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자반고등어에서 histamine 분해능을 가진 세균의 분리 동정

Isolation and Identification of a Histamine-degrading Barteria from Salted Mackerel

  • 황수정 (동의대학교 식품영양학과) ;
  • 김영만 (동의대학교 식품영양학과)
  • Hwang Su-Jung (Department of Food Science and Nutrition, Dong-eui University) ;
  • Kim Young-Man (Department of Food Science and Nutrition, Dong-eui University)
  • 발행 : 2005.10.01

초록

Histamine은 적색육 어류의 histidine이 어육 중의 Morganella morganii, Hafnia alvei 및 Klebsiella pneumoniae와 같은 부패세균에 의해 탈탄산 되어 초기에 형성되는 것으로 allergy성 식중독을 일으킬 수 있다. 이는 적색육 어류인 고등어의 선도저하 시에 많이 생성된다. 그리고 부패 후기에는 histamine을 분해하는 세균도 존재하는 것으로 알려져 있다. 그러므로, histamine 식중독의 잠재력을 지닌 자반고등어로 인한 식중독 사고 예방과 그 위생 대책을 수립하는데 필요한 자료를 얻고자 자반고등어에서 histamine 분해능을 가진 균을 분리, 동정하였다. 시료는 대형마트에서 시판되는 상태로 구입하였다. 질소원과 탄소원으로써 histamine만을 첨가한 제한배지를 사용하여 histamine 분해능을 가진 균을 분리하였다. 그리고 Cram staining, oxidase, catalase, citrate, TSI test, $H_{2}S$ reaction 및 indole 생성 등의 기본적인 생화학적 동정시험을 거쳐 10종의 시험균주를 선택하였다. 이 균주들을 16SrRNA gene 염기서열 비교에 의한 계통발생학적 분석을 이용하여 동정 하였다. 그 결과, Pseudomonas putida strain RA2, Halomonas marina, Uncultured Arctic sea ice bacterium clone ARKXV1/2-136, Halomonas venusta, Psychrobacter sp. HS5323, Pseudemonas putida KT2440, Rhodococcus erythropolis, Klebsiella terrigena (Raoultella terrigena), Alteromonadaceae bacterium T1, Shewanella massilia의 10종이 모두 동정 되 었으며, 각각 $100\%,{\;}100\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}100\%,{\;}95\%,{\;}99\%,{\;}100\%$의 상동성을 보였다. Histamine분해능의 존재를 탁도측정법과 효소법에 의해 확인한 결과, 분리된 10종 모두의 histamine 분해능이 재확인 되었고, 그 중 Shewanella massilia가 최대의 histamine 분해능을 보이는 것으로 확인되었다. 이 결과로 자반고등어 시판 제품에는 다수의 histamine 분해 세균이 존재하는 것을 확인할 수 있었으며, 이 세균을 활용한다면 식품 내 존재하는 histamine을 효과적으로 분해할 수 있을 것이라 예상된다.

Histamine can be produced at early spoilage stage through decarboxylation of histidine in red-flesh fish by Proteus morganii, Hafnia alvei or Klebsiella pneumoniae. Allergic food poisoning is resulted from the histamine produced when the freshness of Mackerel degrades. Conversely it has been reported that there are bacteria which decompose histamine at the later stage. We isolated histamine decomposers from salted mackerel and studied the characteristics to help establish hygienic measure to prevent outbreak of salted mackerel food poisoning. All the samples were purchased through local supermarket. Histamine decomposers were isolated using restriction medium using histamine 10 species were selected. Identification of these isolates were carried out by the comparison of 16S rDNA partial sequence; as a result, we identified Pseudomonas putida strain RA2 and Halomonas marina, Uncultured Arctic sea ice bacterium clone ARKXV1/2-136, Halomonas venusta, Psychrobacter sp. HS5323, Pseudomonas putida KT2440, Rhodococcus erythropolis, Klebsiella terrigena (Raoultella terrigena), Alteromonadaceae bacterium T1, Shewanella massilia with homology of $100\%,{\;}100\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}100\%,{\;}95\%,{\;}99\%,{\;}and{\;}100\%$respectively. Turbidometry determination method and enzymic method were employed to determine the ability of histamine decomposition. Among those species Shewanella massilia showed the highest in ability of histamine decomposition. From these results we confirmed various histamine decomposer were present in salted mackerel product in the market.

키워드

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