Antimicrobial Activity and Identification of Gene Encoding Enterocin Enterococcus faecalis K2B1 Isolated from Toraja's Belang Buffalo Milk

H. Alang, J. Kusnadi, T. Ardyati, Suharjono
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Abstract

Background: Enterocin in Enterococcus is coded by enterocin encoding genes namely A, B, P and L50A / B. The purpose of this study was to identify enterocin gene encoding enterococcus faecalis K2B1 probiotic candidate from Belang Toraja buffalo milk and antimicrobial activity to S. typhi. Methods: identification of enterocin gene encoding using ent A, B, P and L50A / B, partial purification using ammonium sulfate on 80 % concentration and antimicrobial activity against to Salmonella typhi using disk diffusion method. The results of PCR amplification are then sequenced and BLASTX on NCBI. Result: Antimicrobial activity of Precipitate and crude against S. typhi are 193 and 201 respectively. Identification gene encoding enterocin shows that Ent A, B and P cannot be amplified and only EntL50A / B can be amplified with a sequence size of 86 bp. The sequence of enterocin encoding genes in E. faecalis K2B1 has 94% similarity with hypothetical protein EB34_00789 E. faecalis on GenBank with accession number RBR60004.1 Conclusion: EntL50A / B E. faecalis K2B1 has a size of 86 bp and is 94% identical to the hypothetical protein EB34_00789 and Enterocin can be used as antimicrobial or bio preservative.    
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Toraja Belang水牛奶中粪肠球菌K2B1的抑菌活性及基因鉴定
背景:肠球菌中的肠毒素由编码肠毒素的基因A、B、P和L50A/B编码。本研究的目的是从Belang Toraja水牛奶中鉴定编码粪肠球菌K2B1益生菌候选物的肠毒素基因和对伤寒杆菌的抗菌活性。方法:用ent A、B、P和L50A/B对肠肽基因进行编码鉴定,用80%浓度的硫酸铵进行部分纯化,用纸片扩散法对伤寒沙门氏菌进行抗菌活性鉴定。然后对PCR扩增的结果进行测序,并在NCBI上进行BLASTX。结果:沉淀液和粗品对伤寒杆菌的抗菌活性分别为193和201。编码肠毒素的鉴定基因显示Ent A、B和P不能被扩增,只有EntL50A/B可以被扩增,序列大小为86bp。粪肠球菌K2B1中编码肠毒素的基因序列与GenBank上登录号为RBR60004.1的假定蛋白EB34_00789具有94%的相似性。
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8 weeks
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