefore are to understand (a) the mechanism(s) by which BG lowers blood cholesterol and (b) how the processing of BG-containing foods attenuates this capacity of the polymer. A multidisciplinary approach involving a range of in vitro methods, biochemical (e.g. in vitro digestion, enzyme kinetics) and biophysical (e.g. rheology, fluorescence microscopy), will be used. Samples of purified BG and BG-containing cereals and real foods (porridge & biscuits) will be characterised (e.g. molecular weight, polymer dissolution), before and after food processing, and their behaviour during in vitro digestion and their interactions with BS and lipase(s), will be studied. These samples will be subjected to a range of sophisticated in vitro digestion models allowing viscosity and interactions to be determined during breakdown and transport. This work will provide new information on how BG can be processed and added to food matrices in a way that maintains optimal health effects.

Planned Impact

The proposed project will benefit relevant stakeholders outside the academic communities, including the commercial sector, public sector bodies and the general public. It addresses one of the UK's major diet-related health challenges, cardiovascular disease (CVD). The project objective is also relevant to the BBSRC strategy priorities as it will "improve our understanding of the links between diet and health" by "optimisation of product formulation through new uses of existing ingredients, novel ingredients and novel formulations of ingredients." It therefore has huge potential long-term impact on a wide range of stakeholders from consumers to policy makers.

The number of individuals diagnosed with CVD in the UK has remained roughly static over the last 15-20 years at 5% of the adult population with 35% of cases being over the age of 55. Raised blood cholesterol is an important risk factor for CVD. Each year more than 150,000 deaths are caused by CVD in the UK. Associated with this alarming death rate, the annual financial cost exceeds £30bn, with cholesterol-lowering drugs alone costing the NHS approximately £500 million. Dietary interventions appear to be more appealing solutions, especially since this approach may simultaneously have positive effects on other diet-related conditions such as obesity and type II diabetes.

Our work will be particularly useful to the food industry and may lead to innovative applications in the future, such as the development of 'functional ingredients and foods' that have the capacity to consistently reduce blood cholesterol concentrations. The design of foods made from oats or barley beta-glucan would benefit public health. The main beneficiaries will be the DRINC members, especially those that are involved in the processing of oats and barley.

Individual consumers will have new knowledge regarding the health benefits of certain fibre-containing foods, together with a wider choice of manufactured foods with specific, proven health benefits. However, care has to be taken with the health communication aspects, since the consumption of these foods may only be effective as part of a healthy balanced diet. The foods ought to have comparable consumer acceptability, making it more convenient for consumers to adopt healthier dietary options. The long-term health benefits of reducing obesity and the risks associated with diabetes will benefit the lifelong health and well-being of individuals and improve their quality of life into old age.
The research should also help policy-makers in formulating new dietary advice and guidelines from new data on plant foods (oats and barley). The applicants regularly give talks at schools and media interviews about their research.

The applicants at the three academic centres will have regular meetings to discuss research findings. The consortium of food companies in DRINC will be offered first-market access to intellectual property if the results of the research lead to industrial applications. All the academic centres at KCL, IFR and Nottingham have mechanisms in place for exploring industrial applications from the research work. The applicants, research staff and PhD student will all play some role in undertaking impact activities. Many of the applicants have relevant experience for achieving successful knowledge exchange and impact with the beneficiaries. The applicants have also a long history of Knowledge Exchange and Commercialisation activities, particularly with the food industry, including industrial grants, patents, exploitation activities and dissemination events.

Publications

10 25 50

 
Description The contribution from the NCMH (National Centre for Macromolecular Hydrodynamics) team at Nottingham (BBSRC DRINC grant BB/L025477/1), award £66,469, is part of a 3-way collaboration/ project - with the Quadram Institute of Bioscience at Norwich (PI Prof. Peter Wilde) - BB/L025299/1 - and the Biopolymers Group at King's College London BB/L025272/1 with Professor Peter Ellis as PI and the Lead investigator. The £66,469 support for the NCMH included 5.5 months of a research technician (Dr. Guy Channell). The full report has been made by Professor Ellis in his Research Fish entry, but we take the opportunity to focus of the NCMH's impact. Beta-glucans from oats and other cereals are a major source of soluble dietary fibre, and which have other beneficial properties. One of these is cholesterol level lowering. No-one is quite sure how this happens, but one of the key physical properties is their molecular weight or molecular weight distribution. Because of the difficult nature of characterising these parameters we adapted a method that we designed and published in 2011 - the Extended Fujita Method - based on analytical ultracentrifugation - for achieving this. It is a particularly suited for analysing samples of non-perfect purity, and this we did: establishing the parameters with a well characterised beta glucan (BG90) and then successfully applying the method to both impure preparations containing as little as 32% beta glucan and roll-mill processed beta glucans. This work was submitted to Scientific Reports for publication and has been published (see link below). This also required a key refinement to the way samples are loaded into the analytical ultracentrifuge for analysis by establishing a simple and accurate method for aligning the "cells" which contain the samples for analysis. That work - which was presented at the Analytical Ultracentrifuge conference in Glasgow and has also been published in European Biophysics Journal (see link below). The next task was to explore the effect of thermal and freeze processing (autoclave, freeze thaw) on the molecular integrity (molecular weight) of BG90 and this we did using also dynamic and classical light scattering and viscosity. Gratifyingly the beta glucans appear largely robust based on our accurate molecular weight distribution procedures. That work will be submitted shortly for publication (see link below). The fourth paper in the series uses the potential of the analytical ultracentrifuge to probe interactions through the principle of co-sedimentation. There appears to be a significant interaction and further final experiments are being completed by a PhD student to quantify its strength and this will be submitted for publication (see below). Another paper is now in preparation (in conjunction with NOFIMA - The Norwegian Institute of Food, Fisheries and Aquaculture Research, at Ås) titled "Inter-laboratory evaluation of different approaches for the determination of weight-average molecular weight and molecular weight distribution of cereal beta-glucan extracts", also expected to be submitted shortly.
Exploitation Route Our findings have fed into the work of the other groups involved the project at Kings College London and at Quadram in Norwich. They too have produced significant papers focusing on their input: with Prof Ellis (project lead) who has pulled together the work into a final report. The lipase interaction studies through Nottingham PhD student Yundong Lu (at no extra cost to the BBSRC), who is doing great work . A summary of the work so far including the 4 papers that have directly come from the work of the Nottingham team can be found on our links page: https://www.nottingham.ac.uk/-sczsteve/Links%20to%20the%20papers.htm. The work has formed the basis of a follow up application on beta-glucan and other dietary fibre interactions with gut mucus/ mucins "Molecular defence in the mammalian gut: Unlocking the mechanisms of dietary fibre interaction with mucin glycoproteins" with Dr. G. Yakubov that was funded. The knowledge gained about beta-glucans will prove invaluable in this next BBSRC project which will help us understand how beta glucan and other dietary fibre interacts with gut mucus to give their huge beneficial effects.
Sectors Agriculture, Food and Drink,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

URL https://www.nottingham.ac.uk/-sczsteve/Links%20to%20the%20papers.htm
 
Description Our findings have fed into the work of the other groups involved the project at Kings College London and at Quadram in Norwich: please refer to the parent report by Professor Peter Ellis (project lead) written on all our behalf who has pulled together the work into a final report. Once the lipase studies by Nottingham MRes student Yundong Lu are completed (NOT funded by the BBSRC) this will give further impact as we will be able to confirm or rule out the role of lipases in the cholesterol process. This is important and will give confidence to the public who are using beta-glucans in the knowledge that we know better how these materials can reduce their cholesterol levels. The general public would we are sure be delighted to know that the work has also formed the basis of a follow up application on beta-glucan and other dietary fibre interactions with gut mucus/ mucins "Molecular defence in the mammalian gut: Unlocking the mechanisms of dietary fibre interaction with mucin glycoproteins" with Dr. G. Yakubov that was recently funded by the BBSRC: this will ultimately reveal the how the mucus lining our alimentary tracts works with beta glucans and other dietary fibre, in keeping us healthy!.
First Year Of Impact 2020
Sector Agriculture, Food and Drink
Impact Types Societal

 
Title Cell alignment procedure for the analytical ultracentrifuge facilitating accurate molecular weight & interaction analysis 
Description A key tool for analysing the molecular weight distribution of beta glucans - and for exploring their interactions with lipases is the Analytical Ultracentrifuge. The accuracy with tis this instrument can provide molecular weight and interaction parameters and information depends on the alignment of the special "cells" which contain the samples. We establish a simple and accurate method of doing this. Here is the abstract of the paper currently accepted (subject to minor revision) by European Biophysics Journal: "In establishing the sources of data variability within SV-AUC analysis and their relative importance recent studies have demonstrated that alignment of the sample cells to the centre of rotation is the most significant contributing factor to overall variability. In this study we demonstrate the usefulness of a simple manual alignment protocol with the aid of x10 magnifying eye-piece and show it is possible to approach the performance of more sophisticated mechanical alignment tools. Using a 0.7mg/ml bovine serum albumin solution a range of off-set angles were investigated and compared. The effect of increased mis-alignment angle is shown to cause an increased peak broadening for both the BSA monomer and dimer peaks whereas only the dimer shifts to lower sedimentation coefficients. Reducing the misalignment to within +1 degree of true alignment minimizes these effects and we show this is possible using both our manual alignment protocol and commercially available manual alignment procedures". 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? Yes  
Impact This procedure has been used for the characterisations in the project, and its impact relates to the accuracy which we measure molecular weights and interactions. It will also help in other projects, not just at the NCMH at Nottingham but also for other users of this instrumentation. 
URL https://www.nottingham.ac.uk/-sczsteve/Channell%20et%20al%202018c.pdf
 
Title Extended Fujita Method for characteristing Beta-Glucans 
Description Molecular weight and molecular weight distribution are the 2 most important physical properties of macromolecules - particularly beta-glucans which are the subject of this project. The Extended Fujita Method involves use of sedimentation velocity in the analytical ultracentrifuge to accurately define the molecular weight distribution of polymers without complications of fractionation columns or membranes, and with a huge range of sizes possible. Originally published in 2011 (HARDING, S.E., SCHUCK, P., ABDELHAMEED, A.S., ADAMS, G.G., KÖK, M.S. and MORRIS, G.A., 2011. Extended Fujita approach to the molecular weight distribution of polysaccharides and other polymeric systems Methods. 54(1), 136-144 http://dx.doi.org/10.1016/j.ymeth.2011.01.009 we have successfully adapted and set the parameters for beta glucans, important for the success of the project. 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? Yes  
Impact We have successfully adapted our Extended Fujita procedure so as to provide an accurate tool for characterising molecular weights and molecular weight distributions of beta-glucans in solution, with the following article now submitted to Scientific Reports for consideration: https://www.nottingham.ac.uk/-sczsteve/Channel%20et%20al%202018a.pdf 
URL https://www.nottingham.ac.uk/-sczsteve/Channell%20et%20al%202018a.pdf
 
Description Collaboration with Nofima AS, the Norwegian Institute of Food, Fisheries and Aquaculture Research, Ås (near Sarpsorg) Norway 
Organisation NOFIMA Ås
Country Norway 
Sector Private 
PI Contribution A major partnership has been made with the Nofima AS, the Norwegian Institute of Food, Fisheries and Aquaculture Research, based at Ås (near Sarpsorg) Norway. This is through Dr. Simon Ballance. We have provided our hydrodynamic expertise in characterising complex macromolecular system of beta glucans, namely analytical ultracentrifugation (sedimentation velocity and equilibrium). A scientific paper "Use of the Extended Fujita method for representing the molecular weight and molecular weight distributions of native and processed oat beta-glucans" has currently been submitted for publication to Scientific Reports with and a second one - based on standardisation of a beta glucan from which other beta glucans can be characterised against - is in preparation.
Collaborator Contribution Simon's group at Nofima have helped source materials for the project and also have provided a complementary range of characterisation expertise. In particular they have an independent set up of characterisation equipment know as "SEC MALS" (size exclusion chromatography coupled to light scattering) - and as a perusal of the URL will show we were able to get identical results, hence validating each others equipment: very important considering the difficult nature of the substance - beta glucans - at the centre of the project. Simon has also interacted with
Impact (1) Research paper: "Use of the Extended Fujita method for representing the molecular weight and molecular weight distributions of native and processed oat beta-glucans" - currently submitted to Scientific Reports. (2) A 2nd is in preparation (involving also collaboration with groups at the NTH Trondheim, Norway (Prof. B.E. Christensen) and Dr. Marius Schmidt (Coriolis-Pharma, Planegg, Germany)
Start Year 2016
 
Description Macromolecular interactions at the mucin/ mucosal interface 
Organisation University of Leeds
Department School of Food Science and Nutrition Leeds
Country United Kingdom 
Sector Academic/University 
PI Contribution The collaboration is with Professor Alan Mackie's Group which has resulted from this grant. The collaboration has involved macromolecular interactions at the mucosal interface. This may influence the way beta glucans exert their benefical effect of reducing cholesterol levels. With Mackie we have been exploring mucosal interactions on some well understood soluble dietary fibre polysaccharides. We have provided our hydrodynamic expertise in characterising both mucins and polysaccharides The collaboration started whilst Prof Mackie was at the Quadram Institute of Bioscience, and has continued since Mackie has moved to become Chair of Food Science at the University of Leeds.
Collaborator Contribution Prof Mackie's group have isolated and purified the mucins
Impact 2 papers (1) ALMUTAIRI, F.M., HERNANDEZ-CIFRE, J.G., ADAMS, G.G., KOK, M.S., MACKIE, A.R., GARCIA DE LA TORRE, J. and HARDING, S.E., 2016. Application of recent advances in hydrodynamic methods for characterising mucins in solution European Biophysics Journal. 45(1), 45-54 http://dx.doi.org/10.1007/s00249-015-1075-0 (2) MACKIE AR, MACIERZANKA A, AARAK K, RIGBY NM, PARKER R, CHANNELL GA, HARDING SE and BAJKA BH, 2016. Sodium alginate decreases the permeability of intestinal mucus. Food Hydrocolloids. 52, 749-755 http://www.sciencedirect.com/science/article/pii/S0268005X15300497
Start Year 2015