The composition and nutritional quality of biorefined lucerne protein depend on precipitation method.

BACKGROUND: Lucerne protein extract is a novel high-quality protein source with excellent amino acid (AA) composition of interest for human consumption. In this study, protein from screw-pressed lucerne juice was extracted by different precipitation methods to evaluate the effect on the chemical composition and nutritional quality of the extracted protein. Methods based on heat, acidification or fermentation were used for protein precipitation, and the nutritional value of protein was evaluated in a rat digestibility trial. RESULTS: Heat precipitation at 85 °C produced a protein product with a crude protein (CP) content of 589 g kg-1 dry matter (DM), a balanced AA composition and a high standardized nitrogen (N) digestibility (82.8%). Precipitation by acidification, at a lower temperature (60 °C) or by fermentation, resulted in lower CP content (425-488 g kg-1 DM). Nitrogen digestibility for the pH-adjusted precipitate was equal to the 85 °C heat-precipitated protein, while the fermented and 60 °C precipitated proteins showed lower N digestibility (76.5% and 78.6%, respectively). By applying a two-step heat precipitation method (60 °C followed by 80 °C), a protein content of 712 g kg-1 DM and an N digestibility of 93.6% was reached, which are comparable to high-quality animal-based protein sources such as milk, whey, casein, and eggs, covering the AA requirements for children >6 months. CONCLUSION: High-quality protein can be extracted from lucerne, but the future focus should be on increased yield as the current low yields of the refined product will challenge the environmental and economic sustainability of production. © 2023 Society of Chemical Industry.

Effects of dairy and plant protein on growth and growth biomarkers in a piglet model.

The increasing world population with improved living conditions has increased the demand for food protein. This has intensified the search for sustainable alternative plant-derived high-quality protein sources for human nutrition. To study the effect of plant and milk proteins on growth in weaned pigs as a model for humans, 96 weaned pigs were divided into 48 pens and fed one of 4 different diets for 3 weeks. The dietary protein originated from either 50% rice + 50% 00-rapeseed protein (RICE + RAPE), 50% milk protein (MPC) + 50% 00-rapeseed protein (MPC + RAPE), 50% milk + 50% rice protein (MPC + RICE), or 100% MPC, and were supplemented with crystalline amino acids to meet the amino acid requirements. Weekly feed intake and body weights were recorded and after 3 weeks, a blood sample was taken 1 hour after a fixed meal, while organ weights were measured, and liver- and muscle tissue, and bone samples were collected at euthanasia. All pigs had a high daily gain and a low feed-to-gain ratio (F : G, feed intake per kg weight gain), but feed intake and daily gain was lowest and F : G highest in the RICE + RAPE diet. Metacarpal bones were longer and heavier in MPC + RICE and MPC fed pigs compared to pigs fed diet RICE + RAPE (P < 0.05), and intermediate in MPC + RAPE fed pigs, with no differences in bone thickness (P > 0.05). Plasma levels of all essential amino acids except Cys and Lys decreased markedly when fed a diet containing only plant protein. The differences were not associated with differences in plasma insulin or IGF-1, nor in the abundance of mRNA related to growth in liver and longissimus dorsi muscle. In conclusion, the growth of piglets fed a combination of milk and rice protein did not differ from the pure dairy-based diet, whereas the pure plant-based diet consisting of rice and rapeseed protein led to reduced growth. This was most likely caused by a lower feed intake and a lower than expected amino acid digestibility of the 00-rapeseed protein. There were no indications that the milk protein, beyond a favourable amino acid composition and high digestibility, specifically stimulated growth factors or other biomarkers of growth via the IGF-1 and insulin signalling pathways.

Impact of Diet-Modulated Butyrate Production on Intestinal Barrier Function and Inflammation.

A major challenge in affluent societies is the increase in disorders related to gut and metabolic health. Chronic over nutrition by unhealthy foods high in energy, fat, and sugar, and low in dietary fibre is a key environmental factor responsible for this development, which may cause local and systemic inflammation. A low intake of dietary fibre is a limiting factor for maintaining a viable and diverse microbiota and production of short-chain fatty acids in the gut. A suppressed production of butyrate is crucial, as this short-chain fatty acid (SCFA) can play a key role not only in colonic health and function but also at the systemic level. At both sites, the mode of action is through mediation of signalling pathways involving nuclear NF-κB and inhibition of histone deacetylase. The intake and composition of dietary fibre modulate production of butyrate in the large intestine. While butyrate production is easily adjustable it is more variable how it influences gut barrier function and inflammatory markers in the gut and periphery. The effect of butyrate seems generally to be more consistent and positive on inflammatory markers related to the gut than on inflammatory markers in the peripheral tissue. This discrepancy may be explained by differences in butyrate concentrations in the gut compared with the much lower concentration at more remote sites.

Postprandial PYY increase by resistant starch supplementation is independent of net portal appearance of short-chain fatty acids in pigs.

Increased dietary fiber (DF) fermentation and short-chain fatty acid (SCFA) production may stimulate peptide tyrosine-tyrosine (PYY) secretion. In this study, the effects of hindgut SCFA production on postprandial PYY plasma levels were assessed using different experimental diets in a porto-arterial catheterized pig model. The pigs were fed experimental diets varying in source and levels of DF for one week in 3×3 Latin square designs. The DF sources were whole-wheat grain, wheat aleurone, rye aleurone-rich flour, rye flakes, and resistant starch. Postprandial blood samples were collected from the catheters and analyzed for PYY levels and net portal appearance (NPA) of PYY was correlated to NPA of SCFA. No significant effects of diets on NPA of PYY were observed (P > 0.05), however, resistant starch supplementation increased postprandial NPA of PYY levels by 37 to 54% compared with rye-based and Western-style control diets (P = 0.19). This increase was caused by higher mesenteric artery and portal vein PYY plasma levels (P < 0.001) and was independent of SCFA absorption (P > 0.05). The PYY levels were higher in response to the second daily meal compared with the first daily meal (P < 0.001), but similar among diets (P > 0.10). In conclusion, the increased postprandial PYY responses in pigs fed with different levels and sources of DF are not caused by an increased SCFA absorption and suggest that other mechanisms such as neural reflexes and possibly an increased flow of digesta in the small intestine may be involved. The content of DF and SCFA production did not affect PYY levels.

Whole Grain Consumption Increases Gastrointestinal Content of Sulfate-Conjugated Oxylipins in Pigs - A Multicompartmental Metabolomics Study.

The effects of increased intake of dietary fiber as either whole grain rye or as resistant starch compared to a typical low dietary fiber Western-style diet on the metabolomics responses were studied in gastrointestinal content and tissue, peripheral plasma, and urine using a multicompartmental nontargeted liquid chromatography-mass spectrometry (LC-MS) approach in pigs. Both unsupervised and supervised multivariate analyses were used to study differences between the intervention groups, revealing significant effects of the dietary intervention on gastrointestinal contents and urine metabolites. Consumption of a diet composed of whole grain rye flakes and enzyme treated wheat bran increased gastrointestinal content of novel sulfate- and acetic acid-conjugated oxylipins, and urinary secretion of phenolic compounds. Furthermore, potential gastrointestinal and urinary biomarkers for consumption of resistant starch, whole grain rye, and a low dietary fiber intake were detected using multiblock analysis. This study provides insight into microbial fermentation products in the gastrointestinal tract and suggests a potential role in sulfate conjugation of metabolites on the bioavailability of ingested nutrients.

Resistant starch and arabinoxylan augment SCFA absorption, but affect postprandial glucose and insulin responses differently.

The effects of increased colonic fermentation of dietary fibres (DF) on the net portal flux (NPF) of carbohydrate-derived metabolites (glucose, SCFA and, especially, butyrate), hormones (insulin, C-peptide, glucagon-like peptide 1 and glucose-dependent insulinotropic peptide) and NEFA were studied in a healthy catheterised pig model. A total of six pigs weighing 59 (SEM 1·6) kg were fitted with catheters in the mesenteric artery and in the portal and hepatic veins, and a flow probe around the portal vein, and included in a double 3 × 3 cross-over design with three daily feedings (at 09.00, 14.00 and 19.00 hours). Fasting and 5 h postprandial blood samples were collected after 7 d adaptation to each diet. The pigs were fed a low-DF Western-style control diet (WSD) and two high-DF diets (an arabinoxylan-enriched diet (AXD) and a resistant starch-enriched diet (RSD)). The NPF of insulin was lower (P= 0·04) in AXD-fed pigs (4·6 nmol/h) than in RSD-fed pigs (10·5 nmol/h), despite the lowest NPF of glucose being observed in RSD-fed pigs (203 mmol/h, P= 0·02). The NPF of total SCFA, acetate, propionate and butyrate were high, intermediate and low (P< 0·01) in AXD-, RSD- and WSD-fed pigs, respectively, with the largest relative increase being observed for butyrate in response to arabinoxylan supplementation. In conclusion, the RSD and AXD had different effects on the NPF of insulin and glucose, suggesting different impacts of arabinoxylan and resistant starch on human health.