Optimization and characterization of enterocin Enterococcus faecalis K2B1 isolated from Toraja’s Belang Buffalo Milk, South Sulawesi, Indonesia

Alang, Hasria; Kusnadi, Joni; Ardyati, Tri; Suharjono, Suharjono

doi:10.13057/biodiv/d210351

Cited by 6 publications

(4 citation statements)

References 21 publications

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“…Thus the bacteriocins produced are proteins, polypeptides, and peptides with molecular weights above 10 kDa which are estimated to contain plantaricin (pln) peptides. A decrease in protein concentration after the purification stage was also reported by Alang et al (2020) for enterocin from Enterococcus faecalis K2B1 and Mulyawati et al (2019b) for plantaricin from L. plantarum SB7.…”

Section: Bacteriocin Protein Charactersupporting

confidence: 68%

“…Therefore, it was suspected that the precipitation results should also have a protein band profile detected. CFS protein band profiles and undetected precipi-tation were also experienced by the study and the results of dialysis detected a protein band profile (Mulyawati et al, 2019b;Alang et al, 2020). Protein bands formed from the results of the dialysis step can be detected through SDS-PAGE analysis.…”

Section: Protein Molecular Weightmentioning

confidence: 66%

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Detection of Plantaricin-Encoding Gene and Its Partial Purification in Lactobacillus plantarum BP102

Suryani,

Jatmiko,

Mustafa

2023

Jurnal Biodjati

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Lactobacillus plantarum BP102 isolated from garlic bulb tissue has probiotic properties, including producing bacteriocin called plantaricin. This study aimed to detect the gene encoding bacteriocin produced by Lactobacillus plantarum BP102, and to evaluate the bacteriocin activity at each stage of partial purification. After the end of the log phase of L. plantarum BP102 was determined, and the bacteriocin-encoding genes were checked by PCR technique. Partial purification of bacteriocin was elucidated including pH-neutralized cell-free-supernatant (CFS), precipitation using 80% of ammonium sulfate, and dialysis (cut-off 10 kDa), then the bacteriocin activity in every partial purification stage was evaluated. The molecular weight of plantaricin was estimated using SDS-PAGE analysis. Lactobacillus plantarum BP102 harbored the gene encoding plantaricin (pln) biosynthesis, namely plnEF and plnK genes. The activity of crude bacteriocin was inactivated by the presence of proteinase-K enzyme. The protein concentration was gradually decreased along with the purification process. The bacteriocin activity was demonstrated at each step of the purification process (CFS, precipitation, and dialysis) against Bacillus cereus by 9.23 ± 0.20 mm, 7.86 ± 0.15 mm, and 7.6 ± 0.10 mm, respectively; while, Escherichia coli by 10.3 ± 0.55 mm, 7.4 ± 0.1 mm, and 6.86 ± 0.45, respectively. The molecular weight of partially purified bacteriocin BP102 was found to be approximately 15.9 kDa. The overlaid part of the gel showed a slight inhibition against E. coli due to a low protein concentration. This bacteriocin purification process should be further optimized to improve the bacteriocin activity that could be useful for food preservation.

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Section: Bacteriocin Protein Charactersupporting

confidence: 68%

Section: Protein Molecular Weightmentioning

confidence: 66%

Detection of Plantaricin-Encoding Gene and Its Partial Purification in Lactobacillus plantarum BP102

Suryani,

Jatmiko,

Mustafa

2023

Jurnal Biodjati

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“…Enterocin Enterococcus faecalis KT11 mampu menghambat pertumbuhan bakteri Gram positif dan Gram negative seperti Klebsiella pneumoniae, Bacillus subtilis NRRL-NRS744, Pseudomonas aeruginosa, Staphylococcus aureus dan Staphylococcus epidermidis [22]. Enterocins Enterococcus durans MF5 dapat menghambat pertumbuhan Shiga-like toxin-producing Escherichia coli (STEC) [23] dan enterocin K2B1 yang dihasilkan oleh E. faecalis K2B1 dapat menghambat pertumbuhan S. aureus, B. cereus dan P. aeruginosa [8]. Beberapa jenis enterocin serta pemanfaatannya dalam bidang industri dapat terlihat pada Tabel 2.…”

Section: Enterococcusunclassified

Review : Enterocyn from Enterococcus Genus as a Probiotic, Antimicrobial and Biopreservative

Alang¹

2020

Pharmauho

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Lactic acid bacteria (LAB) is agras, produces antimicrobial compounds called bacteriocin. It can reduce its dependence on antibiotic use. Enterococcus is one of member LA.B. This bacterium has a short and paired chain shape and produces bacteriocin called enterocin. This compound can inhibit the growth of pathogenic microbes that cause disease and food spoilage, so that it can be used as an antimicrobial and biopreservative and also as a probiotic candidate. The use of Enterococcus as a probiotic and biopreservative is often disputed. However, some studies suggest that cases of bacteremia and urinary tract infections by the genus only occur in people who have immune compromised and enterococcus that have been resistant to vancomycin.

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“…Enterococcus faecalis K2B1 is an isolate obtained from Indonesian Toraja's Belang Buffalo milk and has been known to have potential as a probiotic. In addition, the optimum conditions for producing and Enterocins characteristics produced by these isolates have also been known to be potential as bio preservatives (Alang et al, 2020). However, studies related antimicrobial activity against S.typhi by E. faecalis and genes encoding enterocins from these isolates have not been reported.…”

Section: Introductionmentioning

confidence: 99%

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

Alang

Kusnadi

Ardyati

et al. 2020

BES

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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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Optimization and characterization of enterocin Enterococcus faecalis K2B1 isolated from Toraja’s Belang Buffalo Milk, South Sulawesi, Indonesia

Cited by 6 publications

References 21 publications

Detection of Plantaricin-Encoding Gene and Its Partial Purification in Lactobacillus plantarum BP102

Detection of Plantaricin-Encoding Gene and Its Partial Purification in Lactobacillus plantarum BP102

Review : Enterocyn from Enterococcus Genus as a Probiotic, Antimicrobial and Biopreservative

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

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