Abstract
여러 원천에서 분리한 유산균을 대상으로 신규 bacteriocin 생산 유산균을 선발하고 API 당 발효성과 16S rDNA를 분석한 결과 P. damnosus 로 동정되어 JNU 534로 명명하였다. P. damnosus JNU 534에 의해 생산된 bacteriocin은 다수의 유산균과 L. monocytogenes의 생육을 억제하였지만, Gram음성균을 포함한 다른 병원성균들은 저항성을 나타냈다. Bacteriocin의 생산은 대수기 초반에 시작되어 정지기 초기에 최대 활성에 이르렀다. Bacteriocin은 pH 2-9의 넓은 범위와 <TEX>$100^{\circ}C$</TEX>에서 15분간의 열처리에 대해서도 안정성을 유지하였다. 또한 각종 단백질 분해효소에 의해 활성을 상실함으로써 본 연구에서 정제한 항균 물질이 단백질성 물질임을 확인할 수 있었으며 30% ammonium sulfate 침전과 <TEX>$C_{18}$</TEX> chromatography를 통하여 bacteriocin을 정제 할 수 있었다. Tricine-SDS-PAGE에 의해 bacteriocin의 분자량을 약 3.4 kDa으로 추정하였으며, 지시균의 생육 저지환의 위치가 bacteriocin band와 일치하였다. 정제된 bacteriocin으로부터 분석한 N-terminal 아미노산 부분 서열은 <TEX>$NH_2$</TEX>-ILLEELNV로 나타났다. A new bacteriocin-producing lactic acid bacteria (LAB) which has been isolated from kimchi was identified as Pediococcus damnosus by use of API kit and 16S rDNA sequencing, and designated as P. damnosus JNU 534. The bacteriocin produced by P. damnosus JNU 534 markedly inhibited the growth of some of LAB and Listeria monocytogenes, whereas other pathogens including Gram negative bacteria were not susceptible. The production of bacteriocin started at the beginning of exponential phase and reached maximum activity at the early stationary phase. The bacteriocin was stable on the wide pH range of 2-9 and heat treatment up to <TEX>$100^{\circ}C$</TEX> for 15 min. The antimicrobial compound was inactivated by treatments of proteolytic enzymes indicating its proteinaceous in nature. The bacteriocin was purified by 30% ammonium sulfate precipitation followed by hydrophobic interaction column and <TEX>$C_{18}$</TEX> column chromatography. The estimated molecular weight of the bacteriocin using tricine SDS-PAGE was approximately 3.4 kDa and the identified N-terminal amino acid sequence was <TEX>$NH_2$</TEX>-ILLEELNV.
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