Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Isolation of Bacillus subtilis GS-2 Producing γ-PGA from Ghungkukjang Bean Paste and Identification of γ-PGA

청국장으로부터 분리한 Poly(γ-glutamic acid)를 생산하는 균주 Bacillus subtilis GS-2의 분리 및 γ-PGA의 확인

  • Bang, Byung-Ho (Department of Food and nutrition Science, Eulji University) ;
  • Jeong, Eun-Ja (Department of Food and nutrition Science, Eulji University) ;
  • Rhee, Moon-Soo (Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Yong-Min (Department of Microbial Engineering, Konkuk University) ;
  • Yi, Dong-Heui (Department of Microbial Engineering, Konkuk University)
  • 방병호 (을지대학교 식품영양학과) ;
  • 정은자 (을지대학교 식품영양학과) ;
  • 이문수 (한국생명공학연구원) ;
  • 김용민 (건국대학교 미생물공학과) ;
  • 이동희 (건국대학교 미생물공학과)
  • Received : 2010.11.19
  • Accepted : 2011.03.08
  • Published : 2011.03.31
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Abstract

${\gamma}$-PGA(poly-${\gamma}$-glutamic acid) is an unusual anionic polypeptide that is made of D- and L-glutamic acid units connected by amide linkages between ${\alpha}$-amino and ${\gamma}$-carboxylic acid groups. ${\gamma}$-PGA has been isolated from many kinds of organisms. Many Bacillus strains produce ${\gamma}$-PGA as a capsular material of an extracellular viscous material. It is safe for eating as a viscosity element of fermented soybean products such as Chungkookjang and Natto. It is biodegradable, edible and nontoxic toward humans and the environment and its molecular weight varies from ten thousand to several hundred thousand depending on the kinds of strains used. Therefore, potential applications of ${\gamma}$-PGA and its derivatives have been of interest in the past few years in a broad range of industrial fields such as food, cosmetics, medicine, water-treatment, etc. In this study, a bacterium, Bacillus subtilis GS-2 isolated from the Korean traditional seasoning food, Chungkookjang could produce a large amount of ${\gamma}$-PGA with high productivity and had a simple nutrient requirement. Based on carbon utilization pattern and partial 16S rRNA sequence analysis, the GS-2 strain was identified as B. subtilis. The determination of purified ${\gamma}$-PGA was confirmed with thin layer chromatography (TLC), high performance liquid chromatography (HPLC), fourier transform infrared (FT-IR) spectra, and $^1H$-nuclear magnetic resonance ($^1H$-NMR) spectroscopy.

Poly-${\gamma}$-glutamic acid (${\gamma}$-PGA)는 청국장이 발효할 때 생성되는 점질성 물질의 하나로, monomer glutamic acid의 ${\alpha}$-아미노기와 ${\gamma}$-카르복실기 사이의 amide linkage에 의해 결합된 D(-)와 L(-) glutamic acid repeat units로 이루어진 homopolymer이다. 주로 Bacillus sp.에 의해 생산된다. ${\gamma}$-PGA는 수용성, 음이온성, 무독성, 생분해성, 생체적 합성, 식용 등의 다양한 특성을 가지고 있기 때문에 여러 분야에서 응용되고 있다. 본 실험에서 ${\gamma}$-PGA를 생산하는 균주를 우리나라의 전통발효식품인 청국장으로부터 분리하였다. 분리균주의 형태 및 배양학적, 생리학적 특성, API kit 및 16S rRNA 서열을 사용하여 동정한 결과, 분리균주 GS-2는 B. subtilis와 가장 유사하여 B. subtilis GS-2로 명명하였다. 분리 정제된 ${\gamma}$-PGA의 동정은 TLC, HPLC, FTIR 그리고 $^1H$-NMR spectroscopy를 통하여 확인하였다.

Keywords

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