Low-fat Yogurt Supplemented with Novel Probiotic Lactobacillus brevis KU200019 and

Fructooligosaccharide Showed Immunomodulatory Activity and Attenuated Oxidative Stress

Menaka Menike Kariyawasam Majuwana Gamage1, Yeon Jin Kim1, Na-Kyoung Lee1, and Hyun-Dong Paik1
+82-10-9767-4049
1Department of Food Science and Biotechnology of Animal Resources, Konkuk University, Seoul 05029, Republic of Korea masha6440@gmail.com
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Abstract Results and Discussion
This study was performed to develop a functional synbiotic  Scavenging ROS in living cells  Immunomodulatory activity
yoghurt enriched with Lactobacillus brevis KU200019 and
fructooligosaccharides (FOS). The yoghurt samples were (A) (B)

manufactured using a commercial starter culture and labeled as

C (a control without probiotics and FOS), P (L. brevis ATCC
14869), K (L. brevis KU200019), PS (L. brevis ATCC 14869
and FOS), and KS (L. brevis KU200019 and FOS). The
samples were stored at 4 ± 1 °C for 21 days. The KS sample
observed with high reactive oxygen species scavenging (2.52-
fold), and immunomodulatory activities. Supplementation of
probiotic or FOS did not have an adverse effect on sensory
properties. These results confirmed that L. brevis KU200019 (C) (D)
and FOS could be used in the dairy industry to improve the
putative health benefits and quality of the dairy products.
Materials and Methods (A)

 Bacterial strains and culture conditions

Lactobacillus brevis KU200019 and
Lactobacillus brevis ATCC 14869
 Yoghurt sample preparation
 Control – Yoghurt with starter culture (C ) (E)

 Sample 1 – Yoghurt with L. brevis ATCC 14869 (P)

 Sample 2 – Yoghurt with L. brevis KU200019 (K)
 Sample 3 – Yoghurt with L. brevis ATCC 14869 and FOS (PS)
 Sample 4 – Yoghurt with L. brevis KU200019 and FOS (KS)

 Scavenging ROS in living cells Figure 1. Fluorescence images of RAW 264.7 macrophages.
Negative control without LPS, positive control 10 ng/mL of LPS,
 ROS scavenging activity by DCF-DA C;10 ng/mL of LPS and 0.5 mg of protein/mL of the WSE from
Morphological changes by fluorescence microscope sample C, P; 10 ng/mL of LPS and 0.5 mg of protein/mL of the WSE
from sample P, K; 10 ng/mL of LPS and 0.5 mg of protein/mL of the
 Immunomodulatory activity WSE from sample K, PS; 10 ng/mL of LPS and 0.5 mg of
 NO assay protein/mL of the WSE from sample PS, and KS 10 ng/mL of LPS Figure 2. Immunomodulatory effects of WSE from yoghurt samples. (A) nitric oxide
and 0.5 mg of protein/mL of the WSE from sample KS, (A) (NO), (B) iNOS, (C) TNF-α, (D) IL-6, and (E) IL-1β production. RAW 264.7 macrophages
 Real time PCR correspondence fold changes in ROS expression. Different letters on treated with 10 ng/mL of LPS were used as the positive control and cells incubated in the
 Organoleptic properties evaluation the top of the bars are significantly different (p<0.05). All values are medium without WSE or LPS were used as the negative control. The quantification results are
mean of three replicates (mean±SD). expressed relative to the control (untreated) after being normalized to β-actin. Different letters
 Semi-trained panelist (30 members) on the top of the bars are significantly different (p<0.05). All values are mean of three
 5 point hedonic scale Conclusions replicates (mean±SD).

The results demonstrate that synbiotic yoghurt has high ROS activity. The WSE stimulated the release of NO and up-regulated the gene expression of iNOS and pro-inflammatory cytokines at the mRNA level
(TNF-α, IL-1β, and IL-6) in RAW 264.7 cells. In addition, supplementing the yoghurt with probiotic or FOS did not have adverse effect on sensorial properties. These results confirmed that L. brevis KU200019 and
FOS have potential use in the fermented dairy industry as a functional adjunct culture, to improve the putative health benefits and quality of the product.