Optimization of Enzymatic Extraction of ACE Inhibitory Peptide from Rohu (Labeo rohita) Fish Waste using RSM

DOI: 10.18805/IJAR.B-4542    | Article Id: B-4542 | Page : 673-679
Citation :- Optimization of Enzymatic Extraction of ACE Inhibitory Peptide from Rohu (Labeo rohita) Fish Waste using RSM.Indian Journal of Animal Research.2022.(56):673-679
Vikas Kumar, R. Jeya Shakila, A.U. Muzaddadi, G. Jeyasekaran, D. Sukumar, P. Padmavathy, Yogesh Kumar vikas.kumar5@icar.gov.in
Address : ICAR-Central Institute of Post-Harvest Engineering and Technology, Ludhiana-141 004, Punjab, India.
Submitted Date : 25-05-2021
Accepted Date : 27-07-2021


Background: Hypertension is one of the cardiovascular disease that kills people silently across the globe. It can be controlled, in one of the way, by ACE inhibitory peptide extracted from aquatic resources. 
Methods: Rohu (Labeo rohita) fish wastes were quantified for their anatomical yield; analyzed for their proximate composition and optimized the enzymatic extraction parameters for ACE inhibitory peptides. Response surface methodology with Box-Benhken Design (RSM-BBD) was used to optimize alcalase concentration (0.5-2% v/w), hydrolysis temperature (45-60°C), hydrolysis time (60-240 min.) and solid: liquid (S/L) ratio (0.2-1) to obtain rohu fish waste peptides. 
Results: More waste generated in smaller (49.4%) than medium and bigger (34.5%) fish. Quantum of edible flesh (59.06%) was followed by head (23.9%), trimmings (5.18%), scales (4.19%) and swim bladder (0.65%). However, protein content was highest in swim bladder (34.1%) followed by scales (22.9%), trimmings (18.7%) and head (17.1%). Alcalase concentration (1.08%, v/w), temperature (52.10°C), hydrolysis time (129.18 min) and S/L ratio (0.8:1) were found optimum for extraction ACE inhibitory peptide with DH, ACE inhibition and PY of 19.27%, 54.98% and 51.37% respectively. Results showed the potential of extracted ACE inhibitory peptide as ingredients in functional food.


ACE inhibitory peptide Alcalase Anatomical yield Enzymatic ecxtraction Optimization Proximate composition Rohu fish waste


  1. 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.
  2. Baharuddin, N.A., Halim, N.R.A. and Sarbon, N.M. (2016). Effect of degree of hydrolysis (DH) on the functional properties and angiotensin I-converting enzyme (ACE) inhibitory activity of eel (Monopterus sp.) protein hydrolysate. International Food Research Journal. 23(4).
  3. Coleman, J.J. and Cox, A.R. (2012). Antihypertensive drugs. In JK Aronson (Ed), Side Effects of Drugs, Annual. 34 (pp. 317-332) (Chapter 20).
  4. Gao, X., Li, X., Yan, P., Sun, R., Kan, G. and Zhou, Y. (2018). Identification and functional mechanism of novel angiotensin I converting enzyme inhibitory dipeptides from Xerocomus badius cultured in shrimp processing waste medium. BioMed Research International. 
  5. Halim, N.R.A., Yusof, H.M. and Sarbon, N.M. (2016). Functional and bioactive properties of fish protein hydolysates and peptides: A comprehensive review. Trends in Food Science and Technology. 51: 24-33.
  6. He, S., Franco, C. and Zhang, W. (2013). Functions, applications and production of protein hydrolysates from fish processing co-products (FPCP). Food Research International. 50(1): 289-297.
  7. Kudre, T.G., Bhaskar, N. and Sakhare, P.Z. (2017). Optimization and characterization of biodiesel production from rohu (Labeo rohita) processing waste. Renewable Energy. 113: 1408-1418.
  8. Ma, T., Fu, Q., Mei, Q., Tu, Z. and Zhang, L. (2021). Extraction optimization and screening of angiotensin-converting enzyme inhibitory peptides from Channa striatus through bioaffinity ultrafiltration coupled with LC-Orbitrap-MS/MS and molecular docking. Food Chemistry. 354: 129589.
  9. Murray, B.A., Walsh, D.J. and Fitz Gerald, R.J. (2004). Modification of the furanacryloyl-L-phenylalanylglycylglycine assay for determination of angiotensin-I-converting enzyme inhibitory activity. Journal of Biochemical and Biophysical Methods. 59(2): 127-137.
  10. Naqvi, M.A., Tahir, S. and Gilani, A. (2014). Proximate composition of head and scales in wild and farmed Ctenopharyngodon idella. J. Glob. Innov. Agric. Soc. Sci. 2(4): 171-174. 
  11. Ngo, D.H., Vo, T. S., Ryu, B. and Kim, S.K. (2016). Angiotensin-I- converting enzyme (ACE) inhibitory peptides from Pacific cod skin gelatin using ultrafiltration membranes. Process Biochemistry. 51(10): 1622-1628.
  12. Poulter, N.R., Prabhakaran, D. and Caulfield, M. (2015). Hypertension. The Lancet. 386(9995): 801-812. 
  13. See, S.F., Hoo, L.L. and Babji, A.S. (2011). Optimization of enzymatic hydrolysis of Salmon (Salmo salar) skin by Alcalase. International Food Research Journal. 18(4).
  14. Skałecki, P., Florek, M., Staszowska, A. and Kaliniak, A. (2015). Use value and quality of fillets of carp fish (Cyprinidae) reared in polyculture. Żywność. Nauka. Technologia. Jakość, 1, 75-88.
  15. Theodore, A.E. and Kristinsson, H.G. (2007). Angiotensin converting enzyme inhibition of fish protein hydrolysates prepared from alkaline aided channel catfish protein isolate. Journal of the Science of Food and Agriculture. 87(12): 2353- 2357.
  16. Wasswa, J., Tang, J., Gu, X. H. and Yuan, X. Q. (2007). Influence of the extent of enzymatic hydrolysis on the functional properties of protein hydrolysate from grass carp (Ctenopharyngodon idella) skin. Food Chemistry. 104(4): 1698-1704.
  17. Wu, S., Sun, J., Tong, Z., Lan, X., Zhao, Z. and Liao, D. (2012). Optimization of hydrolysis conditions for the production of angiotensin-I converting enzyme-inhibitory peptides and isolation of a novel peptide from lizard fish (Saurida elongata) muscle protein hydrolysate. Marine drugs. 10(5): 1066-1080.
  18. Wu, W., He, L., Liang, Y., Yue, L., Peng, W., Jin, G. and Ma, M. (2019). Preparation process optimization of pig bone collagen peptide-calcium chelate using response surface methodology and its structural characterization and stability analysis. Food Chemistry. 284: 80-89.
  19. Yathisha U.G., Ishani Bhat, Iddya Karunasagar and Mamatha B.S. (2018). Antihypertensive activity of fish protein hydrolysates and its peptides. Critical Reviews in Food Science and Nutrition. 59(15): 2363-2374.
  20. Zhang, Y., Ma, L. and Otte, J. (2016). Optimization of hydrolysis conditions for production of angiotensin-converting enzyme inhibitory peptides from Basa fish skin using response surface methodology. Journal of Aquatic Food Product Technology. 25(5): 684-693.

Global Footprints