ANTIBACTERIAL POTENCY OF CELL FREE SUPERNATANT PRODUCED BY LACTIC ACID BACTERIA ISOLATED FROM INDONESIAN FERMENTED FISH PRODUCTS AGAINST HISTAMINE-PRODUCING BACTERIA

Masagus Muhammad Prima Putra, Muhammad Yaafi Al-Hamam, Giffarri Ahsan, Klara Kharisma Bunga Chandra, I. D. Puspita
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Abstract

The toxicity caused by high histamine content produced by histamine-producing bacteria (HPB) during the fermentation of scombridae fish group based fermented fish products is still a problem that requires a solution. This study aims to explore the potential of secondary metabolites in the form of cell free supernatant (CFS) produced by lactic acid bacteria (LAB) as antibacterial agents against HPB. The LAB was isolated from the fermented fishery products named bekasan, cincalok and fish sauce using MRS-Agar and fermented on MRS-broth for 48 hours. CFS was collected by centrifugation at 15,000 x g for 15 minutes, followed by heating at 100oC for 3 minutes and pH neutralization with 0.1N NaOH. Antimicrobial activity of CFS then tested on HFB namely Morganella morganii TK7, Citrobacter freundii CK1, and Klebsiella sp. CK13.2 (collection of the Laboratory of Fisheries Product Quality and Safety, Department of Fisheries UGM) using the macrodilution method. Isolation of LAB from all three products successfully isolated 34 isolates. The results of the antibacterial activity showed that 4 isolates namely GMCN 1.12, GMBK 2.6, GMBK 2.7, dan GMKI 2.1 were able to inhibit HFB growth more than 70%. The highest activity was shown by GMBK 2.7 which inhibits 98% against Morganella morganii TK7, 99% against Citrobacter freundii CK1, and 84% against Klebsiella sp. CK13.2. The antimicrobial activity was reduced after proteolytic enzymes were added suggesting that the bioactive compound came from peptide-based substances like bacteriocin.
从印尼发酵鱼制品中分离的乳酸菌制备的无细胞上清液对产组胺菌的抑菌效果
产组胺菌(HPB)在发酵鲭鱼群发酵鱼制品过程中产生的高组胺含量对鱼的毒性是一个亟待解决的问题。本研究旨在探讨乳酸菌(LAB)产生的无细胞上清(CFS)形式的次级代谢物作为HPB抗菌药物的潜力。采用mrs -琼脂法从发酵的鱼产品bekasan、cincalok和鱼露中分离出乳酸菌,在MRS-broth上发酵48 h。收集CFS, 15000 x g离心15分钟,100℃加热3分钟,0.1N NaOH中和pH。然后用大量稀释法测试CFS对HFB的抑菌活性,即摩根氏摩根氏菌TK7、弗氏柠檬酸杆菌CK1和克雷伯氏菌CK13.2(收集自渔农处渔产品质量安全实验室)。从三种产品中分离出34株分离物。抑菌活性结果表明,GMCN 1.12、GMBK 2.6、GMBK 2.7和GMKI 2.1 4株菌株对HFB的抑制作用均在70%以上。gmbk2.7对Morganella morganii TK7的抑制率为98%,对Citrobacter freundii CK1的抑制率为99%,对Klebsiella sp. CK13.2的抑制率为84%。加入蛋白水解酶后,抗菌活性降低,表明其生物活性化合物来自于细菌素等肽类物质。
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