Identifikasi Bakteri dan Purifikasi Partial Bakteriosin dari Bakteri Asam Laktat GMP 1 serta Karakterisasi Aktivitas Antibakterinya terhadap Bakteri Kontaminan Produk Hasil Perikanan

Muhammad Yaafi' Al-Hammam, Indun Dewi Puspitas, S.P., M.Sc., Ph.D.; Mgs. M. Prima Putra, S.Pi., M.Sc., Ph.D.

2022 | Tesis | MAGISTER ILMU PERIKANAN

Isolat GMP 1 merupakan bakteri asam laktat (BAL) yang berhasil diisolasi dari pakasam. Penelitian ini bertujuan untuk mengidentifikasi isolat GMP 1 dan mengisolasi serta mengkarakterisasi bakteriosinnya. Identifikasi molekuler dengan primer spesifik BAL L5 15F (5'-GCTCAGGAYGAACGCYGG3') dan 687R (3'-CACCGCTACACATGRADTTC-5') berhasil mengidentifikasi isolat GMP 1 sebagai Lactobacillus sp. dengan kemiripan homolog 99.71 persen terhadap strain Lactobacillus sp. SDCM 1015. Bakteriosin Lactobacillus sp. GMP 1 dipurifikasi secara parsial dengan garam ammonium sulfat dengan tingkat kejenuhan 40 hingga 80 persen, dan dilanjutkan dengan dialisis. Bakteriosin murni didapatkan pada tingkat kejenuhan 80 persen. Bakteriosin yang diproduksi Lactobacillus sp. GMP 1 memiliki rentang stabilitas yang luas terhadap suhu 40 derajat celcius hingga 121 derajat celcius, pH 2 hingga 7, dan mampu mempertahakan aktivitas penghambatannya setelah mengalami penambahan enzim lisozim. Bakteriosin ini juga mampu menghambat pertumbuhan bakteri Gram positif, Staphylococcus aureus ATCC 5638 dan bakteri Gram negatif, Salmonella sp. 230C yang merupakan bakteri kontaminan pangan dengan nilai minimum inhibitory concentration (MIC) masing-masing sebesar 250 mikrogram per mililiter.

GMP 1 isolate is a lactic acid bacteria (LAB) that was successfully isolated from pakasam. This study aims to identify GMP 1 isolates and isolate and characterize their bacteriocins. Molecular identification with specific primers BAL L5 15F (5'-GCTCAGGAYGAACGCYGG3') and 687R (3'-CACCGCTACACATG RADTTC-5") succeeded in identifying GMP 1 isolate as Lactobacillus sp. with 99.71 percent identify to Lactobacillus sp. SDCM 1015. Bacteriocin produced by Lactobacillus sp. GMP 1 was partially purified with ammonium sulfate salt with a saturation level of 40 until 80 percent and followed by dialysis. The pure bacteriocin was obtained at a saturation level of 80 percent and had an inhibitory activity of 4897.75 AU against Staphylococcus aureus ATCC 5638. The bacteriocin produced by Lactobacillus sp. GMP 1 has a wide stability range at 40 celcius degree until 121 celcius degree, pH 2 until 7, and is able to maintain its inhibitory activity after being subjected to the addition of the enzyme lysozyme. This bacteriocin is also able to inhibit the growth of Gram-Positive bacteria, Staphylococcus aureus ATCC 5638 and Gram-Negative bacteria, Salmonella sp. 230C which is a food contaminant bacteria with a minimum inhibitory concentration (MIC) value of 250 microgram per mililiter each.

Kata Kunci : bakteriosin, karakterisasi, Lactobacillus sp. GMP 1, purifikasi/ bacteriocin, characterization, Lactobacillus sp. GMP1, purification