Karakterisasi Plantarisin IIA-1A5 sebagai Antimikroba dan Evaluasi Aktivitas Sediaan Kering Beku Terenkapsulasi

Abstract

Bakteriosin adalah peptida dengan aktivitas antibakteri yang diproduksi oleh bakteri asam laktat dan digunakan sebagai pengawet alami. Penelitian sebelumnya menunjukkan bahwa Lactobacillus plantarum IIA-1A5 memproduksi bakteriosin yang diberi nama Plantarisin IIA-1A5 pada medium pertumbuhan yang dibuat dari whey yang diperkaya skim. Untuk aplikasi sebagai pengawet alami dan untuk memperbaiki masa simpan dan aktivitas anti mikrobanya, plantarisin perlu dienkapsulasi dan dikeringbekukan. Tujuan dari penelitian ini adalah untuk mengkarakterisasi dan mengevaluasi aktivitas antimikroba dari sediaan plantarisin IIA-1A5 yang terpurifikasi parsial dan terenkapsulasi kering beku. Ekstraksi dan purifikasi dari bakteriosin dimulai dengan presipitasi dengan ammonium sulfat, yang diikuti dengan dialysis, dan penukar kation kromatografi. Purifikasi parsial dari plantarisin kemudian dimikroenkapsulasi dengan maltodextrin kemudian dilanjutkan dengan proses kering beku. Berdasarkan pada SDS-PAGE, fraksi protein ke-7 (F7) dari plantarisin yang dipurifikasi parsial memiliki pita tunggal dan berat molekul sekitar 9,65 kDa. Konfirmasi lebih lanjut dengan menggunakan MALDI-TOF MS, ternyata pita tunggal tersebut terdiri dari 5 peptida yang diidentifikasi berbobot molekul masing-masing sebagai berikut 5,5, 7,80, 7,96, 9,09, dan 9,27 kDa. Plantarisin kering beku memiliki aktivitas antimikroba terhadap Staphylococcus aureus tiga kali lipat dibandingkan dengan aktivitas antimikroba dari supernatan bebas sel, dan lebih tinggi dibandingkan dengan nisin, namun kurang bila dibandingkan dengan antibiotik ampisilin dan penisilin. Kesimpulannya, aktivitas antimikroba plantarisin kering beku dapat ditentukan dan lebih tinggi dibandingkan dengan nisin, ampisilin dan penisilin.Characterization of Plantarisin IIA-1A5 as Antimicrobial subtances and Evaluation of Acitivity of Freeze-dried Microencapsulated PreparationAbstractBacteriocins are peptides with antibacterial activity produced by lactic acid bacteria and used as natural preservatives. Previous studies showed that Lactobacillus plantarum IIA-1A5 produces bacteriocin named plantaricin IIA-1A5 in the medium consisting whey enriched with skim milk. For application as food preservatives and to improve its shelf-lie and activity, plantaricin was needed to be microencapsulated and freeze dried. The objective of this research was to characterize and evaluate the activity of partially purified freeze dried microencapsulated plantaricin IIA-1A5. Characterisation of partially purified plantaricin IIA-IA5 includes the identification of active fractions and molecular weight, evaluation of activity at different stage of purification and evaluation of antimicrobial activity of freeze dried microencapsulated plantaricin IIA-IA5. Extraction and prificafication of the bacteriocins started with precipitacion with ammonium sulfate, followed by dialysis, and cation exchange chromatography. The partial purified of plantaricin was then microencapsulated in maltodextrin followed by freeze drying. Based on SDS-PAGE, the protein fraction F7 of partially purified plantaricin had a single band and molecular weight about 9.65 kDa. Further analyses using MALDI-TOF, it revealed that five peptides were identified from one single band plantaricin with molecular weight 5.5, 7.80, 7.96, 9.09, and 9.27 kDa, respectively. The freeze dried plantaricin freeze showed antimicrobial activity against Staphylococcus aureus three times stringer as compared to the activity of cell free supernatant, and was higher than nicin, but less than antibiotic ampicilin and penicilin. As concusion, the activity of freeze dried plantaricin could be determined and had a higher value than nicin, ampicilin and penicilin.

Application of Plantaricin as an Antimicrobial Substrate in the Milking Process to Maintain Milk Quality in Smallholder Dairy Farm

“…Gram positive bacteria were more sensitive, while gram negative bacteria were more resistant (Fatmarani et al 2018). Based on the research results of Soenarno et al (2020), that plantaricin IIA-1A5 has a broad spectrum antimicrobial ability against Gram positive and Gram negative pathogenic bacteria.…”

“…Arief et al (2015) reported that the molecular weight of crude plantaricin IIA-1A5 was less than 10 kDa and was classified in the IIA classification. The plantaricin IIA-1A5 was successfully purified using cation exchange chromatography, and had a molecular weight of 9.65 kDa (Soenarno et al 2020). This is similar to the study of Arifin et al (2020), that plantaricin IIA-1A5 was successfully purified from ammonium sulfate, and cation exchange chromatography and had a molecular weight of 9.4 kDa.…”

“…Application of 0.3% plantaricin could maintain the quality of beef meatballs for 20 hours at room temperature storage (Kia et al 2015) and effective as a preservative in beef sausage products for six days inside the cold storage (Arief et al 2017). Soenarno et al (2020) stated that plantarisin IIA-1A5 and nisin to fresh milk reduced the increase in the amount of TPC in the first two hours compared to controls. In terms of safety, these results indicated that iodine and plantaricin IIA-1A5 can act as antimicrobials to slow down the total microbial population in fresh milk during the milking process.…”

“…These bacteriocins are non-toxic to humans, stable to changes in pH and temperature, and safe for food preservatives because they are easily digested by digestive enzymes (Hata et al 2010). Therefore, bacteriocins can be used as a biopreservative in fresh and processed food products (Soenarno et al 2020). Lactobacillus plantarum is a bacteriocin-producing lactic acid bacterium known as plantaricin.…”