Sains Malaysiana 43(11)(2014): 1715–1723

Optimization of Enzymatic Hydrolysis of Tilapia Muscle (Oreochromis niloticus) using Response Surface Methodology (RSM)

(Pengoptimuman Hidrolisis Berenzim bagi Otot Tilapia (Oreochromis niloticus) Menggunakan Kaedah Sambutan Permukaan)

JUMARDI ROSLAN¹, SITI MAZLINA MUSTAPA KAMAL¹*, KHAIRUL FAEZAH MD. YUNOS¹ & NORHAFIZAH ABDULLAH²

¹Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia

43400 Serdang, Selangor, Malaysia

²Department of Chemical and Environmental Engineering, Faculty of Engineering,

Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia

Diserahkan: 2 September 2013/Diterima: 14 April 2014

ABSTRACT

Fish protein hydrolysate was prepared from tilapia muscle using commercial Alcalase enzyme. Optimization of enzymatic hydrolysis process for preparing tilapia muscle protein hydrolysates (TMPH) was performed by employing central composite design (CCD) method of response surface methodology (RSM). O-phtaldialdehyde (OPA) method was employed to calculate the degree of hydrolysis (DH), which is the key parameter for monitoring the reaction of protein hydrolysis. The suggested model equation was proposed based on the effects of pH, temperature, substrate concentration and enzyme concentration on the DH. Optimum enzymatic hydrolysis conditions using Alcalase enzyme were obtained at pH7.5, temperature of 50^oC, substrate concentration of 2.5% and enzyme concentration of 4.0%. Under these conditions, the highest value of the DH was achieved at 25.16% after hydrolysing at 120 min. The TMPH was further assessed for their nutritional value with respect to chemical and amino acid compositions. Molecular weight distributions of TMPH were characterized by SDS-PAGE. TMPH contains moderate amount of protein (28.14%) and good nutritive value with respect to the higher total amino acid composition (267.57 mg/g). Glutamic acid, aspartic acid and lysine were the most abundant amino acids present in TMPH with values 42.68, 29.16 and 26.21 mg/g, respectively. Protein hydrolysates from tilapia muscle containing a desirable peptide with low molecular weight which may potentially to be used as functional food products.

Keywords: Alcalase; degree of hydrolysis; enzymatic hydrolysis; response surface methodology; tilapia muscle

ABSTRAK

Protein ikan hidrolisat disediakan daripada otot tilapia dengan menggunakan enzim alkalase. Pengoptimuman proses hidrolisis berenzim untuk penyediaan protein hidrolisat otot tilapia (TMPH) dilakukan dengan menggunakan reka bentuk komposit berpusat (CCD) iaitu kaedah sambutan permukaan (RSM). Kaedah O-phtaldialdehid (OPA) telah digunakan untuk penentuan darjah hidrolisis (DH) yang merupakan kunci utama untuk memantau tindak balas hidrolisis protein. Model cadangan yang digunakan adalah berasaskan kepada kesan pH, suhu, kepekatan substrat dan kepekatan enzim ke atas DH. Keadaan hidrolisis optimum bagi enzim alcalase diperoleh pada pH7.5, suhu 50^oC, kepekatan substrat 2.5% dan kepekatan enzim 4.0%. Pada keadaan ini, nilai tertinggi DH dicapai pada 25.16% selepas hidrolisis dijalankan selama 120 min. TMPH seterusnya diuji nilai nutrisinya berdasarkan kepada komposisi kimia dan asid amino. Taburan berat molekul TMPH dicirikan menggunakan SDS-PAGE. TMPH mengandungi kandungan protein yang sederhana (28.14%) serta nilai nutrisi yang baik berdasarkan kepada jumlah asid amino yang tinggi (267.57 mg/g). Asid glutamik, asid aspartik dan lisina merupakan asid amino yang paling banyak terkandung di dalam TMPH dengan nilai masing-masing adalah 42.68, 29.16 dan 26.21 mg/g. Protein hidrolisat daripada daging tilapia mengandungi peptida yang dikehendaki dengan berat molekul rendah yang berpotensi untuk digunakan sebagai produk makanan berfungsi.

Kata kunci: Alkalase; darjah kehidrolisisan; hidrolisis berenzim; kaedah sambutan permukaan; otot tilapia

RUJUKAN

Abdul-Hamid, A., Bakar, J. & Bee, G.H. 2002. Nutritional quality of spray dried protein hydrolysate from Black Tilapia (Oreochromis mossambicus). Food Chemistry 78: 69-74.

Adler-Nissen, J. 1986. Enzymatic Hydrolysis of Food Proteins. Oxford, UK: Elsevier Applied Science Publishers.

Adler-Nissen, J. 1979. Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzenesulfonic acid. Journal of Agricultural and Food Chemistry 27(6): 1256-1262.

Amiza, M.A., Nurul Ashikin, S. & Faazaz, A.L. 2011. Optimization of enzymatic protein hydrolysis from silver catfish (Pangasius sp.) frame. International Food Research Journal 18: 775-781.

Association of Official Analytical Chemists (AOAC). 2005. Official Methods of Analysis. 16th ed. Washington DC, USA: AOAC Inc.

Aspmo, S.I., Horn, S.J. & Eijsink, V.G.H. 2005. Enzymatic hydrolysis of Atlantic cod (Gadus morhua L.) viscera. Process Biochemistry 40: 1957-1966.

Benjakul, S. & Morrissey, M.T. 1997. Protein hydrolysates from pacific whiting solid wastes. J. Agric. Food Chem. 45: 3423-3430.

Bhaskar, N., Benila, T., Radha, C. & Lalitha, R.G. 2008. Optimization of enzymatic hydrolysis of visceral waste proteins of Catla (Catla catla) for preparing protein hydrolysate using a commercial protease. Bioresource Technology 99: 335-343.

Church, F.C., Swaisgood, H.E., Porter, D.H. & Catignani, G.L. 1983. Spectrophotometric assay using o-phthaldialdehyde for determination of proteolysis in milk and isolated milk proteins. Journal of Dairy Science 66: 1219-1227.

Foh, M.B.K., Qixing, J., Amadou, I. & Xia, W.S. 2010. Influence of ultrafiltration on antioxidant activity of tilapia (Oreochromis niloticus) protain hydrolysate. Advanced Journal of Food Science and Technology 2(5): 227-235.

Joglekar, A.M. & May, A.T. 1987. Product excellence through design of experiments. Cereal Foods World 32(12): 857-868.

Kristinsson, H.G. & Rasco, B.A. 2000. Biochemical and functional properties of Atlantic Salmon (Salmo salar) muscle proteins hydrolyzed with various alkaline proteases. Journal of Agricultural and Food Chemistry 48: 657-666.

Lalasidis, G., Bostrom, S. & Sjoberg, L.B. 1978. Low molecular weight enzymatic fish protein hydrolysates: Chemical composition and nutritive value. Journal of Agricultural and Food Chemistry 26(3): 751-756.

Liaset, B., Lied, E. & Espe, M. 2000. Enzymatic hydrolysis of by-products from the fish-filleting industry; Chemical characterisation and nutritional evaluation. Journal of the Science of Food and Agriculture 80: 581-589.

Nielsen, P.M., Petersen, D. & Dambmann, C. 2001. Improved method for determining food protein degree of hydrolysis. Journal of Food Science 66(5): 642-646.

Normah, I., Jamilah, B., Saari, N. & Che Man, Y. 2005. Optimization of hydrolysis conditions for the production of threadfin bream (Nemipterus japonicus) hydrolysate by Alcalase®. Journal of Muscle Foods 16: 87-102.

Pacheco-Aguilar, R., Mazorra-Manzano, M.A. & Ramirez- Suarez, J.C. 2008. Functional properties of fish protein hydrolysates from Pacific whiting (Merluccius productus) muscle produced by a commercial protease. Food Chemistry 109: 782-789.

Schagger, H. & von Jagow, G. 1987. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Analytical Biochemistry 166: 368-379.

See, S.F., Hoo, L.L. & Babji, A.S. 2011. Optimization of enzymatic hydrolysis of Salmon (Salmo salar) skin by Alcalase. International Food Research Journal 18(4): 1359-1365.

Shahidi, F., Han, X.Q. & Synowiecki, J. 1995. Production and characteristics of protein hydrolysates from capelin (Mallotus villosus). Food Chemistry 53: 285-293.

Shamloo, M., Bakar, J., Mat Hashim, D. & Khatib, A. 2012. Biochemical properties of red Tilapia (Oreochromis niloticus) protein hydrolysates. International Food Research Journal 19(1): 183-188.

Thiansilakul, Y., Benjakul, S. & Shahidi, F. 2007. Compositions, functional properties and antioxidative activity of protein hydrolysates prepared from round scad (Decapterusmaruadsi). Food Chemistry 103: 1385-1394.

Venugopal, V., Chawla, S.P. & Nair, P.M. 1996. Spray dried protein powder from threadfin beam: Preparation, properties and comparison with FPC type-B. J. Muscle Foods 7: 55-71.

Wangtueai, S. & Noomhorm, A. 2009. Processing optimization and characterization of gelatin from lizardfish (Saurida spp.) scales. LWT - Food Science and Technology 42: 825-834.

*Pengarang untuk surat-menyurat; email: smazlina@upm.edu.my