Effect of fermentation by Lactobacillus reuteri and Lactobacillus acidophilus on antioxidant activity of quinoa extract

Fermented foods, probiotic, prebiotics, and symbiotic, are among the most important groups of functional food that have attracted the attention of researchers during the last years. Proteolytic activity of lactic acid bacteria can lead to the production of peptides in the fermented product. The produced peptides can exhibit different biological activities such as antioxidant, antihypertensive, etc. that are influenced by the type of protein source, type of bacteria, time and conditions of fermentation process. Fermentation of various cereals such as quinoa seeds with high sugar and protein content by lactic acid bacteria can lead to the production of antioxidant peptides and improving their nutritional properties.  In this study, the role of Lactobacillus reuteri and Lactobacillus acidophilus and the combination of two bacteria on the progress of fermentation and antioxidant activity of quinoa extract was investigated. The fermentation process was started by separate and simultaneous inoculating of Lactobacillus reuteri and Lactobacillus acidophilus and continued for 72 hours at 37° C. Sampling was performed every 24 hours of fermentation and samples were kept at -20° C for further analysis. The parameters such as pH, acidity, amount of soluble protein, degree of hydrolysis, amount of phenolic compounds and DPPH and ABTS radical scavenging activity were determined.  Lactobacillus acidophilus showed higher acidification capacity than Lactobacillus reuteri. The amount of acidity in the sample fermented by Lactobacillus acidophilus increased from 0.27 to 1.13 % after 72 hours, while this amount was measured as 0.80 % for sample fermented by Lactobacillus reuteri. The amount of soluble protein and the degree of hydrolysis increased in samples fermented by both species. However, the largest increase was related to the sample fermented by Lactobacillus reuteri, so that the amount of soluble protein increased from 0.72 to 0.88 mg / ml and the value of free amino groups increased from 20.28 to 58.14 µM leucin/ mg protein during 72 hours of fermentation. The DPPH and ABTS radical scavenging activity increased in all fermented samples. The highest antioxidant activity was observed in samples fermented by Lactobacillus reuteri, followed by a combination of two bacteria (50:50) and Lactobacillus acidophilus. The amounts of phenolic compounds increased in all fermented samples. However, the highest increase was related to the sample fermented by Lactobacillus reuteri, so that it increased from 0/73 to 16.21 mg Gallic acid / ml after 72 hours of fermentation. Therefore, the results showed that despite the higher acidifying power of Lactobacillus acidophilus in quinoa extract, but Lactobacillus reuteri exhibited higher proteolytic activity, more ability to produce antioxidant peptides and also release phenolic compounds during the fermentation process.Simultaneous use of the two bacteria did not intensify the proteolytic activity and antioxidant activity of peptides, and the greatest increase in acidity, proteolysis, and antioxidant activity occurred in the first 24 hours of fermentation. Fermented extracts showed higher ABTS radical inhibitory activity than DPPH radical inhibition, indicating the hydrophilic nature of most produced antioxidant compounds. The highest levels of antioxidant activity were observed in samples fermented by Lactobacillus reuteri, a combination of Lactobacillus reuteri and Lactobacillus acidophilus (50:50) and Lactobacillus acidophilus, respectively. The results showed that fermentation by Lactobacillus reutri has the greatest effect on the production of antioxidant peptides and the release of phenolic compounds. The results of this study confirm the effectiveness of fermentation methods on improving the healing properties of quinoa extract and Lactobacillus reuteri was a more effective bacterium in fermentation and production of antioxidant peptides compared to Lactobacillus acidophilus. Simultaneous use of two bacteria did not increase the intensity of fermentation and did not improve the antioxidant activity compared to single use of each bacteria. Finally, the results of this study showed that fermentation of quinoa extract improves its antioxidant properties and has the potential to be used as a fermented beverage.