DOI QR코드

DOI QR Code

Evaluation of anti-inflammatory efficacy of Lacticaseibacillus rhamnosus L22-FR28 (KACC 92513P) isolated from infant feces and its Oenanthe javanica ferments

영아분변 유래 Lacticaseibacillus rhamnosus L22-FR28(KACC 92513P) 균주와 미나리 발효물의 항염증 효능 평가

  • Seoyeon Kwak (Department of Agrofood Resources, Fermented and Processed Food Science Division, National Institute of Agricultural Science, RDA) ;
  • Hee-Min Gwon (Department of Agrofood Resources, Fermented and Processed Food Science Division, National Institute of Agricultural Science, RDA) ;
  • Soo-Hwan Yeo (Department of Agrofood Resources, Fermented and Processed Food Science Division, National Institute of Agricultural Science, RDA) ;
  • So-Young Kim (Department of Agrofood Resources, Fermented and Processed Food Science Division, National Institute of Agricultural Science, RDA)
  • 곽서연 (농촌진흥청 국립농업과학원 농식품자원부 발효가공식품과) ;
  • 권희민 (농촌진흥청 국립농업과학원 농식품자원부 발효가공식품과) ;
  • 여수환 (농촌진흥청 국립농업과학원 농식품자원부 발효가공식품과) ;
  • 김소영 (농촌진흥청 국립농업과학원 농식품자원부 발효가공식품과)
  • Received : 2023.11.28
  • Accepted : 2024.02.07
  • Published : 2024.06.30

Abstract

The purposes of this study were to isolate the potential Lacticaseibacillus spp. from the feces of infants before weaning, to investigate the safety of antibiotics resistance and beta-haemolysis, and to evaluate the anti-bacterial and anti-inflammatory effects between the selected strains and Oenanthe javanica (Oj) fermented by them. As a result of analyzing the intestinal microbial community among the stools of four infants, the genus Bifidobacterium was the most dominant, but Lacticaseibacillus (L.) rhamnosus was the most frequently isolated because of the easy culture. Nine test strains, including Lactobacillus rhamnosus LGG (ATCC 53103) as the positive control, were sensitive against 8 kinds of antibiotics without vancomycin in comparison with the cut-off values at the European Food Safety Authority (EFSA), and there was no hemolysis. In the antibacterial activity experiment, the Lacticaseibacillus rhamnosus L22-FR28 (L28, KACC 92513P) strain and Oj+L28 ferment showed significantly (p<0.05) higher activities than LGG against Bacillus cereus and Staphylococcus aureus. Additionally, these decreased the activity of the NF-kB/AP-1 transcription factor and inhibited the nitric oxide and cytokines (TNF-α and IL-6) produced in macrophage RAW cells stimulated by lipopolysaccharide (LPS). Consequently, the L. rhamnosus L28 strain and Oenanthe javanica+L. rhamnosus L28 (Oj+L28) ferment selected with the high anti-inflammatory effect will improve health functionality after more research, such as the verification of animal level and identification of mechanism on an anti-inflammatory.

본 연구는 6개월 미만의 4명의 영아 분변에서 생육이 용이한 우점균을 분리하였고, 이들 중 Lactobacillus속의 유용한 균주를 발굴, 이를 이용하여 발효시켜 제조한 미나리 배양물에 대한 기능적 특성을 살펴보았다. 8주의 분리균주들 중 항생제 내성, 용혈성 등의 안전성을 확인한 후, L. rhamnosus L28과 L. reuteri 11-2 등 2주를 항염증 효능 평가를 위해 선발하였다. 선발한 시험균주 단독과 이들로 미나리를 발효시켜 제조한 미나리발효물 등의 시료들을 각각 처리하여 병원성 세균 4종에 대한 생육 억제능을 살펴본 결과, L. rhamnosus L28 균주 단독처리만으로도 양성대조구인 L. rhamnosus LGG균에 비해 B. cereus와 Sta. aureus 등에 대한 높은 항균활성을 나타내었고, 이 균주를 이용하여 발효시킨 미나리배양물(Oj+L28) 역시도 동일한 병원균들에 대해 25.03±0.00-36.14±0.00 mm의 범위로 억제환을 나타내어 높은 유해균 생육 억제능을 보였다. 또한, LPS로 자극시켜 대식세포 RAW cell에서 분비된 NF-kB/AP-1 전사인자, 활성산소(NO) 및 염증성 매개물질(TNF-α와 IL-6)의 활성 또는 생성량 억제 정도를 살펴본 결과, 양성대조구(LGG) 대비 시험균주 L. rhamnosus L28 단독 처리와 이 균으로 제조한 미나리 발효물(Oj+L28) 처리구는 항염증 효능을 나타내는 모든 지표에서 통계적 유의수준(p<0.05)으로 높은 억제 활성을 나타내었다. 특히, 사이토카인 IL-6 항목에서 L. rhamnosus L28 단독 처리의 억제율이 처리량별로 83, 96 및 99%로 L. reuteri 11-2균주보다 높은 억제 효과를 보였으며, Oj+L28 발효물 역시도 59-72%로 Oj 원액 및 Oj+LGG 배양물보다 낮은 생성량을 보였다. 또한 염증성 매개물질인 TNF-α와 IL-6의 생성량 역시도 L. rhamnosus L28 단독 처리 시 통계적 유의수준(p<0.05)으로 가장 낮은 값을 보여, LPS와 같은 항원 제시에 의해 자극된 면역세포의 기능조절에 도움을 줄 것으로 사료된다. 따라서, 본 연구에서 처음으로 발굴한 L. rhamnosus L28 균주와 이 균주로 발효시켜 제조한 미나리발효물의 높은 항염증 효과를 확인하여, 향후 동물시험 수준의 효능검증 및 기전 구명 등 심화연구를 통해 건강기능성 소재로서의 활용성을 높이고자 한다.

Keywords

Acknowledgement

This research was conducted with support from the Agricultural Science and Technology Research and Development Project of the National Institute of Agricultural Sciences, Rural Development Administration (Project Number: PJPJ01599303).

References

  1. Ahn H, Lee GS. Isorhamnetin and hyperoside derived from water dropwort inhibits inflammasome activation. Phytomedicine, 24, 77-86 (2017)
  2. Ai G, Huang ZM, Liu QC, Han YQ, Chen X. The protective effect of total phenolics from Oenanthe javanica on acute liverfailure induced by D-galactosamine. J Ethnopharmacol, 186, 53-60 (2016)
  3. Ann YG. Probiotic lactic acid bacteria. J Korean Soc Food Sci Nutr, 24, 817-832 (2011)
  4. Chae SW. Function and activation of NF-κB in immune system. Korean J Otolaryngol, 48, 284-288 (2005)
  5. Dawood MA, Koshio S, Ishikawa M, Yokoyama S, El Basuini MF. Effects of dietary supplementation of Lactobacillus rhamnosus or/and Lactococcus lactis on the growth, gut microbiota and immune responses of red sea bream, Pagrus major. Fish Shellfish Immunol, 49, 275-285 (2016)
  6. Fukushima Y, Kawata Y, Hara H, Terada A, Mitsuoka T. Effect of a probiotic formula on intestinal immunoglobulin a production in healthy children. Int J Food Microbiol, 30, 39-44 (1998)
  7. Gil NY, Choi BY, Park SY, Cho YS, Kim SY. Physicochemical properties of Doenjang using grain type Meju fermented by Aspergillus oryzae and protease. Korean J Food Preserv, 24, 697-706 (2018)
  8. Gil NY, Choi BY, Yeo SH, Kim SY. Anti-inflammatory effect of fermented Doenjang containing a halophytes Suaeda asparagoides (Miq.) powder on RAW 264.7 cells. Korean J Food Nutr, 32, 651-661 (2019)
  9. Han YQ, Huang ZM, Yang XB, Liu HZ, Wu GX. In vivo and in vitro anti-hepatitis B virus activity of total phenolicsfrom Oenanthe javanica. J. Ethnopharmacol 118, 148-153 (2008)
  10. Kang HY, Kim DH, Lee SW, Kim HC, Cho YV, Gal SW. Biological probiotic properties of Lactobacillus rhamonosus GG-4 isolated from infant feces. J Life Sci, 20, 1882-1888 (2010)
  11. Kim EJ, Song BN, Jeong DS, Kim SY, Cho YS, Park SY. Anti-oxidative and anti-inflammatory activities of fermented turmeric (Curcuma longa L.) by Rhizopus oryzae. J Life Sci 27, 1315-1323 (2017)
  12. Kim JY, Kim KH, Lee YJ, Lee SH, Park JC, Nam DH. Oenanthe javanica extract accelerates ethanol metabolism in ethanol-treated animals. BMB Rep, 42, 482-485. (2009)
  13. Kim YJ, Ko SY, Yeon YE, Lim J, Han BK, Kim H, Ahn JK, Kim D. Draft genome sequence of lytic bacteriophage CF1 infecting Citrobacter freundii isolates. Korean J Microbiology, 54, 79-80 (2018)
  14. Lee HY, Yoo MJ, Chung HJ. Antibacterial activities in watercress Oenanthe javanica D.C. Cultivated with different culture methods. Kor J Food Culture, 16, 243-249 (2001)
  15. Lee MS, Shin KS. Macrophage activation by polysaccharides from Korean's commercial and traditional soy sauces. J Korean Soc Food Sci Nutr, 26, 797-805 (2013)
  16. Makras L, Triantafyllou V, Fayol-Messaoudi D, Adriany T, Zoumpopoulou G, Tsakalidou E, Servin A, Devuy st L. Kinetic analysis of the antibacterial activity of probiotic lactobacilli towards Salmonella enterica serovar. typhimurium reveals a role for lactic acid and other inhibitory compounds. Res Microbiol, 157, 241-247 (2006)
  17. Park SJ, Lee KS, An HL. Effects of dropwort powder on the quality of castella. J East Asian Soc, Dietary Life, 17, 834-839 (2007)
  18. Seo YS, Shin KS. Immune system-stimulating activities of mucilage polysaccharides isolated from Opuntia humifusa. J Korean Soc Food Sci Nutr, 41, 95-102 (2012)
  19. Vrese M, Stegelmann A, Ritcher B, Fenselau S, Laue C, Schrezenmeir J. Probiotics: Compensation for lactase insufficiency. Am J Clin Nutr, 73, S421-S429 (2001)
  20. Zhou Q, Feng F, Yang Y, Zhao F, Du R, Zhou Z. Characterization of a dextran produced by Leuconostoc pseudomesenteroides XG5 from homemade wine. Int J Biol Macromol, 107, 2234-2241 (2018)