치태형성을 억제하는 유산균 K1270을 김치로부터 분리한 후 배양적, 생화학적 특징 및 16S rDNA염기서열 분석에 의해 Pediococcus pentosaceus K1270으로 동정하였다. K1270 균주는 5% 자당이 함유된 배지에서 Streptococcus mutans Ingbritt에 의한 인공치태 형성을 대조군에 비해 94.6 % 억제하였으며, 비수용성 글루캔의 합성을 89.6 % 억제하였다. S. mutans Ingbritt의 증식은 대조군에 비해 100배 정도 억제하였다. K1270균주는 TMB와 peroxidase가 첨가된 MRS 한천배지에서 과산화수소를 생산하였으며, 인공치태 형성에 대한 K1270균주의 억제 효과는 catalase 첨가에 의해 일부 감소되었다. K1270 균주의 배양 상청액을 10% 자당이 함유된 $2{\times}M17$ broth에 동량 가한 경우 인공치태 형성 및 Streptococcus mutans Ingbritt의 증식이 억제되었으며, 이 억제효과는 catalase첨가에 의해 일부 감소되었고, 열처리, 또는 trypsin 처치에 의해 완전히 소실되었다. 따라서, 본 연구에서 분리, 동정된 P. pentosaceus K1270은 과산화 수소와 bacteriocin 유사물질을 분비하여 S. mutans Ingbritt의 증식을 억제함으로써 인공치태 형성을 억제하는 것으로 사료된다.
치태형성을 억제하는 유산균 K1270을 김치로부터 분리한 후 배양적, 생화학적 특징 및 16S rDNA염기서열 분석에 의해 Pediococcus pentosaceus K1270으로 동정하였다. K1270 균주는 5% 자당이 함유된 배지에서 Streptococcus mutans Ingbritt에 의한 인공치태 형성을 대조군에 비해 94.6 % 억제하였으며, 비수용성 글루캔의 합성을 89.6 % 억제하였다. S. mutans Ingbritt의 증식은 대조군에 비해 100배 정도 억제하였다. K1270균주는 TMB와 peroxidase가 첨가된 MRS 한천배지에서 과산화수소를 생산하였으며, 인공치태 형성에 대한 K1270균주의 억제 효과는 catalase 첨가에 의해 일부 감소되었다. K1270 균주의 배양 상청액을 10% 자당이 함유된 $2{\times}M17$ broth에 동량 가한 경우 인공치태 형성 및 Streptococcus mutans Ingbritt의 증식이 억제되었으며, 이 억제효과는 catalase첨가에 의해 일부 감소되었고, 열처리, 또는 trypsin 처치에 의해 완전히 소실되었다. 따라서, 본 연구에서 분리, 동정된 P. pentosaceus K1270은 과산화 수소와 bacteriocin 유사물질을 분비하여 S. mutans Ingbritt의 증식을 억제함으로써 인공치태 형성을 억제하는 것으로 사료된다.
Pediococcus pentosaceus K1270 was isolated from naturally fermented kimchi and identified based on the 16S rDNA sequence as well as cultural and biochemical characteristics. This strain strongly inhibited the formation of biofilm by Streptococcus mutans Ingbritt. K1270 also showed antibacterial acti...
Pediococcus pentosaceus K1270 was isolated from naturally fermented kimchi and identified based on the 16S rDNA sequence as well as cultural and biochemical characteristics. This strain strongly inhibited the formation of biofilm by Streptococcus mutans Ingbritt. K1270 also showed antibacterial activity against S. mutans Ingbritt. It was observed that K1270 strain produced hydrogen peroxide on MRS agar supplemented with 3, 3, 5, 5-tetramethylbenzidine (TMB) and peroxidase and the inhibitory effect of K1270 strain on the biofilm formation was reversed by the addition of catalase in part. Culture supernatant of K1270 inhibited the biofilm formation and the multiplication of S. mutans Ingbritt. This inhibitory effect of culture supernatant was decreased slightly by the addition of catalase and abolished by heat or trypsin treatment. Thus, this study suggests that P. pentosaceus K1270 inhibit the biofilm formation through the inhibition of the replication of S. mutans Ingbritt by producing hydrogen peroxide and bacteriocin.
Pediococcus pentosaceus K1270 was isolated from naturally fermented kimchi and identified based on the 16S rDNA sequence as well as cultural and biochemical characteristics. This strain strongly inhibited the formation of biofilm by Streptococcus mutans Ingbritt. K1270 also showed antibacterial activity against S. mutans Ingbritt. It was observed that K1270 strain produced hydrogen peroxide on MRS agar supplemented with 3, 3, 5, 5-tetramethylbenzidine (TMB) and peroxidase and the inhibitory effect of K1270 strain on the biofilm formation was reversed by the addition of catalase in part. Culture supernatant of K1270 inhibited the biofilm formation and the multiplication of S. mutans Ingbritt. This inhibitory effect of culture supernatant was decreased slightly by the addition of catalase and abolished by heat or trypsin treatment. Thus, this study suggests that P. pentosaceus K1270 inhibit the biofilm formation through the inhibition of the replication of S. mutans Ingbritt by producing hydrogen peroxide and bacteriocin.
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제안 방법
In this study, we have isolated and identified the LAB, which inhibited the biofilm formation by S. mu- tans Ingbritt, from kimchi, Korean traditional vegetable food that many LAB inhabit. Furthermore, the inhibitory mechanism of action was also investigated.
The amplified PCR products were recovered and purified using a Wizard PCR Preps DNA Purification System (Promega). The purified PCR products were sequenced with an automatic DNA sequencer (ABI 310, Perkin-Elmer) using the primer 5'- AGAGTTTGATCMTGGCTCAG-3, (nucleotide positions 8 to 27 of the Escherichia coli 16S rRNA gene).
H2O2 production was determined by observing the color change of K1270 strain into blue. To find whether inhibiting substance against biofilm formation was H2O2, catalase(10, 000 U or 20, 000 U) was added into culture mixture of S. mutans Ingbritt and K1270, and then beaker-wire test was carried out. The biofilm on wires were weighed after 24 h incubation at 3713.
성능/효과
42%) to the Pediococcus pentosaceus sequence. Based on the 16S rDNA sequence as well as cultural and biochemical characteristics, the isolated K1270 was identified as Pediococcus pentosaceus and named Pediococcus pentosaceus K1270.
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