THE POTENCY OF CHITOSAN AS AN ELICITOR ON ANTIBACTERIAL ACTIVITY OF Streptomyces sp. GMR-22 AGAINST HISTAMINE-PRODUCING BACTERIA

Mohamad Aji Ikhrami, J. Widada, I. D. Puspita, Masagus Muhammad Prima Putra
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Abstract

Streptomyces is a Gram-positive bacteria that produces the largest secondary metabolite compounds. The results of whole-genome sequence analysis showed that Streptomyces can carry more than 30 Biosynthetic Gene Clusters (BGC) encoding secondary metabolites that have the potential to be explored in the exploration for new bioactive compounds. However, not all BGC can be expressed in the laboratory scale and requires a specific activation method. This study aims to explore the potential of chitosan as an elicitor compound to activate and or increase the antibacterial activity of Streptomyces sp. GMR-22 was tested against histamine-producing bacteria (HPB) Morganella morganii TK7 and Citrobacter freundii CK1. Chitosan was added to the fermentation medium with the final concentration of 250, 500, and 750 µg/ml while without the addition of chitosan used as control. Fermentation was carried out for 10 days at room temperature, with constant agitation 200 rpm. The supernatant was separated by centrifugation at 3500 rpm for 15 minutes, then fractionation with ethyl acetate, concentrated by vacuum rotary evaporator, and freeze-dried. The test for antibacterial activity was carried out by the microdilution method with an extract concentration of 100 mg/ml. The test results of the microdilution method showed that the addition of chitosan successfully increases the antibacterial activity with the highest activity shown by the water fraction of 250 µg/ml addition of chitosan which effective in inhibiting the growth of Morganella morganii TK7 and Citrobacter freundii CK1 by 97,29% and 97,92% respectively.
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壳聚糖对链霉菌GMR-22对组胺产生菌抑菌活性的影响
链霉菌是一种革兰氏阳性细菌,产生最大的次级代谢物化合物。全基因组序列分析结果表明,链霉菌可携带30多个编码次生代谢产物的生物合成基因簇(BGC),这些基因簇在探索新的生物活性化合物方面具有潜力。然而,并不是所有的BGC都能在实验室规模上表达,需要特定的激活方法。本研究旨在探索壳聚糖作为激发子化合物激活或增加链霉菌的抑菌活性的潜力。GMR-22对组胺产生菌Morganella morganii TK7和Citrobacter freundii CK1进行了抑菌试验。在发酵培养基中分别添加250、500和750µg/ml的壳聚糖,同时不添加壳聚糖作为对照。在室温下发酵10天,持续搅拌200 rpm。上清液3500 rpm离心分离15分钟,与乙酸乙酯分馏,真空旋转蒸发器浓缩,冷冻干燥。采用微量稀释法,提取液浓度为100 mg/ml进行抑菌活性试验。微量稀释法测试结果表明,壳聚糖的加入成功地提高了抗菌活性,其中250µg/ml的壳聚糖水分数的抑菌活性最高,对莫organella morganii TK7和freundii Citrobacter CK1的抑制作用分别为97、29%和97、92%。
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