DOI QR코드

DOI QR Code

Analysis of Enhancement Effect and Attachment Ability of Beneficial Intestinal Microflora in Puffed Grain Foods Using Confocal Laser Scanning Microscopy

곡물 소재 팽화식품에서 장내 유익균의 증진 효과 분석 및 공초점 현미경을 이용한 부착능 평가

  • 정명교 (충북대학교 식품생명.축산과학부) ;
  • 오도건 (충북대학교 식품생명.축산과학부) ;
  • 권오성 ((주)지디앤와이) ;
  • 정준영 ((주)지디앤와이) ;
  • 이임식 (식품의약품안전처) ;
  • 김광엽 (충북대학교 식품생명.축산과학부)
  • Received : 2017.06.13
  • Accepted : 2017.07.10
  • Published : 2017.09.30

Abstract

This study examined the adhesiveness of beneficial intestinal bacteria to whole-grains using confocal scanning laser microscopy (CLSM), to demonstrate the prebiotic effects of whole-grains, and to develop prebiotic puffed snacks with these whole-grains. CLSM has been used to observe the adhesiveness of Lactobacillus acidophilus, which belongs to beneficial intestinal bacteria, to whole-grain powders using optical sectioning techniques. The enhanced effects on the growth of beneficial intestinal bacteria with the hot water grain extract were verified using an indirect count method. Finally, a puffed snack was produced with the prebiotic effect and the quality was evaluated by checking the chromaticity and degree of hardness. As a result, L. acidophilus exhibited adhesive ability to whole-grain powders and growth of selected beneficial intestinal bacteria were improved significantly. The Hunter L value of the developed puffed snack increased when seasoning was added. The hardness of the puffed snack with seasoning was higher than that of the control. The results of a sensory evaluation showed that the puffed snack with seasoning was highly rated in the overall preference compared to the control.

본 연구에서는 공초점 주사현미경(CLSM)을 활용하여 장내 유익균과 국산 통곡물(whole grain) 소재들과의 부착을 통해 생육환경에 있어 도움이 될 수 있는 서식공간을 제공받는 것을 시각화하고, 소재들의 프리바이오틱스 효과를 in vitro 배양을 통해 확인하여 통곡물 소재들로 제조된 프리바이오틱스 팽화식품을 개발하는 것을 목표하였고, 그 결과 장 속에서 장내 유익균 균총을 향상시켜 인체의 건강증진에 유익한 팽화과자 제조에 필요한 기초 자료를 확보하였다. 선정곡물들의 일반성분 함량은 수분이 8.99~12.19%, 조회분 1.34~2.39%, 조단백질 7.34~15.38%, 조지방 1.84~8.12%로 함량이 분석되었고, 불용성 식이섬유 3.08~18.62%, 수용성 식이섬유 0.10~3.13%, 그리고 총 식이섬유 함량 5.26~21.36%로 확인되었다. 곡물분말의 프리바이오틱스 부착여부를 CLSM으로 관찰한 결과 곡물분말과 혼합된 Lactobacillus acidophilus는 곳곳에 산재되어 있었지만, 주변 유동액 층보다 상대적으로 곡물분말 주변에 부착되어 존재하였다. 선정곡물들의 장내 유익균의 증진에 활성 효과가 있음을 검증한 결과 L. acidophilus에서는 찰보리 추출물에서 가장 높은 흡광도 값을 보였다. Lactobacillus delbrueckii subsp. lactis에서는 귀리 추출물에서 가장 높은 효과를 보였다. Streptococcus salivarius subsp. thermophilus에서는 control보다 높은 수치를 보였지만, 두드러진 차이는 보이지 않았다. Bifidobacterium bifidum은 12시간 이후 흡광도 값은 일정하게 유지되었다. 곡물 소재 동결건조 추출물 간의 장내 유익균 활성 정도를 비교한 실험 결과 L. acidophilus에서는 찰보리와 율무에서 가장 높은 생육증진 효과를 보였고 현미를 제외한 다른 곡물 모두 유의한 증진 효과를 보였다. L. delbrueckii subsp. lactis에서는 율무에서 가장 높은 증진 효과를 보였고, S. salivarius subsp. thermophilus에서는 찰보리와 찰수수에서 가장 높은 증진 효과를 보였다. 그리고 B. bifidum에서는 찰보리와 찰기장에서 높은 증진 효과를 보였다. 팽화식품 제조 시 배합에 사용될 6종의 혼합곡물의 장내 유익균 활성 효과를 알아본 결과, 24시간 배양 시 L. acidophilus는 control에 비해 1.72배이상 상승하였고, L. delbrueckii subsp. lactis는 1.89배, S. salivarius subsp. thermophilus는 1.66배 이상, B. bifidum는 1.47배가량 상승한 것으로 분석되었다. 제조된 팽화과자의 색도 측정결과 L값은 치즈맛의 시즈닝 첨가군이 대조군보다 높게 나왔다. a값은 대조군이 적색에 가깝게, 시즈닝 첨가군은 녹색 쪽으로 나타났다. b값은 두 제품에서 크게 유의한 차이를 보이진 않았다. 팽화과자의 경도는 첨가군이 대조군보다 높았고, 바삭거림은 첨가군이 대조군보다 약한 것으로 확인되었다.

Keywords

References

  1. Gibson GR, Roberfroid MB. 1995. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr 125: 1401-1412.
  2. Mitsuoka T. 1982. Recent trends in research on intestinal flora. Biosci Microflora 1: 3-24.
  3. Mitsuoka T. 1990. Bifidobacteria and their role in human health. J Ind Microbiol 6: 263-267. https://doi.org/10.1007/BF01575871
  4. Isolauri E, Salminen S, Ouwehand AC. 2004. Probiotics. Best Pract Res Clin Gastroenterol 18: 299-313. https://doi.org/10.1016/j.bpg.2003.10.006
  5. Lee KA, Kim MS, Cho HB. 2008. Effect of extract of fermented dropwort on intestinal bacteria and enzymes in vitro. Korean J Microbiol 44: 358-361.
  6. Cho NJ, Lee SK, Kim SK, Joo HK. 1998. Effect of wheat flour brew with Bifidobacterium bifidum on rheological properties of wheat flour dough. Korean J Food Sci Technol 30: 832-841.
  7. Lee HA, Lee SS, Shin HK. 1997. Effect of apple dietary fiber on the in vitro growth of intestinal bacteria. Korean J Food Sci Technol 29: 107-114.
  8. Park JM, Lee JH, Hong SI. 2014. A research trend analysis of probiotics and prebiotics. Food Science and Industry 47: 54-66.
  9. Cocolin L, Alessandria V, Dolci P, Gorra R, Rantsiou K. 2013. Culture independent methods to assess the diversity and dynamics of microbiota during food fermentation. Int J Food Microbiol 167: 29-43. https://doi.org/10.1016/j.ijfoodmicro.2013.05.008
  10. Del Chierico F, Vernocchi P, Bonizzi L, Carsetti R, Castellazzi AM, Dallapiccola B, de Vos W, Guerzoni ME, Manco M, Marseglia GL, Muraca M, Roncada P, Salvatori G, Signore F, Urbani A, Putignani L. 2012. Early-life gut microbiota under physiological and pathological conditions: The central role of combined meta-omics-based approaches. J Proteomics 75: 4580-4587. https://doi.org/10.1016/j.jprot.2012.02.018
  11. Park YS, Song SK. 1998. Effect of polymer coating on initial microorganism attachment and the biofilm growth. Korean J Env Hlth Soc 24: 104-109.
  12. Kim YH, Park SO, Han KS, Oh SJ, You SK, Kim SH. 2004. Adhesion ability and inhibition of enterohemorrhagic E. coli O157:H7 adhesion to intestinal epithelial cells in Lactobacillus acidophilus. Korean J Food Sci Ani Resour 24: 86-91.
  13. Shin HK, Shin OH, Koo YJ. 1992. Effects of potato protein on the growth of Clostridium perfringens and other intestinal microorganisms. Korean J Appl Microbiol Biotechnol 20: 249-256.
  14. Biagi E, Candela M, Turroni S, Garagnani P, Franceschi C, Brigidi P. 2013. Ageing and gut microbes: Perspectives for health maintenance and longevity. Pharmacol Res 69: 11-20. https://doi.org/10.1016/j.phrs.2012.10.005
  15. Fukushima Y, Kawata Y, Hara H, Terada A, Mitsuoka T. 1998. Effect of a probiotic formula on intestinal immunoglobulin A production in healthy children. Int J Food Microbiol 42: 39-44. https://doi.org/10.1016/S0168-1605(98)00056-7
  16. Makras L, De Vuyst L. 2006. The in vitro inhibition of Gram-negative pathogenic bacteria by bifidobacteria is caused by the production of organic acids. Int Dairy J 16: 1049-1057. https://doi.org/10.1016/j.idairyj.2005.09.006
  17. Inglin RC, Stevens MJA, Meile L, Lacroix C, Meile L. 2015. High-throughput screening assays for antibacterial and antifungal activities of Lactobacillus species. J Microbiol Methods 114: 26-29. https://doi.org/10.1016/j.mimet.2015.04.011
  18. Jung SH. 2006. Application of high-speed slit scanning confocal laser microscope in biology. Abstract No TP-VI3 presented at summer Meeting of the Institute of Optical Society of Korea. Yonsei University, Seoul, Korea.
  19. Jeong SH, Cho NS. 2002. Study on morphology and physical properties of Indian mallow (Abutilon avicennae Gaertner) fibers by CLSM (I). J Korea TAPPI 34: 61-66.
  20. Kim SH, Park JM, Kim CH. 1999. Effects of fiber wall thickness on paper properties using CLSM. J Korea TAPPI 31: 39-45.
  21. Bae HC, Renchinkhand G, Ku JH, Nam MS. 2011. Characterization of fermented milk added with green whole grains of barley, wheat, glutinous rice and common rice powders. CNU J Agric Sci 38: 485-491.
  22. Ham H, Woo KS, Lee B, Park JY, Sim EY, Kim BJ, Lee C, Kim SJ, Kim WH, Lee J, Lee YY. 2015. Antioxidant compounds and activities of methanolic extracts from oat cultivars. J Korean Soc Food Sci Nutr 44: 1660-1665. https://doi.org/10.3746/jkfn.2015.44.11.1660
  23. Wood PJ, Anderson JW, Braaten JT, Cave NA, Scott FW, Vachon C. 1989. Physiological effects of beta-D-glucan rich fraction from oats. Cereal Foods World 34: 878-882.
  24. Lim SY. 2008. Inhibitory effects of methanol extracts from Korean Orysa sartiva and Coix lachryma-jobi var. ma-yuen on mutagenicity and growth of human cancer cells. J Life Sci 18: 1415-1419. https://doi.org/10.5352/JLS.2008.18.10.1415
  25. Ko JY, Song SB, Lee JS, Kang JR, Seo MC, Oh BG, Kwak DY, Nam MH, Jeong HS, Woo KS. 2011. Changes in chemical components of foxtail millet, proso millet, and sorghum with germination. J Korean Soc Food Sci Nutr 40: 1128-1135. https://doi.org/10.3746/jkfn.2011.40.8.1128
  26. Brennan MA, Derbyshire E, Tiwari BK, Brennan CS. 2013. Integration of ${\beta}$-glucan fibre rich fractions from barley and mushrooms to form healthy extruded snacks. Plant Foods Hum Nutr 68: 78-82. https://doi.org/10.1007/s11130-012-0330-0
  27. Kang MY, Sung YM. 2000. Varietal differences in quality characteristics of Yukwa (fried rice cookie) made from fourteen glutinous rice cultivars. Korean J Food Sci Technol 32: 69-74.
  28. AOAC. 2000. Official method of analysis. 17th ed. Association of Official Analytical Chemists, Arlington, VA, USA. Method 925.09, 923.03, 950.09, 963.15, 991.43.
  29. Hwang JB, Yang MO, Shin HK. 1997. Survey for approximate composition and mineral content of medicinal herbs. Korean J Food Sci Technol 29: 671-679.
  30. Korean Diabetes Association. 2012. Diabetes fact sheet in Korea 2012. Korea Centers for Disease Control and Prevention, Chungju, Korea.
  31. Grundy S. 2002. Third report of the National Cholesterol Education Program (NCEP) expert panel on detection, evaluation, and treatment of high blood cholesterol in adults (Adult Treatment Panel III) final report. Circulation 106:3143-3421.
  32. Reiner Z, Catapano AL, De Backer G, Graham I, Taskinen MR, Wiklund O, Agewall S, Alegria E, Chapman MJ, Durrington P, Erdine S, Halcox J, Hobbs R, Kjekshus J, Filardi PP, Riccardi G, Storey RF, Wood D; ESC Committee for Practice Guidelines (CPG) 2008-2010 and 2010-2012 Committees. 2011. ESC/EAS Guidelines for the management of dyslipidaemias: the Task Force for the management of dyslipidaemias of the European Society of Cardiology (ESC) and the European Atherosclerosis Society (EAS). Eur Heart J 32: 1769-1818.
  33. Jeong SY, Park MJ, Lee SY. 2011. Quality characteristics of brown rice Jeung-pyun. Korean J Food Cult 26: 86-93.
  34. Kim SH, Kim DJ. 2004. Analysis of nitrite oxidizing bacteria in a nitrite oxidizing biofilm by fluorescence in situ hybridization. Spring Meeting of the Korean Society for Biotechnology and Bioengineering. Daegu, Korea. p 120-124.
  35. Thomas CJ, McMeekin TA. 1984. Effect of water uptake by poultry tissues on contamination by bacteria during immersion in bacterial suspensions. J Food Prot 47: 398-402. https://doi.org/10.4315/0362-028X-47.5.398
  36. Dickson JS, Koohmaraie M. 1989. Cell surface charge characteristics and their relationship to bacterial attachment to meat surfaces. Appl Environ Microbiol 55: 832-836.
  37. Jang KI, Chung DH, Ha SD, Kim KS, Lee KH, Kim MG, Kim CH, Kim KY. 2006. Assessment of inactivation for Salmonella spp. on chicken meat using confocal laser microscopy and flow cytometry. Korean J Food Sci Technol 38: 290-294.
  38. Sung SK, Rhee YK, Cho CW, Kim YC, Lee OH, Hong HD. 2013. Physicochemical properties and antioxidative activity of fermented Rhodiola sachalinensis and Korean red ginseng mixture by Lactobacillus acidophilus. Korean J Food Nutr 26: 358-365. https://doi.org/10.9799/ksfan.2013.26.3.358
  39. Choi JJ, Park YR, Chung CH. 2014. Quality characteristics of snack with added Dongchimi powder. J East Asian Soc Diet Life 24: 819-826.
  40. Bae HS, Lee YK, Kim SD. 2002. Quality characteristics of Yukwa with citrus peel powder. J East Asian Soc Diet Life 12: 388-396.