Article

Characterisation of thermostable trypsin and determination of trypsin isozymes from intestine of Nile tilapia (Oreochromis niloticus L.)

Food Chemistry (Impact Factor: 3.39). 10/2012; 134(3):1533–1541. DOI: 10.1016/j.foodchem.2012.03.074

ABSTRACT

Trypsin from intestinal extracts of Nile tilapia (Oreochromis niloticus L.) was characterised. Three-step purification – by ammonium sulphate precipitation, Sephadex G-100, and Q Sepharose – was applied to isolate trypsin, and resulted in 3.77% recovery with a 5.34-fold increase in specific activity. At least 6 isoforms of trypsin were found in different ages. Only one major trypsin isozyme was isolated with high purity, as assessed by SDS-PAGE and native-PAGE zymogram, appearing as a single band of approximately 22.39 kDa protein. The purified trypsin was stable, with activity over a wide pH range of 6.0– 11.0 and an optimal temperature of approximately 55–60 °C. The relative activity of the purified enzyme was dramatically increased in the presence of commercially used detergents, alkylbenzene sulphonate or alcohol ethoxylate, at 1% (v/v). The observed Michaelis–Menten constant (Km) and catalytic constant (Kcat) of the purified trypsin for BAPNA were 0.16 mM and 23.8 s–1, respectively. The catalytic efficiency (Kcat / Km) was 238 s–1 mM–1.

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Available from: Krisna Rungruangsak-Torrissen
    • "Trypsin and trypsin-like enzymes demonstrate good potential for application in laundry detergents (Ktari et al., 2012), liquefaction of fish sauce (Klomklao, Benjakul, Visessanguan, Kishimura, & Simpson, 2006), recovering of carotenoids from shrimp wastes (Sila, Nasri, Bougatef, & Nasri, 2012) and production of protein hydrolysates with antioxidant activities and/or angiotensin-I converting enzyme (ACE) inhibitory activity (Khantaphant, Benjakula, & Kishimura, 2011; Nasri et al., 2013; Phanturat, Benjakul, Visessanguan, & Roytrakul, 2010). Different authors have reported the extraction and characterisation of alkaline proteases, especially trypsin and trypsin-like enzymes, from viscera of different species, such as zebra blenny (Ktari et al., 2012, Nile tilapia (Unajak et al., 2012), grey triggerfish (Jellouli et al., 2009), goby (Nasri et al., 2012), Giant Amazonian fish (Freitas-Junior et al., 2012) and farmed Mekong Giant catfish (Ketnawa, Benjakul, Ling, Martinez-Alvarez, & Rawdkuen, 2013; Rawdkuen, Vannabun, & Benjakul, 2012). Antioxidants are required to prevent lipid oxidation in food, avoiding the formation of toxic compounds and undesirable odours and flavours. "
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    No preview · Article · Feb 2016 · Food Chemistry
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    Full-text · Article · Aug 2015
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    • "In this study, no gonad was found in 400 g fish represented by the LJ stage, therefore, changes found in protease activity under different conditions could not be attributed to the onset of sexual maturity. In addition, some authors report enzyme changes during the ontogenesis of fish, suggesting that specific types of protease could be produced at a specific fish age by means of fish ontogenesis (Torrissen, 1987; Kuz᾽mina, 1996; Bassompierre et al., 1998; Chiu & Pan, 2002; Rathore et al., 2005; Chakrabarti et al., 2006; Unajak et al., 2012). "
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