전통발효식품으로부터 Chitin 분해 미생물의 분리 및 특성 규명 = Isolation and Characterization of Chitinoloytic Strain, Bacillus atrophaeus CJ-3.

국문 초록 (Abstract)

전통발효 식품인 간장으로부터 chitinase활성이 높으면서 갈변반응을 일으키는 균주인 CJ-3를 분리하여 갈변반응생성물이 면역기능, 항산화력 및 세포활성에 미치는 영향과 생화학적 특성을 규명하였다. 분리된 CJ-3 균주는 16S rDNA 염기서열분석에 의하여 Bacillus atrophaeus로 동정되었으며 분리된 균주의 분자량은 대략 31.0 kDa으로 colloidal chitin(0.5, 1.0, 2.0%)의 첨가에 의하여 chitinase 활성이 현저히 증가되었다 B. atrophaeus CJ-3에 의해서 유도되는 갈변반응물 200${\mu}\ell$ 첨가로 LPS에 의하여 유도되는 nitric oxide (NO) 활성을 45%까지 감소시켰다. MTT 분석을 통한 세포활성에서도 갈변반응물은 LPS에 의하여 세포활성을 정상수준으로 회복하였다. CJ-3. 균주의 DPPH 전자공여능법에 의한 항산화력은 갈변반응에 의하여 약 55% 증가하였다 . 이러한 결과들은 갈변반응생성물은 면역기능 및 세포활성을 증가시키는 것으로 추정된다. 분리된 균주의 최적 생육조건은 최적온도 37$^{\circ}C$, 최적 pH 7.0 및 염 농도는 9% NaCl농도까지는 잘 생육하는 것으로 나타났으며 분리된 균주의 주요 intracellular 유리아미노산은 glutamate로 나타났다.

다국어 초록 (Multilingual Abstract)

A bacterial strain CJ-3 which produced chitinase was isolated from Korean traditional soy sauce. Using 16S rDNA analysis, the strain CJ-3 was identified as Bacillus atrophaeus. The approximate molecular weight of the putative chitinase enzyme was 31.0 kDa and the enzyme activity was remarkably induced by addition of colloidal chitin (0.5, 1.0, 2.0%). The antioxidant activity was increased 53% by the browning reaction products of B. atrophneus CJ-3. Escherichia. coli lipopolysaccharides (LPS)-induced production of nitric oxide(NO) was reduced up to 45% by the browning reaction product in RAW264.7 macrophage. Inhibition of cell viability in the presence of LPS was recovered to normal level by the browning reaction product. These results suggest that browning reaction of B. atrophaeus CJ-3 plays an important role for activation of immune system. B. atrophaeus CJ-3 exhibited optimum temperature and pH of 37$^{\circ}C$ and pH 7.0∼8.0, respectively. The major intracelluar free amino acid was determined to be glutamate.

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