초록 ▼

Ⅳ. 연구개발결과
1. 황색포도알균 중 식중독 세균인 황색포도상구균과 리스테리아균 제어에 효과적인 박테리오신 생산균주인 Staphylococcus pasteuri RSP-1을 선발하고 전체 게놈 정보를 분석하여 Gene bank에 등록하였다.
2. 소수성 레진을 박테리오신을 분리하고 열처리 후 RP-HPLC를 이용하여 박테리오신 단백질을 순수분리, 농축하여 고 활성, 고 순도의 박테리오신 분리기법을 확립하였다.
3. 분리한 박테리오신의 농작물에서 식중독균 억제 효과를 검증하였다.
4. 황색포도상구균을 제어하는

Ⅳ. 연구개발결과
1. 황색포도알균 중 식중독 세균인 황색포도상구균과 리스테리아균 제어에 효과적인 박테리오신 생산균주인 Staphylococcus pasteuri RSP-1을 선발하고 전체 게놈 정보를 분석하여 Gene bank에 등록하였다.
2. 소수성 레진을 박테리오신을 분리하고 열처리 후 RP-HPLC를 이용하여 박테리오신 단백질을 순수분리, 농축하여 고 활성, 고 순도의 박테리오신 분리기법을 확립하였다.
3. 분리한 박테리오신의 농작물에서 식중독균 억제 효과를 검증하였다.
4. 황색포도상구균을 제어하는 박테리오파지를 분리하는 효과적인 박테리오파지 분리법을 확립하고 새로운 박테리오파지 6종을 분리하였다.
5. 파지의 용균 관련 유전자를 분리하고 발현을 통하여 식중독균의 병원성에 미치 는 영향에 대해 검증하였다.
6. 박테리오파지 SP5와 SP6에서 endolysin을 분리, 클로닝 및 과발현하여 여러 식중독균에 대한 lytic activity spectrum을 조사하고 분석하였다.

Abstract ▼

1. Bacteriocin
Staphylococcus aureus is recognized as a globally important pathogen in food-borne disease and have rates of antimicrobial resistance. Therefore, attention has been focused on Bacteriocins that are considered safe and effective as food additives to prevent food pathogens. In this s

1. Bacteriocin
Staphylococcus aureus is recognized as a globally important pathogen in food-borne disease and have rates of antimicrobial resistance. Therefore, attention has been focused on Bacteriocins that are considered safe and effective as food additives to prevent food pathogens. In this study describes the purification, characterization a novel bacteriocin produced by Staphylococcus pasteuri RSP-1. The supernatant of S. pasteuri RSP-1 possessed antimicrobial activity against Gram(+) bacteria and strong activity against S. aureus. Purification of the active compound was achieved using a combination of precipitation with XAD-16, dialysis, and reverse-phase high-performance liquid chromatography(RP-HPLC). The molecular mass of purified bacteriocin was 1.5 kDa determined by Matrix-assisted laser desorption ionization-time of flight(MALDI-TOF). Purified bacteriocin was stable in a pH range between 1.0 and 10.0 at 121 °C for 30 min, and showed resistance to α-amylase, lipase, and RNase, and DNase. However, its activity was lost after trypsin, pronase E, pepsin, α-chymotrypsin, proteinase K, and papin treatment. The results suggest that the possibility of using novel bacteriocin for application in prevention/treatment of food containing agricultural products or pharmaceutical industries.
2. Bacteriophage
As methicillin-resistant S. aureus (MRSA) has become increasingly prevalent, controlling this pathogen with standard antibiotic treatment has become challenging. Bacteriophages (phages) and endolysin have attracted interest as alternative antibacterial agents to control MRSA. We isolated a lot of S. aureus phages from hands and farm environments such as animal feces and soils. Based on the results of host range determination with 150 S. aureus strains and restriction enzyme treatment of phage DNA, two phages, designated SP5 and SP6, were selected for further characterization and genome sequencing. Both SP5 and SP6 were classified as members of the family Siphoviridae. The genome of SP5 comprises 43,305 bp and contains 63 ORFs, while the SP6 genome comprises 42,902 bp and contains 61 ORFs. Although they have different host spectra, the phage genomes exhibit high nucleotide similarity to each other. Adsorption assay results suggested that the host range determinants of the two phages are involved in both adsorption and infection. Comparative genomic analyses of the two phages provided evidence that the lysogenic/lytic control module and tail proteins may be important for host specificity. As well as bacteriophage, endolysin also have been considered as an attracted biocontrol agent. Bioinformatic analysis revealed putative lytic protein, SP6_endolysin, which had high homology with N-acetylmuramoyl_L-alanine amidase. The SP6_endolysin was purified and its bacteriolytic activity was demonstrated. Taken together, S. aureus-targeting bacteriophages and their endolysins have high potential as biocontrol agents for S. aureus.