[논문]한국형 프로바이오틱스에 의한 쓴메밀 내 rutin의 생물전환

권창; 김종원; 박영광; 강승범; 정명준; 김수정; 임상현

doi:10.48022/mbl.2302.02004

한국형 프로바이오틱스에 의한 쓴메밀 내 rutin의 생물전환
Bioconversion of Rutin in Tartary Buckwheat by the Korean Indigenous Probiotics 원문보기

Microbiology and biotechnology letters = 한국미생물·생명공학회지, v.51 no.1, 2023년, pp.83 - 92

권창 ((주)쎌바이오텍 세포공학연구소) , 김종원 ((주)쎌바이오텍 세포공학연구소) , 박영광 ((주)쎌바이오텍 세포공학연구소) , 강승범 ((주)쎌바이오텍 세포공학연구소) , 정명준 ((주)쎌바이오텍 세포공학연구소) , 김수정 (농촌진흥청 국립식량과학원 고령지농업연구소) , 임상현 ((주)쎌바이오텍 세포공학연구소)

초록
AI-Helper

본 연구에서는 한국형 프로바이오틱스와 볶은 쓴메밀을 배양하여 쓴메밀의 대표 성분인 rutin에서 quercetin으로 생물전환을 확인하였다. 프로바이오틱스 17종의 유전체 분석 결과 CBT BG7, LC5, LR5, LP3, LA1과 LGA1 등 6종에서 rutin에서 isoquercetin 전환에 관련된 α-rhamnosidase 유전자가 확인되었다. Isoquercetin에서 quercetin 전환에 관련된 β-glucosidase 유전자는 17종의 프로바이오틱스에서 모두 확인되었다. 17종의 프로바이오틱스 중 CBT BG7, LR5, LP3, LA1, LGA1과 ST3 등 6종의 균주가 배양 7일 후 rutin에서 quercetin으로 최대 21.5 ± 0.3%까지 생물전환 하였다. 생물전환 시간 단축을 위하여 프로바이오틱스와 복합 효소 Cellulase KN^®을 함께 배양하였다. 그 결과, 효소 첨가 없이 가장 높은 생물전환율 21.5 ± 0.3%를 보였던 CBT LA1 균주는 효소와 함께 배양 1일 후 84.6 ± 0.5%의 생물전환율을 보였다. 특히 CBT ST3 (96.2 ± 0.4%), SL6 (90.1 ± 1.4%) 그리고 LP3 (90.0 ± 0.4%) 균주는 quercetin으로 90% 이상의 높은 전환율을 보였다. 추가로 프로바이오틱스 생물전환능이 우수하여 quercetin 함량이 높은 배양여액 동결건조물은 RAW264.7 세포에서 LPS에 의해 유도된 NO 생성 억제 효과를 보였다. 본 연구를 통해 한국형 프로바이오틱스와 효소를 조합하여 볶은 쓴메밀과 함께 배양하였을 때 rutin으로부터 항염증, 항산화, 항비만, 항당뇨 활성을 가지는 생리활성 물질 quercetin으로 생물전환이 증가하고 전환 시간이 단축됨을 확인하였다.

Abstract ▼ AI-Helper

In this study, bioconversion of rutin to quercetin was confirmed by the fermentation of Korean indigenous probiotics and tartary buckwheat. Based on whole genome sequencing of 17 probiotics species, α-rhamnosidase, related to bioconversion of isoquercetin (quercetin 3-β-D glucoside) from rutin, is identified in the genome of CBT BG7, LC5, LR5, LP3, LA1, and LGA1. β-Glucosidase, related to bioconversion of isoquercetin to quercetin, is identified in the genome of all 17 species. Among the 17 probiotics species, 6 probiotics including CBT BG7, LR5, LP3, LA1, LGA1 and ST3 performed the bioconversion of rutin to quercetin up to 21.5 ± 0.3% at 7 days after fermentation. The fermentation of each probiotics together with enzyme complex Cellulase KN® was conducted to reduce the time of bioconversion. As a result, CBT LA1 which showed the highest yield of bioconversion of 21.5 ± 0.3% when the enzyme complex was not added showed high bioconversion yield of 84.6 ± 0.5% with adding the enzyme complex at 1 day after fermentation. In particular, CBT ST3 (96.2 ± 0.4%), SL6 (90.1 ± 1.4%) and LP3 (90.0 ± 0.4%) showed high yield of bioconversion more than 90%. In addition, such probiotics including high levels in quercetin indicated the inhibitory effects of NO production in LPS-induced RAW264.7 cells. In this study, we confirmed that the fermentation of Korean indigenous probiotics and enzyme complex together with roasted tartary buckwheat increased the content of quercetin and reduced the time of bioconversion of rutin to quercetin which is a bioactive compound related to anti-inflammatory, antioxidants, anti-obesity, and anti-diabetes.

주제어

표/그림 (9)

표 Table 1. Probiotics species used in this study and their origin.
그림 Fig. 1. Cultivation of tartary buckwheat Fagopyrum tataricum L. Gaertn cv. Hwangguem-miso (A), seed of tartary buckwheat (B) and roasted seed of tartary buckwheat (C).
표 Table 2. CBT probiotics species encoding α-rhamnosidase and β-glucosidase.
그림 Fig. 2. Concentrations of rutin, isoquercetin and quercetin at 7 days after the fermentation with CBT probiotics and roasted tartary buckwheat.
표 Table 3. The yield of bioconversion of rutin and isoquercetin to quercetin according to the incubation time of CBT BG7, LR5, LP3, LA1, LGA1 and ST3. Quercetin Yield (mol queretin/mol rutin + isoquercetin)
그림 Fig. 3. Concentrations of rutin, isoquercetin and quercetin at 1 day after the fermentation with CBT probiotics, enzyme complex and roasted tartary buckwheat.
표 Table 4. The yield of bioconversion of rutin and isoquercetin to quercetin with each probiotics and enzyme complex. Quercetin Yield (mol queretin/mol rutin + isoquercetin)
그림 Fig. 4. HPLC analysis of rutin, isoquercetin and quercetin in the media fermented by the probiotics and/or enzyme complex with roasted tartary buckwheat. (A) Standards, (B) Media fermented by CBT LA1 for 7d, (C) Media fermented by CBT LA1 and enzyme complex Cellulase KN® for 1d, (D) Media fermented by CBT LP3 for 7d, and (E) Media fermented by CBT LP3 and enzyme complex Cellulase KN® for 1d.
그림 Fig. 5. The effect of 8 probiotics with high bioconversion ratio on the cell viability (A) and nitrate production (B) in RAW264.7 cells. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with control.

참고문헌 (32)

Kim SJ, Sohn HB, Suh JT, Kim GH, Hong SY, Kim KD, et al. 2017a.？Domestic and overseas status of buckwheat production and？future trends. J. Korean Soc. Int. Agr. 29: 226-233.
Kim SJ, Sohn HB, Hong SY, Lee JN, Kim KD, Suh JT, et al. 2020.？Construction of data system on seed morphological traits and？functional component in tartary buckwheat germplasms.？Korean J. Plant Res. 33: 446-459.
Kim SJ, Kim YH. Agricultural guide Buckwheat. 2018. Rural？Development Administration, Jeonju, Korea. pp. 07-90.
Park CH. 2006. Development of new cultivar and technology of？cultivation and utilization in tartary buckwheat as a new agricultural material. pp. 12-197. Ministry of Agriculture.
Ikeda K. 2002. Buckwheat composition, chemistry, and processing. Adv. Food Nutr. Res. 44: 395-434.

상세보기
Kang HW. 2014. Antioxidant and anti-inflammation effects of？water extract from buckwheat. Korean J. Culinary Res. 20: 190-199.
Kim SH, Lee EY, Ham SS. 2007. Antioxidation and antigenotoxic？effects of buckwheat sprout extracts. J. Korean Soc. Food Sci.？Nutr. 36: 955-959.
Lee CC, Hsu WH, Shen SR, Cheng YH, Wu SC. 2012. Fagopyrum？tataricum (buckwheat) improved high-glucose-induced insulin？resistance in mouse hepatocytes and diabetes in fructose-rich？diet induced mice. Exp. Diabetes Res. 2012: 375673.

상세보기
Kim DW, Hwang IK, Lim SS, Yoo KY, Li H, Kim YS, et al. 2009.？Germinated buckwheat extract decreases blood pressure and？nitrotyrosine immunoreactivity in aortic endothelial cells in？spontaneously hypertensive rats. Phytother. Res. 23: 993-998.
Chang KJ, Seo GS, Kim YS, Huang DS, Park JI, Park JJ, et al. 2010.？Components and biological effects of fermented extract from？tartary buckwheat sprouts. Korean J. Plant Res. 23: 131-137.
Yoon BR, Cho BJ, Lee HK, Kim DJ, Rhee SK, Hong HD, et al. 2012.？Antioxidant and anti-adipogenic effects of ethanolic extracts？from tartary and common buckwheats. Korean J. Food Preserv.？19: 123-130.
Nam HK, Hong SH, Shin KC, Oh DK. 2012. Quercetin production？from rutin by a thermostable β-rutinosidase from Pyrococcus？furiosus. Biotechnol. Lett. 34: 483-489.
Kim JA. 2006. Study on the anti-obesity effects of germinatedbuckwheat. Ms theses, Kangwon National University, Chuncheon, Korea.
Fukushima T, Tokuji Y, Kinoshita M, Ohnishi M, Kawahara M,？Ohba K. 2008. Anti-inflammatory effect of buckwheat sprouts？in lipopolysaccharide-activated human colon cancer cells and？mice. Biosci. Biotechol. Biochem. 72: 3148-3157.
Holasova M, Fiedlerava V, Smrcinova H, Orsak M, Lachman J,？Vavreinova S. 2002. Buckwheat-the source of antioxidant activity in functional foods. Food Res. Int. 35: 207-211.

상세보기
Kim SJ, Sohn HB, Lee KT, Shin JS, Kim S, Nam JH, et al. 2019.？Anti-inflammatory effects of seed ethanolic extracts of the？common buckwheat and tartary buckwheat are mediated？through the suppression of inducible nitric oxide synthase and？pro-inflammatory cytokines in LPS-induced RAW 264.7 macrophage cells. Korean J. Food Sci. Technol. 51: 565-575.
Erlund I. 2004. Review of the flavonoids quercetin, hesperetin,？and naringenin. Dietary sources, bioactivities, bioavailability？and epidemiology. Nutr. Res. 24: 851-874.

상세보기
Zhang ZL, Zhou ML, Tang Y, Li FL, Tang YX, Shao JR. et al. 2012.？Bioactive compounds in functional buckwheat food. Food Res.？Int. 49: 389-395.

상세보기
Kim JH, Lee S, Cho EJ, Kim HY. 2019. Neuroprotective effects of？kaempferol, quercetin, and its glycosides by regulation of？apoptosis. J. Korea Acad. Ind. Coop. Soc. 20: 286-293.
Heim KE, Tagliaferro AR, Bobilya DJ. 2002. Flavonoid antioxidants: Chemistry, metabolism and structureactivity relationships. J. Nutr. Biochem. 13: 572-584.

상세보기
Ademosun AO, Oboh G, Bello F, Ayeni PO. 2015. Antioxidative？properties and effect of quercetin and its glycosylated form？(Rutin) on acetylcholinesterase and butyrylcholinesterase？activities. J. Evid. Based Complementary Altern. Med 21: NP11-7.
Yang J, Lee H, Sung J, Kim Y, Jeong HS, Lee J. 2019. Conversion？of rutin to quercetin by acid treatment in relation to biological？activities. Prev. Nutr. Food Sci. 24: 313-320.

상세보기
Kim GN, Kwon YI, Jang HD. 2011. Protective mechanism of？quercetin and rutin on 2,2'-azobis(2-amidinopropane) dihydrochloride or Cu 2+ -induced oxidative stress in HepG2 cells.？Toxicol. in Vitro 25: 138-144.

상세보기
Salehi B, Machin L, Monzote L, Sharifi-Rad J, Ezzat SM, Sale MA,？et al. 2020. Therapeutic potential of quercetin: New insights？and perspectives for human health. ACS Omega 5: 11849-11872.

상세보기
Zuo Y, Xin X, Xu H, Yuan H, Li X, Yu Y, et al. 2020. Highly efficient？bioconversion of flavonoid glycosides from citrus processing？wastes in solvent-buffer systems. Green Chem. 22: 3196-3207.

상세보기
Dewi RT, Ekapratiwi Y, Sundowo A, Ariani N, Yolanda T, Filaila E.？2019. Bioconversion of quercetin glucosides from Dendrophthoe？pentandra leaf using Aspergillus acueletus LS04-3. AIP Conference Proceedings 2175: 020048.
Huynh NT, Smagghe G, Gonzales GB, Camp JV, Raes K. 2018.？Bioconversion of kaempferol and quercetin glucosides from？plant sources using Rhizopus spp. Fermentation 4: 1-9.
Amaretti A, Raimondi S, Leonardi A, Quartieri A, Rossi M. 2015.？Hydrolysis of the rutinose-conjugates flavonoids rutin and？hesperidin by the gut microbiota and bifidobacteria. Nutrients？7: 2788-2800.

상세보기
Choi J, Kwon C, Kim JW, Chung MJ, Yoon JH, Lim S. 2022. Bioconversion of Ginsenosides by Bifidobacterium CBT BG7, BR3？and BL3. Microbiol. Biotechnol. Lett. 50: 395-403.
Kim SJ, Sohn HB, Nam JH, Lee JN, Chang DC, Kim YH. 2022.？Comparison of rutin content and quality characteristics of tea？products from common buckwheat (Fagopyrum esculentum)？and tartary buckwheat (Fagopyrum tataricum) by different processing and brewing methods. Korean J. Food Sci. Technol. 54:？185-195.
Park CM, Kim GM, Cha GS. 2021. Biotransformation of flavonoids？by newly isolated and characterized Lactobacillus pentosus？NGI01 strain from Kimchi. Microorganisms 9: 1075.

상세보기
Mueller M, Zartl B, Schleritzko A, Stenzl M, Viernstein H, Unger？FM. 2018. Rhamnosidase activity of selected probiotics and？their ability to hydrolyse flavonoid rhamnoglucosides, Bioprocess？Biosyst. Eng. 41: 221-228.？

저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증