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Isolation of Lactic Acid Bacteria That Produce Protease and Bacteriocin-Like Substance From Mud Crab (Scylla sp.) Digestive Tract (Isolasi Bakteri Asam Laktat yang Menghasilkan Protease dan Senyawa Bacteriocin-Like dari Saluran Pencernaan Kepiting

1Department of Marine, Faculty of Fisheries and Marine, Kampus C Universitas Airlangga, , Indonesia

2Jl. Mulyorejo, Surabaya. 60115 Indonesia, Indonesia

Published: 1 Mar 2015.

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Abstract

Saluran pencernaan merupakan lingkungan kompleks yang terdiri atas berbagai spesies bakteri. Saluran pencernaan biota perairan terdiri atas bakteri aerob dan anaerob yang mampu memproduksi senyawa antibakteri dan enzim. Tujuan penelitian ini adalah untuk mengisolasi bakteri asam laktat yang menghasilkan protease dan senyawa bakteriosin-like dari saluran pencernaan kepiting bakau. Isolasi dan karakterisasi isolat dilakukan menggunakan media MRS. Supernatan netral bebas sel isolat telah diuji dengan menggunakan disc difusi agar terhadap bakteri patogen dan pembusuk. Uji produksi enzim protease telah diuji dengan metode disc diffusion agar menggunakan media kasein agar. Di antara seratus isolat, terdapat 96 isolat menunjukkan zona bening di MRS + CaCO3,, katalase negatif, dan bakteri Gram positif. Tiga puluh empat isolat bakteri asam laktat menghasilkkan protease dan hanya empat isolat (yaitu IKP29, IKP30, IKP52, dan IKP94) menunjukkan penghambatan yang kuat terhadap bakteri patogen dan pembusuk. Terdapat tiga pola inhibisi dari keempat isolate terhadap Bacillus subtilis, Staphylococcus aureus, Eschericia coli, dan Salmonella sp. Empat isolat tersebut berpotensi untuk dimanfaatkan sebagai starter pada produksi fermentasi produk hasil perikanan. Penelitian ini adalah penelitian pertama terkait isolasi bakteri asam laktat yang menghasilkan protease dan bakteriosin dari saluran pencernaan dari kepiting bakau.

Kata kunci: Bakteri Asam Laktat, Bakteriosin-like substance, Protease, Scylla  sp.

Digestive tract is complex environment consist of large amount of bacteria’s species. Fish intestine bacteria consist of aerobic or facultative anaerob bacteria which can produce antibacterial and enzym. The objectives of this research were to isolated lactic acid bacteria that produce bacteriocin-like and protease from mud crab digestive tract. Isolation and characterization of isolates were conducted employing media MRS.  Neutralized cell free supernatant of isolates were tested using disc diffusion agar of against pathogenic and spoilage bacteria to indicate bacteriocin-like-producing lactic acid bacteria. Protease-producing isolate was tested using disc diffusion method in casein agar. Among a hundred isolates, 96 isolates were showed clear zone in MRS+CaCO3,, catalase negative, and Gram positive bacteria. Thirty four isolates produced protease and only four isolates (i.e. IKP29, IKP30, IKP52, and IKP94) showed strong inhibition against pathogenic and spoilage bacteria. There were three patterns of inhibition among three isolates against Bacillus subtilis, Staphylococcus aureus, Eschericia coli, and Salmonella sp. All three isolates showed potential uses for produce starter culture for fishery product fermentation purpose. This is the first report of isolation lactic acid bacteria that produced protease and bacteriocin-like from digestive tract of mud crab.

Keywords: Lactic acid bacteria, Bacteriocin-like substance, Protease, Scylla  sp.
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