PLANT-BASED PROTEIN MIXTURES AND NUTRITIONAL COMPOSITIONS

Abstract

The present invention relates to plant-based protein mixture and nutritional compositions and supplements comprising said mixtures.

Claims

1. A plant-based protein mixture comprising: a) 50.0%-95.0% w/w pulse protein b) 5.0%-50.0% w/w or more non-pea plant protein based on total protein content and 60.0%-25.0% w/w essential amino acids based on total protein content.

2. The mixture according to claim 1, wherein said mixture comprises 50.0%-25.0% w/w essential amino acids based on total protein content.

3. The mixture according to claim 1, wherein said mixture comprises at least 5.0%-15.0% w/w, leucine based on total protein content.

4. The mixture according to claim 1 having a PDCAAS of 0.95 or higher.

5. The mixture according to claim 1 that is substantially free of cereal, gluten, soy and/or dairy matter.

6. The mixture according to claim 1, wherein the pulse protein is pea protein.

7. The mixture according to claim 1, wherein said proteins are substantially intact proteins.

8. A mixture according to claim 1 in a powdered, spray dried or granule form.

9. A method for protein supplementation of an individual comprising administering to the individual a composition according to claim 1.

10. The method according to claim 9, wherein said composition is ingested prior to during and/or subsequent to physical exercise.

11. A method for improving a result of physical exercise in a subject, comprising administering to the subject a composition according to claim 1.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0077] FIG. 1a. Shows mean of blood eAA over 4 hours per time point. FIG. 1b. Shows mean and 95% CI total sum blood eAA iAUC (nmol/mL) over 4-hours as described in Example 1. The area under the curve above baseline vs. time (min) was obtained by using the trapezoidal rule.

[0078] FIG. 2a. Shows mean of blood eAA over 4 hours per time point. FIG. 2b. Shows mean and 95% CI total sum blood eAA iAUC (nmol/mL) over 4-hours as described in Example 1. The area under the curve above baseline vs. time (min) was obtained by using the trapezoidal rule.

[0079] FIG. 3 provides an overview of the protein supplement preparation process as described in Example 1.

EXAMPLES

[0080] Objectives

[0081] The inventors hypothesized that the complete plant-based protein blends of the invention, with a 1.0 PDCAAS, would be bioequivalent (defined in this study as similar blood eAA response) to WPI.

Methods

[0082] The primary objective of the study (ClinicalTrial.gov no. NCT03673839) was to assess the bio-equivalence of the total blood eAA response (iAUC) over 4 hours. The secondary objectives were to assess the bio-equivalence (iAUC) and leucine kinetics (Cmax and Tmax) over 4 hours.

[0083] Participants: The study was an acute, randomized, double-blind, cross-over, 4 periods×4 products William's square design study. Participants were healthy, adult men, 18-35 years of age, Body Mass Index (BMI) between 18.5 and 29.9 kg/m2. Participants were required to have self-reported resistance training experience of no less than 2 years, with resistance training of at least 1 hr/day for 2 days/week over the past 6 months.

[0084] Visits: Participants were instructed to abstain from any protein supplements for 1 day prior to the study intervention phase. Prior to each study visit, the participants were required to consume a standardized dinner, followed by at least a 12-hour overnight fast. Participants were randomized by sequence, on each study visit the participants consumed one of four study products: three plant-based protein blends (Tests) and a WPI (Control), mixed with water. One of the three plant-based blends was a hydrolysate treated with a protease enzyme.

[0085] The participants were given a study beverage and instructed to consume it over 10 minutes. Blood samples were collected before product consumption and 15 min, 30 min, 1 h, 2 h, 3 h and 4 h after the consumption of the study beverage. Blood amino acids were measured by ultraperformance liquid chromatography (UPLC).

[0086] Statistics: After a log-transformation of the total sum of blood eAA's iAUC over 4 hours, a repeated measures ANOVA model with fixed effect terms for sequence, product, period and participant within sequence as random was fitted to assess for bio-equivalence. Back transformed LS-Means differences and 90% CIs (i.e., ratio of Test to Control Geometric (GLS-Means) and associated 90% CI). For bio-equivalence to be demonstrated, the entirety of the 90% CI for the ratio of Test to Control GLS-Means must lie within the region (0.80 to 1.25).

Test Product

[0087] There were 4 study protein supplement test products, all of which were powdered drink mixes provided in single serving sachets to be mixed into 12 oz. of water. The study products consisted of animal (control) and plant-based proteins (3 test products) in a sweetened flavor system (cocoa powder, flavoring agents, stevia, xanthan gum). The control product was a commercially available Whey Protein Isolate. For the study, the product was purchased and repackaged into study product packaging.

[0088] All three plant-based blends included: [0089] (i) Pea protein “Organic Puris pea 870H” (PurisPea, Minneapolis, Minn.; protein content: 80% weight); and [0090] (ii) Pumpkin protein “Organic unroasted Pumpkin protein” (Austrade Inc., Palm Beach Gardens, Fla.; protein content: 60% weight).

[0091] Blend #3 was exactly the same as blend #1. However, blend #3 was a hydrolysate.

[0092] Blend #2 contained, in addition to the pea and pumpkin protein:

[0093] Sunflower protein “Helifor 55” (Austrade Inc., Palm Beach Gardens, Fla.; protein content: 55% weight); and

[0094] Coconut protein “Organic Coconut Protein 17” (Austrade Inc., Palm Beach Gardens, Fla.; protein content: 17% weight).

[0095] Further blend details are provided in Table 2, and an overview of the protein supplement preparation process is provided in FIG. 3.

[0096] All study products were served as a study beverage; 33-34 g of study product (protein mixture) was mixed with 12 oz (360 g) of bottle water of same brand in a shaker bottle. All study products were similar in appearance, packaging and nutritional content to maintain the double-blind nature of the study.

[0097] Study products were consumed within 10 minutes by participants.

[0098] In addition, after consumption of the study beverage, subjects were also given 50 ml of water to wash out the bottle and clean their mouth, ensuring that all product was consumed.

[0099] The storage of study products was at room temperature.

TABLE-US-00002 TABLE 2 Blend 2 Pea 82% Blend 3 Whey Blend 1 Pumpkin 4% Hydrolysate Protein (+ Pea 76% Sunflower 11% Pea 76% control) Pumpkin 24% Coconut 3% Pumpkin 24% Total Protein 24 g 34 g 33 g 34 g Leucine 2.6 g 2.6 g 2.6 g 2.6 g Total 5.6 g 5.6 g 5.8 g 5.6 g BCAAs Total eAAs 13.2 g 13.8 g 13.9 g 13.8 g PDCAAS 1.0 1.0 1.0 1.0

Blends 1&3

[0100] Pea “Organic Puris pea 870H” 76 g per 100 g protein powder mixture—60.8 g protein (76*0.8)

[0101] Pumpkin “Organic unroasted Pumpkin protein” 24 g per 100 g protein powder mixture—14.4 g protein (24*0.6).

[0102] Thus Blends 1 & 3 contained 81% w/w pulse protein relative to total protein content and 19% w/w non-pea plant protein relative to total protein content.

Blend 2

[0103] Pea “Organic Puris pea 870H” 82 g per 100 g protein powder mixture—65.6 g protein (82*0.8)

[0104] Pumpkin “Organic unroasted Pumpkin protein” 4 g per 100 g protein powder mixture—2.4 g protein (4*0.6).

[0105] Sunflower “Helifor 55” 11 g per 100 g protein powder mixture—5.5 g protein (11*0.5)

[0106] Coconut “Organic Coconut Protein 17” 3 g per 100 g protein powder mixture—0.51 g protein (3*0.17)

[0107] Thus Blend 2 contained 89% w/w pulse protein relative to total protein content and 13% w/w non-pea plant protein relative to total protein content.

Results

[0108] Eighteen men (Per Protocol set) aged 25.4±4.64 years (mean±sd) with a BMI of 24.4±3.35 kg/m.sup.2 completed the study. For the primary outcome, the total sum of eAA iAUC over 4 hours of plant-based protein blends were not bio-equivalent to WPI, the three ratios [90% CI]: Blend #1: 0.66 [0.58-0.76]; Blend #2: 0.71 [0.62-0.82], Blend #3: 0.60 [0.52-0.69], indicative of a .sup.˜29 to 40% decrease of plant-based protein blends eAA iAUC over 4 hours compared to WPI. For the secondary outcomes, the sum total of leucine iAUC over 4 hours of plant-based protein blends were not bio-equivalent to WPI, the three ratios [90% CI]: Blend #1: 0.66 [0.59-0.73]; Blend #2: 0.67 [0.61-0.75]; Blend #3: 0.62 [0.56-0.69].

[0109] Leucine Cmax of plant-based protein blends were not bio-equivalent to WPI Blend #1: 0.70 [0.67-0.73]; Blend #2: 0.72 [0.68-0.75], Blend #3: 0.65 [0.62-0.68], indicative of a .sup.˜28 to 35% decrease over 4 hours compared to WPI. Leucine Tmax for Blend #1 and Blend #3 were close to WPI (Blend #1: 0.94 [0.73-1.18]; Blend #2: 1.56 [1.28-1.92]; Blend #3: 1.19 [0.95-1.48]).

TABLE-US-00003 TABLE 3 Leucine Tnnax over 4 hours Leucine Cnnax over 4 hours (min) (SD) (nnnnol/nnl) (SD) Blend 1 43.4 (16.5) 460 (47)* Blend 2 67.9 (29.7)* 507 (63)* Blend 3 53.7 (27.5) 466 (48)* Control (WPI) 44.2 (15.4) 716 (103) *p-value<.001, pairwise Student t test of the LS-Means difference Tests compared to control.

[0110] No adverse events related to the study product intake was observed.

CONCLUSIONS

[0111] This study represents the first human investigation in which blood eAA responses to complete, plant-based protein blends (PDCAAS=1.0), matched for leucine content were compared to whey protein. The results show that the three plant-based protein blends were not bio-equivalent to the WPI control, as measured by blood eAA data. However, the leucine kinetic data across the plant-based protein blends showed an approximate 2-fold increase from fasting levels.

[0112] Further, similar leucine Tmax data were observed across two plant-based protein blends (#1 and #3) compared to WPI. From a physiological standpoint, the leucine data provide evidence of a rapid hyperleucinemia and this is a critical response associated with postprandial stimulation MPS.