Polypeptide Antibiotic Produced by a Thermo-Halophilic Bacterium from Pria Laot Sabang 76 Isolate

Jurnal Riset Kimia Pub Date : 2024-04-23 DOI:10.25077/jrk.v15i1.634

Febriani Febriani, M. A. Suzanni, Ellenia Rakashiwi, Nurdin, T. Iqbalsyah

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Abstract

Extremophiles are known to produce antibiotics. This study aimed to isolate and purify polypeptide antibiotics from Pria Laot Sabang (PLS) 76, a bacterium isolated from an undersea fumarole. The antibiotic production was done by cultivating PLS 76 in a 2.5% TSB liquid medium and incubated at 70°C, 150 rpm for 112 hours. Ninhydrin assay on the crude supernatant after centrifugation indicated that the PLS 76 produced a polypeptide antibiotic. The supernatant was partitioned using methanol. The antibiotic activity was tested using the Kirby-Bauer method, and the inhibition zone was about 12 mm against Escherichia coli and Staphylococcus aureus. The methanol fraction was purified further by Preparative Thin Layer Chromatography and produced several fractions. Fractions with the same Rf were pooled and subjected to the ninhydrin assay. Fraction A, which gave a positive result after the ninhydrin assay, showed an inhibition zone of only 7 mm against E. coli and S. aureus. The Quadrupole Time-of-Flight (QTOF) Mass Spectrometry data indicated that the antibiotic's structure was similar to Polymyxin B2, an antibiotic of a polypeptide group.

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来自 Pria Laot Sabang 76 分离物的嗜热嗜热细菌产生的多肽抗生素

众所周知，嗜极细菌能产生抗生素。本研究旨在从从海底火口分离出来的细菌 Pria Laot Sabang (PLS) 76 中分离和纯化多肽抗生素。在 2.5% TSB 液体培养基中培养 PLS 76，并在 70°C、150 转/分的条件下培养 112 小时，从而生产出抗生素。对离心后的粗上清液进行茚三酮检测，结果表明 PLS 76 产生了一种多肽抗生素。上清液用甲醇分馏。用 Kirby-Bauer 法检测抗生素活性，结果表明 PLS 76 对大肠杆菌和金黄色葡萄球菌的抑菌区约为 12 毫米。甲醇馏分经制备型薄层色谱法进一步纯化，产生了多个馏分。将具有相同 Rf 的馏分集中起来，进行茚三酮检测。经茚三酮检测呈阳性结果的馏分 A 对大肠杆菌和金黄色葡萄球菌的抑制区仅为 7 毫米。四极杆飞行时间质谱（QTOF）数据表明，该抗生素的结构与多肽类抗生素多粘菌素 B2 相似。

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Jurnal Riset Kimia

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12 weeks