Purifikasi Parsial dan Karakterisasi B-Galactosidase dari Lactobacillus plantarum Strain D-210

Nur Nunu Prihantini, Tatik Khusniati, Maria Bintang, Abdul Choliq, Sulistiani Sulistiani

Abstract


Pemurnian parsial dan karakterisasi B-galaktosidase dari Lactobacillus plantarum strain D-210 belum dilaporkan. L. plantarum strain D-210 ditemukan sebagai bakteri penghasil B-galaktosidase sebagian dimurnikan dengan dialisis. Hasil penelitian menunjukkan bahwa aktivitas optimum dalam 24 jam dengan dan total protein adalah 0,454 mg/ml pada pH 6.5 aktivitas enzim 252,341 U/ml, dan suhu 45°C dengan aktivitas 0,582 U/ ml. Total aktivitas B-galaktosidase L.plantarum strain D-210 adalah 138,396 U dan endapan dengan amonium sulfat dicapai pada 40% - 50% dengan aktivitas total 87,030 U. Setelah dialisis, aktivitas total adalah 50,420 U. Penghambat B-galaktosidase adalah Hg dan Cu dengan aktivitas relatif adalah 56,82% dan 1,04%, sedangkan aktivator adalah Mg, Mn, Ca, Co, Zn. Vmaks dari enzim adalah 0.093 µmol/menit dan KM enzim B-galaktosidase L. plantarum adalah 0,491 mM. Berdasarkan karakteristik B-galaktosidase, dapat disimpulkan bahwa L. plantarum strain D-210 adalah bakteri baik dan unggul yang dapat memproduksi B-galaktosidase. Studi lebih lanjut diperlukan untuk mengeksplorasi kemungkinan menggunakan bakteri ini dalam pengolahan susu pada bayi dengan intoleransi laktosa.


Partial purification and characterization of Lactobacillus plantarum strain D-210 B-galactosidase has not been reported yet. L. plantarum strain D-210 known as bacteria producing B-galactosidase was partially purified by membrane dialysis. The results showed that optimum activity in 24 hour with total protein yield 0.454  mg/ml at pH 6.5  the enzyme activity was 252.341 U/ml, and at 45°C the activity was 0.582 U/ml. The total activity of B-galactosidase L.plantarum strain D-210 was 138.396 U and precipitated by sulphate ammonium at 40%-50% with total activity was 87.030 U. Following dialysis, the total activity was 50.420 U. The inhibitors of B-galactosidase were Hg and Cu with relative enzyme activities of 56.82% and 1.04% respectively, while the activators were Mg,Mn,Ca,Co,Zn. Vmax of the enzyme was 0.093 µmol/min and KM was 0.491 mM. Based on the characteristics of the enzyme, it can be concluded that L. plantarum strain D-210 was a good B-galactosidase producing bacteria. Further studies are required to explore the possibility of using this bacteria in milk processing for lactose intolerance babies.


Keywords


B-galactosidase; partial purification; total activity; galactosidase inhibitor

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DOI: https://doi.org/10.33476/jky.v21i1.16

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