Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Isolation and Characteristics of Microorganisms Producing Extracellular Enzymes from Jeju Traditional Fermented Soybean Paste (Doenjang)

제주전통된장으로부터 세포외효소 분비능이 우수한 미생물의 분리 및 특성

  • Oh, You-Sung (Biotechnology Regional Innovation Center, Jeju National University) ;
  • Park, Ji-Eun (Biotechnology Regional Innovation Center, Jeju National University) ;
  • Oh, Hyun-Jeong (Biotechnology Regional Innovation Center, Jeju National University) ;
  • Kim, Jung-Hyon (Dept. of Tourism & Food Service Cuisine, Cheju Tourism College) ;
  • Oh, Myung-Cheol (Dept. of Tourism Hotel Culinary Arts, Jeju College of Technology) ;
  • Oh, Chang-Kyung (Dept. of Tourism Hotel Culinary Arts, Jeju College of Technology) ;
  • Oh, Young-Ju (Dept. of Hotel Culinary Arts, Jeju Halla College) ;
  • Lim, Sang-Bin (Biotechnology Regional Innovation Center, Jeju National University)
  • 오유성 (제주대학교 생명과학기술혁신센터) ;
  • 박지은 (제주대학교 생명과학기술혁신센터) ;
  • 오현정 (제주대학교 생명과학기술혁신센터) ;
  • 김정현 (제주관광대학 관광외식조리계열) ;
  • 오명철 (제주산업정보대학 관광호텔조리과) ;
  • 오창경 (제주산업정보대학 관광호텔조리과) ;
  • 오영주 (제주한라대학 호텔조리과) ;
  • 임상빈 (제주대학교 생명과학기술혁신센터)
  • Published : 2010.01.30
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Abstract

Bacteria strains with high activities of extracellular enzymes (protease, fibrinolytic enzyme, amylase, cellulase, and lipase) were isolated from Jeju traditional fermented soybean paste (Doenjang), and characterized by 16S rRNA gene sequence analysis and physiological properties. Protease activities were higher in JR14, JR19, JR25, JR32, JR38, JR47, and JR64 than Bacillus subtilis KCCM 12027 (standard strain). Amylase activities were shown in JR6, JR25, JR38, JR56 and JR81, while not in KCCM12027. Cellulase activities were higher in JR6, JR14, JR48, and JR65 than those of other isolated strains and KCCM 12027 whereas lipase activities were the higher in JR-14 and JR-48. Thrombolytic activity in JR19 with high hemolysis activity were 192% compared with that of plasmin as a positive control. Zymogram analysis indicated that the thrombolytic active strains had 4~5 bands in the molecular weight range of 25~75 kDa. Gene sequence analysis of rRNA revealed that the isolated stains had 99% homology with Bacillus species, and the thrombolytic active stain JR19 was B. stratosphericus $41KF2a^T$.

제주 전통된장으로부터 세포외효소(protease, fibrinolytic enzyme, amylase, cellulase, lipase) 분비능이 우수한 세균을 분리한 후 16S rRNA 유전자 분석과 생리적 특성을 분석하여 균주를 확인하고자 하였다. Protease 분비능은 JR14, JR19, JR25, JR32, JR38, JR47과 JR64가 표준균주인 Bacillus subtilis KCCM12027보다 활성이 높았다. Amylase 분비능은 JR6, JR25, JR38, JR56, JR81에서 나타난 반면 표준균주인 KCCM12027에서는 나타나지 않았다. Cellulase 분비능은 JR6, JR14, JR48과 JR65가 다른 분리균주 또는 표준균주보다 높았으며, lipase 분비능은 JR14와 JR48이 높았다. 혈전용해 활성은 positive control인 plasmin의 용해 영역에 비하여 JR19가 192%로 가장 높았고, hemolysis 활성도 높았다. 혈전용해능이 있는 균주인 JR19, JR32, JR47, JR64의 배양액을 zymography한 결과, 25～75 kDa 사이에서 4～5개의 밴드가 확인되었다. 된장으로부터 분리한 균주들의 16s rRNA 유전자 염기서열을 분석한 결과 모두 Bacillus species와 99%의 상동성을 나타내었으며, 혈전용해능이 가장 우수한 JR19는 B. stratosphericus $41KF2a^T$와 거의 일치하였다.

Keywords

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