• 한국어업기술학회:학술대회논문집
• /
• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
• /
• pp.145-146
• /
• 2000

The presence of cross-reacting muramidase in Lactobacillus delbrueckii ssp. bulgaricus ULl2 was shown by using monoclonal antibodies raised against an muramidase-2 of Enterococcus hirae ATCC 9790. The separation of protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western immunoblot confirmed the presence of one cross-reacting band in Enterococcus hirae with an estimated molecular mass of 80 kDa, L. bulgaricus cultured cells harvested after 4 and 12 h were submitted to different autolysin releasing procedures and the liberated products were allowed to cross-react with muramidase-2 antibodies in order to estimate the efficiency of each treatment. Although the cultured cells harvested after 4 h yielded only a slight immune-reaction in Western immunoblots against these enzyme monoclonal antibodies, a strong signal was observed for the cell walls obtained from the same experimental conditions and treated with Triton X-100 surfactant. The same phenomenon was also observed by light fluorescence microscopy. Immune-labelling followed by optical and electron microscopy have shown that the muramidase-2 of L. bulgaricus ULl2 was essentially localized in the innermost part of the cell wall.(omitted)

• 생명과학회지
• /
• 제28권3호
• /
• pp.300-306
• /
• 2018

라이소자임은 선천성 면역 물질로 개불을 포함하는 여러 무척추동물의 병원균에 대한 방어에 주요하게 작용한다. 본 논문은 개불(Urechis unicinctus)의 체벽 조직 추출물로부터 무척추형 라이소자임의 정제와 그 특성에 관한 분석을 기술하고 있다. 체벽 추출물은 우선적으로 Sep-Pak C18 cartridge를 사용하여 부분적으로 분리되었으며, 분리된 분획 중 60% 메탄올에 용출된 분획이 Bacillus subtilis KCTC 1021에서 강한 항균활성을 나타내었다. 그 후 여러 단계의 역상과 이온교환 고속액체크로마토그래피(High Performance Liquid Chromatography, HPLC)를 사용하여 항균성 물질이 정제되었으며, 분자량은 약 14 kDa이었다. 이 단백질의 일차서열은 LC-MS/MS를 통해 분석되었으며, 얻은 부분적 아미노산 서열을 NCBI BLAST를 통하여 분석해 본 결과, 이 항균성 물질은 다른 동물들로부터 동정된 무척추동물형 라이소자임(invertebrate-type lysozyme)의 서열과 유사도를 가지고 있어, 체벽으로부터 정제된 개불 라이소자임(Urechis unicinctus invertebrate-type lysozyme from body wall, Uu-iLysb)으로 명명하였다. 개불 라이소자임의 활성을 확인하기 위하여 항균활성 및 라이소자임 효소 활성실험을 진행한 결과, 개불 라이소자임이 항균활성과 라이소자임 효소활성 모두 강하게 가지고 있는 것을 확인하였다.

• 대한수의학회지
• /
• 제38권4호
• /
• pp.771-776
• /
• 1998

Synthetic oligonucleotide primers of 20 and 21 bases, respectively, were used in the polymerase chain reaction (PCR) to amplify a sequence of the mrp gene, which encodes the muramidase released protein of Streptococcus suis. Amplification was not recorded when 5 other streptococcal species were tested or when 9 different nonstreptococcal species were tested. A DNA fragment of 517bp was amplified from the genomic DNA of S suis. The lower detection limit was 100pg of the genomic DNA. The primers recognized 34 serotypes of S suis reference strains and 9 isolates from pneumonic lung, brain, nasal discharge, tonsil. This results suggest that the amplification of the mrp gene by PCR method is potential for the identification of S suis isolates.

• Molecules and Cells
• /
• 제44권7호
• /
• pp.517-528
• /
• 2021

A recent genetic study with Brucella abortus revealed the secretion activator gene A (SagA) as an autolysin component creating pores in the peptidoglycan (PGN) layer for the type IV secretion system (T4SS) and peptidoglycan hydrolase inhibitor A (PhiA) as an inhibitor of SagA. In this study, we determined the crystal structures of both SagA and PhiA. Notably, the SagA structure contained a PGN fragment in a space between the N- and C-terminal domains, showing the substrate-dependent hinge motion of the domains. The purified SagA fully hydrolyzed the meso-diaminopimelic acid (DAP)-type PGN, showing a higher activity than hen egg-white lysozyme. The PhiA protein exhibiting tetrameric assembly failed to inhibit SagA activity in our experiments. Our findings provide implications for the molecular basis of the SagA-PhiA system of B. abortus. The development of inhibitors of SagA would further contribute to controlling brucellosis by attenuating the function of T4SS, the major virulence factor of Brucella.