Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Determination of Siderophore from Bacillus Mojavensis Using Liquid Chromatography quadrupole Time-of-flight Tandem Mass Spectrometry

액체크로마토그래피-사중극 비행시간형 탠덤질량분석기를 이용한 Bacillus mojavensis 균주 속 사이드로포어 규명

  • Cheon, Hae In (College of Pharmacy, Kyungsung University) ;
  • Yeo, Mi Seon (College of Pharmacy, Kyungsung University) ;
  • Kim, Kang Min (Department of Pharmaceutical Science and Technology, Kyungsung University) ;
  • Kang, Jae Seon (College of Pharmacy, Kyungsung University) ;
  • Pyo, Jaesung (College of Pharmacy, Kyungsung University)
  • Received : 2018.08.07
  • Accepted : 2018.11.03
  • Published : 2019.02.28
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Abstract

Recently, it has been reported that Bacillus mojavensis possesses antifungal properties and plant growth-promoting activities, which are similar to the characteristics of siderophore. In this study, the siderophore produced by B. mojavensis was assessed using a solid phase extraction (SPE) cartridge and liquid chromatography quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS). After B. mojavensis was incubated in phenol medium for 16 hr and lyophilized, the sample was dissolved in water and loaded to an SPE cartridge to remove interferences. The cartridge was washed with 5% methanol in water and eluted with 2% formic acid in methanol sequentially. The eluted solution was evaporated under a stream of nitrogen gas and reconstituted in methanol. The reconstituted sample was filtered, and $1{\mu}l$ of the sample was assessed using Q-TOF MS/MS. The mass spectrometer was operated using the positive electrospray ionization mode. Based on the mass spectrum and tandem mass spectrum, the siderophore produced by B. mojavensis was bacillibactin, one of the catechol types of siderophore with a molecular weight of 882.2556. This siderophore analysis could provide a justification for the study of B. mojavensis as a functional food and for pharmaceutical applications.

Bacillus mojavensis를 이용한 항진균성 및 식물 생장 촉진 활성이 최근 보고되었다. 이런 활성은 사이드로포어의 일반적 특성과 일치하여, 본 연구에서는 Bacillus mojavensis가 생산하는 사이드로포어를 고체상추출 카트리지와 액체크로마토그래피-사중극 비행시간형 탠덤 질량분석기를 이용하여 규명하였다. Bacillus mojavensis를 페놀 배지에서 16시간 동안 배양하고 동결 건조 시킨 후, 물에 용해시켜 고상추출 카트리지에 로딩하였다. 카트리지는 5% 메탄올로 세척하고 2% 포름산을 이용해 용출 시켰다. 용출액은 메탄올에 재용해 후 분석을 하였다. Bacillus mojavensis의 사이드로포어는 질량 스펙트럼의 결과를 바탕으로 882.2556의 분자량을 갖는 카테콜타입의 사이드로포어 중 하나인 bacillibactin으로 확인되었다. 이 사이드로포어 분석은 Bacillus 연구 및 기능성 식품 그리고 Bacillus mojavensis의 약학 응용 분야에 큰 기여를 할 것으로 예상된다.

Keywords

SMGHBM_2019_v29n2_198_f0001.png 이미지

Fig. 1. (A) The extracted ion chromatogram (EIC) as m/z 883.27 ion in positive ESI mode. (B) The mass spectral peaks of detection and extraction in 5.112 min. (C) Product ion mass spectra of sample obtained by Q-TOF-MS/MS in m/z 20-900.

Table 1. LC-Q TOF-MS/MS condition

SMGHBM_2019_v29n2_198_t0001.png 이미지

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