• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.511-520
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• 2015

Triclosan, the widely used biocide, specifically targets enoyl-acyl carrier protein reductase (ENR) in the bacterial fatty acid synthesis system. Although the fish pathogen Aeromonas salmonicida subsp. salmonicida exhibits triclosan resistance, the nature of this resistance has not been elucidated. Here, we aimed to characterize the triclosan resistance of A. salmonicida subsp. salmonicida causing furunculosis. The fosmid library of triclosan-resistant A. salmonicida subsp. salmonicida was constructed to select a fosmid clone showing triclosan resistance. With the fosmid clone showing triclosan resistance, a subsequent secondary library search resulted in the selection of subclone pTSR-1. DNA sequence analysis of pTSR-1 revealed the presence of a chromosomal-borne fabV-encoding ENR homolog. The ENR of A. salmonicida (FabVas) exhibited significant homology with previously known FabV, including the catalytic domain YX₍₈₎K. fabVas introduction into E. coli dramatically increased its resistance to triclosan. Heterologous expression of FabVas might functionally replace the triclosan-sensitive FabI in vivo to confer E. coli with triclosan resistance. A genome-wide search for fabVas homologs revealed the presence of an additional fabV gene (fabVas2) paralog in A. salmonicida strains and the fabVas orthologs from other gram-negative fish pathogens. Both of the potential FabV ENRs expressed similarly with or without triclosan supplement. This is the first report about the presence of two potential FabV ENRs in a single pathogenic bacterium. Our result suggests that triclosan-resistant ENRs are widely distributed in various bacteria in nature, and the wide use of this biocide can spread these triclosan-tolerant ENRs among fish pathogens and other pathogenic bacteria.

• Natural Product Sciences
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• 제28권2호
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• pp.45-52
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• 2022

The identification of Plasmodium falciparum enoyl acyl-carrier protein reductase (pfENR) is considered as a potential biological target against malaria. Trema orientalis is considered a rich source of phytochemicals useful in malaria treatment. This study evaluated the in-vitro inhibitory activity of the extract and isolated compounds of T. orientalis leaf; the isolated compounds and the analogues of the most active compound were subjected to in-silico molecular docking studies on pfENR. The methanolic extract of T. orientalis was subjected to repeated chromatographic separation which led to the isolation of some compounds. The isolated compounds from the plant were examined for their antimalarial activity using β-hematin inhibition assay. Virtual screening via molecular docking and ADMET studies were conducted to gain insight into the mechanism of binding of ligand and to identify effective pfENR inhibitors. The isolated compounds and the analogues of the most active isolates were gotten from PubChem library for use in docking study. Hexacosanol and β-sitosterol showed inhibition of the β-hematin formation. The docking results showed that hexacosanol, β-sitosterol and the analogues of β-sitosterol displayed binding energy ranging between -6.1 kcal/mol and -11.6 kcal/mol. Sitosterol glucoside has the highest docking score. Some of the ligands showed more binding affinity than known bioactive compounds used as reference. Analogues of β-sitosterol has been shown to be potential inhibitors of pfENR, therefore, the findings from this study suggest that sitosterol glucoside and ergosterol peroxide could act as antimalarial agents after further lead optimisation investigations.

• 생명과학회지
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• 제26권4호
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• pp.460-467
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• 2016

사상체 토착 남세균을 경상남도 합천군 합천호의 수화시료로부터 무균적으로 분리하였으며, 형태적·분자적 동정 결과 림노트릭스 속에 속하는 것으로 밝혀졌다. 따라서, 본 남세균 균주는 림노트릭스 속 KNUA012 균주로 명명하였으며, 분리균주의 최적생장 온도는 섭씨 25도였다. 지질성분 분석 결과, 에스테르 교환반응을 거치지 않고 직접 연료로 사용할 수 있는 펜타데칸(C₁₅H₃₂)과 헵타데칸(C₁₇H₃₆)과 같은 알칸들이 본 균주에 의해 광독립 영양적으로 생합성 된다는 것이 밝혀졌다. 또한 알칸 생합성에 관여하는 유전자들이 본 남세균 내에 존재하는 것을 발견하였다. 일반적인 미세조류 바이오디젤 구성성분으로 알려진 미리스트올레산(C_14:1), 팔미트산(C_16:0) 및 팔미톨레산(C_16:1) 역시 KNUA012 균주에 의해 주요 지방산 성분으로서 생산되는 것으로 확인되었다. 근사분석 결과 KNUA012 균주의 휘발성물질 함량은 86.0%였으며, 원소분석 결과 고위발열량은 19.8 MJ kg⁻¹으로 나타났다. 또한, 본 분리균주는 고부가가치 항산화물질로 알려져 있는 피코시아닌을 광독립영양적으로 21.4 mg g⁻¹의 농도로 생산할 수 있는 것을 확인하였다. 따라서, 본 연구결과는 KNUA012 균주가 미세조류 기반 바이오연료와 바이오매스 원료의 경제적인 생산에 있어 이상적인 자원이 될 수 있음을 제시하였다.