Bitterness and Solubility of Soy Protein, Casein, Gluten, and Gelatin Hydrolysates Treated with Various Enzymes

효소종류에 따른 대두단백, 카제인, 글루텐, 젤라틴 단백질 가수분해물의 쓴맛과 용해도 특성

  • 김미령 (신라대학교 바이오식품소재학과)
  • Published : 2010.04.30


To develop commercially available food protein hydrolysates, the effects of different types of enzymes and substrates on bitterness and solubility of partially hydrolyzed food proteins were investigated. Four types of proteins (casein, isolated soy protein (ISP), wheat gluten, and gelatin) and five types of proteolytic enzymes (a microbial alkaline protease (alcalase), a microbial neutral protease (neutrase), papain, bromelain, trypsin) were used. To profile the pattern of hydrolysis, the degree of hydrolysis (DH) were monitored during 180 min of reaction time by pH-stat method. Casein showed the highest susceptibility to hydrolysis for all five proteases compared to those of ISP, gluten, and gelatin. In addition, the bitter intensity and solubility (nitrogen soluble index, NSI) of each protein hydrolysate were compared at DH 10%. Bitterness and solubility of protein hydrolysates were highly affected by DH and the types of enzymes and substrates. At DH=10%, casein hydrolysate by trypsin, ISP and gluten hydrolysates by either bromelain or neutrase, and gelatin hydrolysates by the five proteases tested in this study were highly soluble and less bitter.


  1. Sosulski FW, MaCleary CW, Soliman FS. 1972. Diffusion extraction of chlorogenic acid from sunflower kernels. J Food Sci 37: 253-256.
  2. Lee CH. 1992. Application and development of protein resource. Food and Industry 25: 93-95.
  3. Ariyoshi Y. 1993. Angiotensin converting enzyme inhibitors derived from food proteins. Trends Food Sci Technol 4: 139-144.
  4. Yamamoto N. 1997. Antihypertensive peptides derived from food proteins. Biopolymers 43: 129-134.<129::AID-BIP5>3.0.CO;2-X
  5. Mesel H. 1997. Biochemical properties of bioactive peptides derived from milk proteins: potential nutraceutical for food pharmaceutical applications. Livest Prod Sci 50: 125-138.
  6. Son YS, Kwon TW. 2000. Hypocholesterolemic effect of soybean and soy products. Food Industry and Nutrition5: 36-41.
  7. Deeslie WD, Cheryan M. 1981. Continuous enzymatic modification of protein in an ultrafiltration reactor. J Food Sci 46: 1035-1042.
  8. Finley JW, Wheeler EL, Walker Jr HG, Finlayson AJ. 1982. Effect of cystine oxidation on lysinolalanine formation in proteins. J Agric Food Chem 30: 818-820.
  9. DeGroot AP, Slump P. 1969. Effects of severe alkali treatment of proteins on amino acid composition and nutritive value. J Nutr 98: 45-56.
  10. Provensal MMP, Cug JLL, Cheftel JC. 1975. Chemical and nutritional modification of sunflower proteins due to alkaline processing. Formation of amino acids crosslinks and isomerization of lysine residue. J Agric Food Chem 23: 938-943.
  11. Velisek J, Davidek T, Davidek J, Hamburg A. 1991.3-chloro-1,2-propanediol derived amino alcohol in protein hydrolysates. J Food Sci 56: 136-138.
  12. Davidek T, Davidek J, Velisek J, Kubelka V, Viden I. 1991.3-chloro-1,2-propanediol derived amino alcohol in protein hydrolysates. J Food Sci 56: 139-142.
  13. Roland JF, Mattis DL, Kiang S, Alm WL. 1978. Hydrophobic chromatography: debittering protein hydrolysates. J Food Sci 43: 1491-1493.
  14. Lee CH, Kim SK. 1987. Effects of protein hydrophobicity on the surfactant properties of food proteins. Food Hydrocolloid 1: 283-289.
  15. Adler-Nissen J. 1976. Enzymatic hydrolysis of proteins for increased solubility. J Agric Food Chem 24: 1090-1093.
  16. Lee CH, Kim CS, Lee SP. 1984. Studies on the enzymatic partial hydrolysis of soybean protein isolates. J Korean Soc Food Sci Nutr 16: 135-141.
  17. Fujimaki M, Kato H, Arai S, Tamaki E. 1968. Applying proteolytic enzymes on soybean I. Proteolytic enzyme treatment of soybean protein and its effect on the flavor. Food Tech 22: 68-77.
  18. Adler-Nissen J. 1979. Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzensulfonic acid. J Agric Food Chem 27: 1256-1262.
  19. Lin CF, Lee CR. 1987. Preparing protein for hydrolysis and product. US Patent 4,636,388.
  20. Hong YS, Lee CH, Lee KY. 1998. Effect of weak acid pretreatment on the enzymic hydrolysis against wheat gluten of high concentration. J Korean Soc Food Sci Nutr 27:1110-1116.
  21. Murray TK, Baker BE. 1952. Studies on protein hydrolysis I. Preliminary bervation on the taste of enzymatic protein hydrolyzates. J Sci Food Agric 3: 470-475.
  22. Matoba T, Hata H. 1972. Relationship between bitterness of peptides and their chemical structures. Agric Biol Chem36: 1423-1431.
  23. Fujimaki M, Kato M, Arai S, Tamaki E. 1970. Applying proteolytic enzymes on soybean 3. Diffusable bitter peptides and free amino acids in peptic hydrolyzates of soybean protein. J Food Sci 35: 215-218.
  24. Kirimura J, Shmizu A, Kimizuka A, Ninomiya T, Katsuya N. 1973. The contribution of peptides and amino acids to the taste of food stuffs. J Agric Food Chem 17: 689-695.
  25. Ney KH. 1973. Bitterness of peptide: amino acid composition and chain length. ACS Symp Ser 115: 149-173.
  26. Kim CH, Kim MR, Lee CH. 1992. Effect of type of enzyme on the bitterness of partial hydrolysates of soybean protein.Foods and Biotechnol 1: 79-84.
  27. Kim CH, Kim MR, Lee CH. 1997. The bitterness of the enzymatic hydrolysate of soybean protein and the amino acid composition of the UF filtrate. Foods and Biotechnol 6: 244-249.
  28. Jacobsen CF, Leonis J, Linderstrom-Lang K, Ottensen M. 1957. The pH-stat and its use in biochemistry. In Method of Biochemical Analysis. Interscience Publishers Inc., New York, USA. Vol 4, p 171-209.
  29. Adler-Nissen J. 1986. Enzymatic hydrolysis of proteins food proteins. Elsevier Applied Science Publishers, London and New York. p 132.
  30. Adler-Nissen J. 1984. Control of the proteolytic reaction and of the level of bitterness in protein hydrolsis processes. J Chem Tech Biotechnol 24B: 215-222.
  31. AOAC. 1980. Official methods of analysis. 13th ed. Association of Official Analytical Chemists, Washington, DC, USA. Method 2.049.
  32. AACC. 1983. Approved methods of Analysis, Crude protein-Improved Kjeldahl Methods, American Association of Cereal Chemists, St. Paul, Minnesota, USA. Method 46-10.
  33. Stryer L. 1988. Biochemistry. 3rd ed. WH Freeman and Co., New York, USA. p 178-180.
  35. Shin HS, Kim SB, Lim JW. 2002. Comparative study of proteolytic activities of some commercial milk clotting enzymes on bovine skim. J Animal Sci Technol 44: 801-808.
  36. Monti JC, Jost R. 1978. Enzymatic solublization of heat-denatured cheese whey protein. J Dairy Sci 61: 1233-1237.
  37. Li ZY, Youravong W, H-Kittikun A. 2010. Protein hydrolysis by protease isolated from tuna spleen by membrane filtration: comparative study with commercial protease.LWT-Food Sci Technol 43: 166-172.
  38. Kato A, Komatsu K, Fujimoto K. 1985. Relationship between surface functional properties and flexibility of proteins detected by the protease susceptibility. J Agric Food Chem 3: 931-934.
  39. Kong X, Zhou H, Qian H. 2006. Enzymatic preparation and functional properties of wheat gluten hydrolysates. Food Chem 101: 615-620.
  40. Sullivan JJ, Lynette M, Rood JI, Jago GR. 1973. The enzymic degradation of bitter peptides by starter Streptococci. Aust J Dairy Technol 28: 20-26.
  41. Suh HJ, Kim YS, Chung SH, Kim YS, Lee SD. 1996. Functionality and inhibitory effect of soybean hydrolysate on angiotensin converting enzyme. Korean J Food & Nutr 9: 167-175.
  42. Novo Industri A/S. 1978. Anson hemoglobin method for determination of bacterial protease activity. AF 4.2/5, Novo Industri A/S, Bagsvaerd, Denmark.

Cited by

  1. Conditions for hydrolysis of perilla seed meal protein for producing hydrolysates and ultrafiltered peptides and their antioxidant activity vol.25, pp.5, 2018,