Rakhmawatie Maya Dian, Wibawa Tri, Lisdiyanti Puspita, Pratiwi Woro Rukmi, Damayanti Ema
Doctoral Program in Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia.
Department of Biomedical Sciences, Faculty of Medicine, Universitas Muhammadiyah Semarang, Semarang 50273, Indonesia.
Iran J Basic Med Sci. 2021 Aug;24(8):1058-1068. doi: 10.22038/ijbms.2021.56468.12601.
This study explored Indonesian Actinobacteria which were isolated from endophytic microbes and mangrove ecosystem for new antimycobacterial compounds.
Antimycobacterial activity test was carried out against H37Rv. Chemical profiling of secondary metabolite using Gas Chromatography-Mass Spectroscopy (GC-MS) and High Resolution-Mass Spectroscopy (HR-MS) was done to the ethyl acetate extract of active strain InaCC A758. Molecular taxonomy analysis based on 16S rRNA gene and biosynthetic gene clusters analysis of polyketide synthase (PKS) and non-ribosomal peptide synthetase (NRPS) from InaCC A758 have been carried out. Bioassay guided isolation of ethyl acetate extract was done, then structural elucidation of active compound was performed using UV-Vis, FT-IR, and NMR spectroscopy methods.
The chemical profiling using HR-MS revealed that InaCC A758 has the potential to produce new antimycobacterial compounds. The 16S rRNA gene sequencing showed that InaCC A758 has the closest homology to strain NBRC 14599 (99.64%). In addition, InaCC A758 has NRPS gene and related to (92% of similarity), and also PKS gene related to PKS-type borrelidin of S. rochei and (74% of similarity). Two compounds with potential antimycobacterial were predicted as 1) Compound 1, similar to dimethenamid (CHClNOS; MW 275.0723), with MIC value of 100 µg/ml, and 2) Compound 2, actinomycin D (C2HNO; MW 1254.6285), with MIC value of 0.78 µg/ml.
Actinomycin D has been reported to have antimycobacterial activity, however the compound has been predicted to resemble dimethenamid had not been reported to have similar activity.
本研究探索从内生微生物和红树林生态系统中分离得到的印度尼西亚放线菌，以寻找新的抗分枝杆菌化合物。
对H37Rv进行抗分枝杆菌活性测试。使用气相色谱 - 质谱联用仪（GC-MS）和高分辨率质谱仪（HR-MS）对活性菌株InaCC A758的乙酸乙酯提取物进行次生代谢产物的化学分析。基于16S rRNA基因进行分子分类分析，并对InaCC A758的聚酮合酶（PKS）和非核糖体肽合成酶（NRPS）进行生物合成基因簇分析。对乙酸乙酯提取物进行生物测定导向分离，然后使用紫外可见光谱、傅里叶变换红外光谱和核磁共振光谱方法对活性化合物进行结构解析。
使用HR-MS进行的化学分析表明，InaCC A758有产生新的抗分枝杆菌化合物的潜力。16S rRNA基因测序显示，InaCC A758与菌株NBRC 14599的同源性最高（99.64%）。此外，InaCC A758具有NRPS基因，与（相似度92%）相关，并且还具有与罗氏链霉菌的PKS型硼雷素相关的PKS基因（相似度74%）。预测有两种具有潜在抗分枝杆菌活性的化合物，1）化合物1，类似于二甲吩草胺（CHClNOS；分子量275.0723），MIC值为100μg/ml，2）化合物2，放线菌素D（C2HNO；分子量1254.6285），MIC值为0.78μg/ml。
据报道放线菌素D具有抗分枝杆菌活性，然而，预测与二甲吩草胺相似的该化合物尚未见有类似活性的报道。
