Isolation and Characterization of Plantaricin Produced by Lactobacillus plantarum Strains (IIA-1A5, IIA-1B1, IIA-2B2)

Abstract

Bacteriocins produced by Indonesian lactic acid bacteria Lactobacillus plantarum IIA-1A5, IIA-1B1, IIA-2B2 were purified and characterized. Plantaricin W gene had been successfully amplified from all strains. This amplicon showed the expected 200 bp size of plantaricin W gene. This bacteriocins purified from L. plantarum IIA-1A5, IIA-1B1, and IIA-2B2 were named plantaricin IIA-1A5, IIA-1B1, and IIA-2B2. Purification by cation exchange chromatography increased the purity (fold) and activity of plantaricins. Purity of plantaricin IIA-1A5 was increased by 3.13 fold with specific activity 13.40 AU/mg. Plantaricin IIA-1B1 had 2.98 fold purity with specific activity 5.12 AU/mg, while purity of plantaricin IIA-2B2 was 1.37 fold with specific activity 7.70 AU/mg. All plantaricins could inhibit the growth of pathogenic bacteria, such as Escherichia coli, Salmonella typhimurium, Bacillus cereus, and Staphylococcus aureus. Plantaricins could be digested by trypsin. Stability of plantaricins at 80 oC for 30 min and at 121 oC for 15 min were affected by type of plantaricin and species of pathogenic bacteria. Generally, plantaricin IIA-1A5 was better as antimicrobial agent than plantaricin IIA-1B1 and plantaricin IIA-2B2

Bakteriosin dihasilkan oleh Lactobacillus plantarum IIA-1A5, IIA-1B1, IIA-2B2, bakteri asam laktat (BAL) asal Indonesia, yang dipurifikasi dan dikarakterisasi. Gen plantarisin W berhasil diam-plifikasi dari semua strain dengan ukuran basa 200 pb. Bakteriosin yang dipurifikasi dari L. planta-rum IIA-1A5, IIA-1B1, dan IIA-2B2 dinamakan plantarisin IIA-1A5, IIA-1B1, dan IIA-2B2. Purifikasi dengan menggunakan kromatografi pertukaran kation mampu meningkatkan aktivitas plantarisin tersebut. Kemurnian plantarisin IIA-1A5 meningkat sebesar 312.65% dengan aktivitas spesifik 13.40 AU/mg. Plantarisin IIA-1B1 mempunyai tingkat kemurnian sebesar 297.72% dengan aktivitas spesifik 5.12 AU/mg, sedangkan peningkatan kemurnian produk plantarisin IIA-2B2 sebesar 137.48% dengan aktivitas spesifik 7.70 AU/mg. Karakterisasi plantarisin IIA-1A5, IIA-1B1 dan IIA-2B2 menunjukkan bahwa plantarisin tersebut mampu menghambat pertumbuhan bakteri patogen Escherichia coli, Salmonella typhimurium, Bacillus cereus, dan Staphylococcus aureus dan mampu didegradasi oleh tripsin. Stabilitas terhadap perlakuan pemanasan pada suhu pasteurisasi (80 oC selama 30 menit) dan sterilisasi (121 oC selama 15 menit) dipengaruhi oleh jenis plantarisin serta jenis bakteri patogen yang dihambat pertumbuhannya. Berdasarkan karakteristiknya secara umum, plantaricin IIA-1A5 mem-punyai sifat antimikroba yang lebih baik dibandingkan dengan plantaricin IIA-1B1 dan plantaricin IIA-2B2.