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<p>The growing threat of antimicrobial resistance in oral pathogens necessitates safe and sustainable alternatives to conventional therapies. Enterocin CC2, a novel bacteriocin from <i>Enterococcus faecium</i> CC2, was purified using an optimized polyethylene glycol (PEG)–citrate aqueous two-phase system (ATPS). The process achieved a purification factor of 4.73 and recovery yield of 73.33%, which improved to 5.13 and 84.61% with the addition of 4% NaCl. SDS-PAGE revealed a molecular size of ~ 5.5–6&#xa0;kDa, and identity confirmation was obtained via HPLC and LC–MS. Enterocin CC2 displayed potent antimicrobial activity against <i>Streptococcus mutans</i> with MIC and MBC values of 0.50 and 1.00&#xa0;mg/mL, respectively, and demonstrated faster bactericidal action than 0.2% chlorhexidine. ISO 10993–5 cytotoxicity testing confirmed its safety on 3T3-L1 cells at concentrations ≤ 1&#xa0;mg/mL. These findings highlight enterocin CC2 as a promising candidate for oral health applications and establish PEG–citrate ATPS as a scalable, eco-friendly platform for bacteriocin purification.</p>

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Therapeutic Potential and Sustainable Bioprocessing of Enterocin CC2: A Probiotic-Derived Antimicrobial Targeting Streptococcus mutans in Oral Health

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年9月18日

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  • Zhang Jin Ng,
  • Chee-Yuen Gan,
  • Mazni Abu Zarin,
  • Chee Keong Lee,
  • John Chi-Wei Lan,
  • Hui Suan Ng,
  • Joo Shun Tan

摘要

The growing threat of antimicrobial resistance in oral pathogens necessitates safe and sustainable alternatives to conventional therapies. Enterocin CC2, a novel bacteriocin from Enterococcus faecium CC2, was purified using an optimized polyethylene glycol (PEG)–citrate aqueous two-phase system (ATPS). The process achieved a purification factor of 4.73 and recovery yield of 73.33%, which improved to 5.13 and 84.61% with the addition of 4% NaCl. SDS-PAGE revealed a molecular size of ~ 5.5–6 kDa, and identity confirmation was obtained via HPLC and LC–MS. Enterocin CC2 displayed potent antimicrobial activity against Streptococcus mutans with MIC and MBC values of 0.50 and 1.00 mg/mL, respectively, and demonstrated faster bactericidal action than 0.2% chlorhexidine. ISO 10993–5 cytotoxicity testing confirmed its safety on 3T3-L1 cells at concentrations ≤ 1 mg/mL. These findings highlight enterocin CC2 as a promising candidate for oral health applications and establish PEG–citrate ATPS as a scalable, eco-friendly platform for bacteriocin purification.