한국식품영양학회지 Vol. 33 No. 3 (p.223-236)

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프로바이오틱스 Enterococcus faecium CKDB003의 항생제 내성 및 안전성 평가

Antibiotic Resistance and Safety Assessment of Enterococcus faecium CKDB003 for Using as Probiotics
키워드 :
probiotics,Enterococcus faecium CKDB003,antibiotic resistance,safety assessment

목차

Abstract
서 론
재료 및 방법
   1. 배지 조성
   2. 유산균 분리 및 동정
   3. 유산균 배양
   4. 항생제 내성 검사
   5. 전장유전체 분석
   6. Hyaluronidase 활성 확인
   7. Gelatinase 활성 확인
   8. 용혈성 확인
   9. Auto-aggregation 활성 분석
   10. 혈소판 제조 및 응집반응
   11. PCR법에 의한 유전자 확인
   12. 실험 통계 처리
결과 및 고찰
   1. 균주 분리 및 동정
   2. 항생제 최소 억제농도 검사에 의한 항생제 내성
   3. 전장유전체 분석을 통한 항생제 내성 유전자와 병독성유전자 평가
   4. Hyaluronidase 활성
   5. Gelatinase 활성
   6. 용혈성 확인
   7. Auto-aggregation 활성 분석
   8. 혈소판 응집 반응 확인
   9. PCR을 통한 병독성 유전자 분석
요약 및 결론
References

초록

In this study, a safety evaluation was conducted to confirm if the Enterococcus faecium CKDB003 strain obtained by selection from a mixed fermentation of fruit and milk is suitable for use as a probiotic. The MIC value for the 10 antibiotics specified in the EFSA guidance was below the acceptable cut-off value. The antibiotic resistance genes aac(6')-li, eatAv, and msr(C) exist by whole genome sequencing, but are in the chromosome and not in the plasmid, thus confirming that there is no possibility of transmission to other microorganisms. It was confirmed that cytolysin (cylA, cylB, cylI, cylL-l, cylL-s, cylM, cylR1, cylR2), aggregation substance (asa1, asp1), collagen adhesion (ace), enterococcal surface protein (esp), endocarditis antigen (efaA), hyaluronidase (hyl) and gelatinase (gelE) were not present in the genome by examining the genes of factors related to virulence. Also, the biochemical analysis showed no toxic enzyme activities, and no virulence genes were detected by the PCR method. Thus, the E. faecium CKDB003 strain can be safely used as a health functional food probiotic, based on the results of the safety assessment.