Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Development and Application of Detection Method for Aphanizomenon flos-aquae not Usable as a Food Materials in Korea

식품 중 사용금지 원료인 Aphanizomenon flos-aquae 검출법 개발 및 응용

  • Park, Yong-Chjun (New Hazardous Substance Team, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Shin, Seung-Jung (Health/Functional Food Standardization Division, Ministry of Food and Drug Safety) ;
  • Lee, Ho-Yeon (New Hazardous Substance Team, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Kim, Yong-Sang (New Hazardous Substance Team, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Kim, Mi-Ra (New Hazardous Substance Team, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Lee, Sang-Jae (New Hazardous Substance Team, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Lee, Hwa-Jung (New Hazardous Substance Team, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety)
  • 박용춘 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 신종유해물질팀) ;
  • 신승정 (식품의약품안전처 건강기능식품기준과) ;
  • 이호연 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 신종유해물질팀) ;
  • 김용상 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 신종유해물질팀) ;
  • 김미라 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 신종유해물질팀) ;
  • 이상재 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 신종유해물질팀) ;
  • 이화정 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 신종유해물질팀)
  • Received : 2013.04.05
  • Accepted : 2013.06.04
  • Published : 2013.06.30
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Abstract

Anatoxin-a, saxitoxin and neosaxitoxin are produced by Aphanizomenon flos-aquae that is a sort of the cyanobacteria phylum. Therefore, it is not permitted for food materials in Korea. Traditionally, the classification of cyanobacteria has been based on morphological characters such as trichome width, cell size, division planes, shape, and the presence of character such as gas vacuole. But, some diagnostic features, such as gas vacuole or akinetes, can show variation with different environmental or growth conditions and even be lost during cultivation. Therefore, we developed detection method for functional foods containing Aph. flos-aquae by PCR. To design the primer, 16S rRNA region of Aph. flos-aquae, Spirulina laxissima, and Spirulina spp. registered in the GeneBank (www.ncbi.nlm.nih.gov) have been used and for comparative analysis, BioEdit ver. 7.0.9.0. was used. As a result, we was design AFA-F1/AFA-R1 (363 bp) primer for the differentiation Aph. flos-aquae from chlorella, spirulina, green tea, and spinach. Also, it could be distinguished chlorella and spirulina products those are made to contain 1% Aph. flos-aquae.

Aphanizomenon flos-aquae는 시아노박테리아 일종이며 anatoxin-a, saxitoxin, neosaxitoxin 등의 독소를 생산할 수 있어 국내에서는 식품원료로 사용이 금지되어있다. 전통적으로 시아노박테리아는 사상체 넓이, 세포 크기, 분열방법, 세포형태, 가스주머니의 존재유무 등의 형태학적 특징에 의한 분류가 가능하다. 그러나 가스주머니 혹은 무성포자와 같은 특징은 주변 환경 또는 생장조건에 따라 차이가 있으며 경우에 따라 소실되기도 한다. 따라서 PCR에 의한 Aph. flos-aquae를 함유하는 기능식품을 검출할 수 있는 분석법을 개발하였다. 프라이머를 설계하기 위하여 유전자은행(www.ncbi.nlm.nih.gov)에 등록되어있는 Aph. flos-aquae, 스피루리나의 16S rRNA 염기서열을 이용하였으며, 비교 및 분석에는 BioEdit ver. 7.0.9.0 프로그램을 사용하였다. 최종적으로 클로렐라, 스피루리나, 녹차, 시금치로부터 Aph. flos-aquae를 검출할 수 있는 AFA-F1/AFA-R1(363 bp) 프라이머를 최종 선정하였다. 그리고 상기 프라이머는 Aph. flos-aquae가 각각 1% 함유 되도록 제조된 클로렐라, 스피루리나 제품에서 모두 혼입여부의 확인이 가능함을 확인하였다.

Keywords

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