Gut microbiota modulation and amino acid absorption by Lactiplantibacillus plantarum TWK10 in pea protein ingestion: TWK10 boosts hut microbiota, amino acid uptake

Abstract

For vegetarians or vegan athletes, improving the utilization of plant-based protein and the absorption of amino acids is crucial. This study explored the impact of combining pea protein with Lactiplantibacillus plantarum TWK10 and resistance training on amino acid absorption and exercise performance. Sixteen male and sixteen female participants were randomly assigned to either a control group (20 g of pea protein without TWK10) or a TWK10 group (20 g of pea protein combined with 1 × 10¹⁰ colony-forming units of TWK10). After 28 days of supplementation combined with resistance exercise training three times per week. All subjects underwent body composition and muscle strength performance, plasma and fecal samples were collected for microbiota analysis and blood amino acid concentrations. The TWK10 group showed a significant increase in muscle thickness and improvements were observed in 1 repetition maximum bench press, explosive, anaerobic power output compared to before the intervention, and were significantly higher than those in the control group (p < 0.05). TWK10 supplementation significantly increased the area under the curve and maximum concentration of branched-chain amino acids, essential amino acids, and total amino acids (p < 0.05). Furthermore, TWK10 supplementation effectively increased the richness of gut bacterial families. Our study demonstrated that the TWK10 significant increase in the abundance of specific bacterial families in the gut, resulting in increased pea protein amino acid absorption. Moreover, increasing muscle mass and significantly improving muscle thickness, muscle strength, power, and anaerobic capacity.

Keywords: Branched-chain amino acids; Exercise performance; Muscle strength; Probiotic; Supplement.

Graphical abstract

Fig. 1

Experimental design.

Fig. 2

Effects of TWK10 administration on (A) right biceps muscle thickness and change, (B) right quadriceps muscle thickness and change, (C) left biceps muscle thickness and change, (D) left quadriceps muscle thickness and change, (E) right biceps fascia thickness and change, (F) right quadriceps fascia thickness and change, (G) left biceps fascia thickness and change, and (H) left quadriceps fascia thickness and change. Data are shown as mean ± SD. Treatment effect was analyzed by unpaired Student's t-test. ^# presented a significant effect by repeated measures ANOVA (p < 0.05). The changes in muscle and fascia thickness were calculated as the difference between after and before administration, and statistical significance was analyzed using the Mann-Whitney U test. †p < 0.05.

Fig. 3

Effects of TWK10 administration on exercise and strength performance (A) 1-RM bench press test and change (B) IMTP test of peak force and change, (C) IMTP test pRFD and change, (D) Wingate test of relative mean power and change, (E) Wingate test of relative peak power and change, and (F) Wingate test of fatigue index and change. Data are shown as mean ± SD. Treatment effects on Relative mean power, Relative peak power, and Fatigue index were analyzed by Mann-Whitney U test. ^# presented a significant effect by repeated measures ANOVA (p < 0.05). The changes in Relative mean power, Relative peak power, and Fatigue index were analyzed using the Mann-Whitney U test. ^†p < 0.05.

Fig. 4

Alpha and beta diversity of microbial communities. (A, B) Box plots illustrate the differences in alpha-diversity indices (observed ASVs and Shannon index) between the control and TWK10 groups before and after administration, (C) NMDS ordinations based on Bray-Curtis distances between all samples at the ASV level. Differences on the NMDS plot between the values of each group were analyzed by PERMANOVA, and (D) Box plots show within treatment group variation of beta diversity measures for Bray-Curtis dissimilarity. Comparison of bacterial abundance between before and after administration in the control and TWK10 groups. The bacterial composition is based on the (E) top 10 phyla and (F) top 25 families of bacteria in all samples. Others represent the remaining phyla or families with lower relative abundance. Differential abundance analyses at the family level between before and after administration in the control (G) and TWK10 (H) groups by ANCOM-BC. Dynamic signature of gut microbiota components in the TWK10 group. (I) Differential abundance analyses at the genus level between before and after administration in the TWK10 group by ANCOM-BC. (J) Arrows show genera significantly correlated with the NMDS axes, which were determined by the envfit function of the R package vegan, with a cut-off for plotted results of p < 0.01 and R2 > 0.3. The direction of the arrow represents the correlation with changes in community composition, and the length of the arrow represents R² values. Differences between before and after administration for each group were analyzed by the Wilcoxon signed-rank test. ^#p < 0.05, ^##p < 0.01. Treatment effect was analyzed by Mann-Whitney U test. ∗p < 0.05. ASV: Amplicon Sequence Variants.

Fig. 5

Heatmap showing Spearman correlations between Bacterial Taxa and Physiological Measures of Subjects in the Control and TWK10 Groups. (A) Correlations were examined between bacterial taxa (at family levels) and exercise performance indices, and (B) total BCAA and total EAA. The degree of correlation is represented by color hue: orange represents positive correlation; blue represents negative correlation. Significant values are denoted as ∗p < 0.05, p < 0.01 and ∗∗∗p < 0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig. 6

Differences in Functional Orthologs and KEGG Pathways of the Fecal Microbial Community in the TWK10 Group. (A) Principal coordinate analysis (PCoA) of the KO profiles, (B) KEGG pathways that discriminated the fecal microbiomes and (C) Corresponding bar plot of KEGG pathway function differential analysis comparing samples from control after administration (Control_AF, n = 16) versus control before administration (Control_BE, n = 16) and TWK10 after administration (TWK10_AF, n = 16) versus TWK10 before administration (TWK10_BE, n = 16).