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Current IssueEditorial BoardOnline First ArticlesArchive

volume 38 issue 1 (february 2015) : 60-65,   Doi: 10.5958/0976-0571.2015.00010.7

Genetic variability and effect of heat treatment on trypsin inhibitor content in soybean [Glycine max (L.) Merrill.]

M
M. Shivakumar
K
Khushbu Verma
A
Akshay Talukdar*
N
Nidhi Srivastava
S
S.K. Lal
R
R.L. Sapra
K
K.P. Singh
1Division of Genetics, Indian Agricultural Research Institute, New Delhi-110 012, India.
Cite article:- Shivakumar M., Verma Khushbu, Talukdar* Akshay, Srivastava Nidhi, Lal S.K., Sapra R.L., Singh K.P. (2025). Genetic variability and effect of heat treatment on trypsin inhibitor content in soybean [Glycine max (L.) Merrill.]. Legume Research. 38(1): 60-65. doi: 10.5958/0976-0571.2015.00010.7.

ABSTRACT

Kunitz trypsin inhibitor (KTI), a major anti-nutritional factor present in soybean seed, is a major bottleneck in industrial use of soybean and public acceptance of soybean food products. Biochemical screening for trypsin inhibitor content was carried out in 145 genotypes of soybean comprising exotic collections and released varieties. The trypsin inhibitor content ranged from 14.15 to 186.27 mg g-1 of seed meal. Lowest level of trypsin inhibitor content was observed in genotype PI542044, (14.15 mg g-1 seed meal). Popular Indian soybean varieties viz., JS335, JS9305, NRC37, NRC7, PK1225, DS9712, JS9752, and DS9814 contained higher level of trypsin inhibitor. Heat treatment was able to reduce the TI content but to certain level leaving about 20% residual activity. The study indicated the need for other ways to create genotypes with reduced KTI for use in breeding program.

REFERENCES

  1. Anderson R. L. (1992). Effect of steaming on soybean proteins and trypsin inhibitors. J. Am. Oil Chem. Soc., 69: 1170-1176.
  2. Armstrong W.B. Kennedy A.R. Wan X.S. Atiba J. Mclaren E. and Meyskens F.L (2000). Single-dose administration of Bowmen-Birk inhibitor concentrate in patients with oral leukoplakia. Cancer Epidemiolgy Biomark Prev., 9: 43-47.
  3. Bau H.M. Villaume C. Giannangeli F. Nicolas J.P. and Mejean L. (2001). Optimisation du chauffage et valeurs nutritionnelle et fonctionnelle des prote´ines de soja. Cahiers de nutrition et de die´te´tique., 36:96–102.
  4. Dipika A. Murugkar and Krishna J. (2010). Effect of drying on nutritional and functional quality and electrophoretic pattern of soyflour from sprouted soybean (G. max). J. Food sci. Technol., 47(5):482-487.
  5. Evandro F. F. Jack H. W. and Tzi B.N. (2010). Thermostable kunitz trypsin inhibitor with cytokine inducing, antitumor and HIV-1 reverse transcriptase inhibitory activities from Korean large black soybeans. Journal of Bioscience and Bioengineering., 109(3): 211–217.
  6. Gran B. Tabibzadeh N. Martin A. Ventura E.S. Ware J.H. Zhang G.X. Parr J.L. Kennedy A.R. Rostami A.M. (2006). The protease inhibitor, Bowman-Birk inhibitor, suppresses experimental autoimmune encephalomyelitis: a potential oral therapy for multiple sclerosis. Mult. Scler. J., 12: (6) 688-697.
  7. Hamerstrand G. E. Black L.T. and Glover J.D. (1981). Trypsin Inhibitors in Soy products: Modification of the Standard Analytical Procedure. Cereal Chemistry., 58(1):42-45.
  8. Huma H. and Khalid M.F. (2007). Plant protease inhibitors: a defense strategy in plants. Biotechnology and Molecular Biology Review., 2(3): 068-085
  9. Hymowitz T. (1973). Electrophoretic analysis of SBTI-A2 in the USFA soybean germplasm collection. Crop Sci., 13:420–421.
  10. Jofuku D.K. Roberta D. Schipper and Robert B. Goldberg. (1989). A Frameshift mutation prevents kunitz trypsin inhibitor mRNA accumulation in soybean embryos. The Plant Cell., 1:427-435.
  11. Kim M.S Park M.J Jeong W.H. Nam K.C. Chung J.I. (2006). SSR marker tightly linked to the Ti locus in soybean [Glycine max (L.) Merr.]. Euphytica., 152: 361-366.
  12. Krishnan H. B. Kim W.S. (2003). A four-nucleotide base-pair deletion in the coding region of the Bowman–Birk protease inhibitor gene prevents its accumulation in the seeds of Glycine microphylla PI440956. Planta., 217: 523–527.
  13. Krishnan H.B. (2001). Characterization of a soybean [Glycine max (L.) Merr.] mutant with reduced levels of Kunitz trypsin inhibitor. Plant Science., 160: 979–986.
  14. Kumar V, Rani A., Maurya V., Rawal R., Verma K., Shivamumar M.,Lal, SK., and Talukdar Akshay. (2011). Marker-assisted selection for development of kunitz trypsin inhibitor free soybean varieties: I: parental polymorphism survey using SSR markers. Indian J. Genet. 70(4): 372-376
  15. Kumar V. Rani A. and Tiwari S.P. (2001). Comparative activity of trypsin inhibitor among released soybean varieties/strains of India. Ind. J. of Nut. Diet., 38: 437-440.
  16. Kumar V. Rani A. Bollore S.D. and Chauhan G.S. (2006). Physico-chemical properties of immature pods of Japanese soybean cultivars. International Journal of Food Properties. 9: 51-59.
  17. Leontowicz H. Kostyra H. Leontowicz M. and Kulasek G.W. (1998). The inactivation of legume seed haemagglutinin and trypsin inhibitors by boiling. In (Eds.), Recent advances of research in antinutritional factors in legume seeds and rapeseed (pp. 429–    432). The Netherlands: Wageningen Press.
  18. Liener I. E. (1994). Implications of antinutritional components in soybean foods. 34: 31-37.
  19. Liener I. E. and Kakade M. L. (1980). Protease inhibitors. In: Toxic-Constituents of Plant Foodstuffs, 2nd ed (ed. I. E. Liener). Academic Press, New York: 7-71.
  20. Machado F.P.P. Queiro J.H. Oliveira M.G.A. Piovesan N.D. Peluzio M.C.G. Costa N.M.B. and Moreira M.A. (2008). Effects of heating on protein quality of soybean flour devoid of Kunitz inhibitor and lectin. Food Chem., 107: 649–655.
  21. Manjaya T. Gopalakrishna S.E. Pawar and Bapat V.A. (2007). Genetic variability for trypsin inhibitor content in soybean [Glycine max (L.) Merrill.] and its correlation with oil and protein. Indian J. Genet., 67(1): 51-55.
  22. Michael T.M. William A.L. Lyn M.W. Ngaire M. Christine R.V. Derek W.R. and White. (2005). Expression of the soybean (Kunitz) trypsin inhibitor in leaves of white clover (Trifolium repens L.). Plant Science. 168: 1211–1220.
  23. Orf J.H. and Hymowitz T. (1979). Inheritance of the absence of the Kunitz trypsin inhibitor in seed protein of soybeans. Crop Sci., 19:107–109.
  24. Rani A. Kumar V. Mourya V. Singh R.K. and Husain S.M. (2011). Validation of SSR markers linked to null kunitz trypsin inhibitor allele in Indian soybean [Glycine max (L.) Merr.] population. J. Plant Biochem. Biotechnol., 20(2):258-261.
  25. Rios-Iriarte B. J. and Barnes R. H. (1996). The effect of overheating on certain nutritional properties of the proteins of soybean. Food Technology., 32: 836–839.
  26. Singh L.C. Wilson M. and Hadley H.H. (1969). Genetic differences in soybean trypsin inhibitor separated by disc electrophoresis. Crop Sci., 9:489–491.
  27. Wang K.J. Kaizuma N. Takahata Y. and Hatakeyama S. (1996). Detection of two new variants of soybean Kunitz trypsin inhibitor through electrophoresis. Breed Sci., 46:39–44.
  28. Wang K.J. and Li X.H. (2005). Tif type of soybean Kunitz trypsin inhibitor exists in wild soybean of northern China. In: Proceedings of the 8th national soybean research conference of China, pp 167–168.
  29. Wang K.J. Takahata Y. Ito K. Zhao Y.P. Tsutsumi K.I. and Kaizuma N. (2001). Genetic characterization of a novel soybean kunitz trypsin inhibitor. Breed. Sci.51:185–190.
  30. Wang K.J. Takahata Y. Kono Y. and Kaizuma N. (2008). Allelic differentiation of Kunitz trypsin inhibitor in wild soybean. (Glycine soja). Theor. Appl. Genet., 117:565–573.
  31. Wang K.J. Xiang H.L. Yamashita T. and Yoshihito T. (2012). Single nucleotide mutation leading to an amino acid substitution in the variant Tik soybean Kunitz trypsin inhibitor (SKTI) identified in Chinese wild soybean (Glycine soja Sieb. & Zucc.). Plant Syst Evol., 298:1–7.
  32. Wang K.J. Yamashita T. Watanabe M. and Yoshihito T. (2004). Genetic characterization of a novel Tib-derived variant of soybean Kunitz trypsin inhibitor detected in wild soybean (Glycine soja). Genome., 47: 9–14.
  33. Zhao S.W. and Wang H. (1992). A new electrophoretic variant of SBTi-A2 in soybean seed protein. Soybean Genet. Newslett., 19:22–24.
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    volume 38 issue 1 (february 2015) : 60-65,   Doi: 10.5958/0976-0571.2015.00010.7

    Genetic variability and effect of heat treatment on trypsin inhibitor content in soybean [Glycine max (L.) Merrill.]

    M
    M. Shivakumar
    K
    Khushbu Verma
    A
    Akshay Talukdar*
    N
    Nidhi Srivastava
    S
    S.K. Lal
    R
    R.L. Sapra
    K
    K.P. Singh
    1Division of Genetics, Indian Agricultural Research Institute, New Delhi-110 012, India.
    Cite article:- Shivakumar M., Verma Khushbu, Talukdar* Akshay, Srivastava Nidhi, Lal S.K., Sapra R.L., Singh K.P. (2025). Genetic variability and effect of heat treatment on trypsin inhibitor content in soybean [Glycine max (L.) Merrill.]. Legume Research. 38(1): 60-65. doi: 10.5958/0976-0571.2015.00010.7.

    ABSTRACT

    Kunitz trypsin inhibitor (KTI), a major anti-nutritional factor present in soybean seed, is a major bottleneck in industrial use of soybean and public acceptance of soybean food products. Biochemical screening for trypsin inhibitor content was carried out in 145 genotypes of soybean comprising exotic collections and released varieties. The trypsin inhibitor content ranged from 14.15 to 186.27 mg g-1 of seed meal. Lowest level of trypsin inhibitor content was observed in genotype PI542044, (14.15 mg g-1 seed meal). Popular Indian soybean varieties viz., JS335, JS9305, NRC37, NRC7, PK1225, DS9712, JS9752, and DS9814 contained higher level of trypsin inhibitor. Heat treatment was able to reduce the TI content but to certain level leaving about 20% residual activity. The study indicated the need for other ways to create genotypes with reduced KTI for use in breeding program.

    REFERENCES

    1. Anderson R. L. (1992). Effect of steaming on soybean proteins and trypsin inhibitors. J. Am. Oil Chem. Soc., 69: 1170-1176.
    2. Armstrong W.B. Kennedy A.R. Wan X.S. Atiba J. Mclaren E. and Meyskens F.L (2000). Single-dose administration of Bowmen-Birk inhibitor concentrate in patients with oral leukoplakia. Cancer Epidemiolgy Biomark Prev., 9: 43-47.
    3. Bau H.M. Villaume C. Giannangeli F. Nicolas J.P. and Mejean L. (2001). Optimisation du chauffage et valeurs nutritionnelle et fonctionnelle des prote´ines de soja. Cahiers de nutrition et de die´te´tique., 36:96–102.
    4. Dipika A. Murugkar and Krishna J. (2010). Effect of drying on nutritional and functional quality and electrophoretic pattern of soyflour from sprouted soybean (G. max). J. Food sci. Technol., 47(5):482-487.
    5. Evandro F. F. Jack H. W. and Tzi B.N. (2010). Thermostable kunitz trypsin inhibitor with cytokine inducing, antitumor and HIV-1 reverse transcriptase inhibitory activities from Korean large black soybeans. Journal of Bioscience and Bioengineering., 109(3): 211–217.
    6. Gran B. Tabibzadeh N. Martin A. Ventura E.S. Ware J.H. Zhang G.X. Parr J.L. Kennedy A.R. Rostami A.M. (2006). The protease inhibitor, Bowman-Birk inhibitor, suppresses experimental autoimmune encephalomyelitis: a potential oral therapy for multiple sclerosis. Mult. Scler. J., 12: (6) 688-697.
    7. Hamerstrand G. E. Black L.T. and Glover J.D. (1981). Trypsin Inhibitors in Soy products: Modification of the Standard Analytical Procedure. Cereal Chemistry., 58(1):42-45.
    8. Huma H. and Khalid M.F. (2007). Plant protease inhibitors: a defense strategy in plants. Biotechnology and Molecular Biology Review., 2(3): 068-085
    9. Hymowitz T. (1973). Electrophoretic analysis of SBTI-A2 in the USFA soybean germplasm collection. Crop Sci., 13:420–421.
    10. Jofuku D.K. Roberta D. Schipper and Robert B. Goldberg. (1989). A Frameshift mutation prevents kunitz trypsin inhibitor mRNA accumulation in soybean embryos. The Plant Cell., 1:427-435.
    11. Kim M.S Park M.J Jeong W.H. Nam K.C. Chung J.I. (2006). SSR marker tightly linked to the Ti locus in soybean [Glycine max (L.) Merr.]. Euphytica., 152: 361-366.
    12. Krishnan H. B. Kim W.S. (2003). A four-nucleotide base-pair deletion in the coding region of the Bowman–Birk protease inhibitor gene prevents its accumulation in the seeds of Glycine microphylla PI440956. Planta., 217: 523–527.
    13. Krishnan H.B. (2001). Characterization of a soybean [Glycine max (L.) Merr.] mutant with reduced levels of Kunitz trypsin inhibitor. Plant Science., 160: 979–986.
    14. Kumar V, Rani A., Maurya V., Rawal R., Verma K., Shivamumar M.,Lal, SK., and Talukdar Akshay. (2011). Marker-assisted selection for development of kunitz trypsin inhibitor free soybean varieties: I: parental polymorphism survey using SSR markers. Indian J. Genet. 70(4): 372-376
    15. Kumar V. Rani A. and Tiwari S.P. (2001). Comparative activity of trypsin inhibitor among released soybean varieties/strains of India. Ind. J. of Nut. Diet., 38: 437-440.
    16. Kumar V. Rani A. Bollore S.D. and Chauhan G.S. (2006). Physico-chemical properties of immature pods of Japanese soybean cultivars. International Journal of Food Properties. 9: 51-59.
    17. Leontowicz H. Kostyra H. Leontowicz M. and Kulasek G.W. (1998). The inactivation of legume seed haemagglutinin and trypsin inhibitors by boiling. In (Eds.), Recent advances of research in antinutritional factors in legume seeds and rapeseed (pp. 429–    432). The Netherlands: Wageningen Press.
    18. Liener I. E. (1994). Implications of antinutritional components in soybean foods. 34: 31-37.
    19. Liener I. E. and Kakade M. L. (1980). Protease inhibitors. In: Toxic-Constituents of Plant Foodstuffs, 2nd ed (ed. I. E. Liener). Academic Press, New York: 7-71.
    20. Machado F.P.P. Queiro J.H. Oliveira M.G.A. Piovesan N.D. Peluzio M.C.G. Costa N.M.B. and Moreira M.A. (2008). Effects of heating on protein quality of soybean flour devoid of Kunitz inhibitor and lectin. Food Chem., 107: 649–655.
    21. Manjaya T. Gopalakrishna S.E. Pawar and Bapat V.A. (2007). Genetic variability for trypsin inhibitor content in soybean [Glycine max (L.) Merrill.] and its correlation with oil and protein. Indian J. Genet., 67(1): 51-55.
    22. Michael T.M. William A.L. Lyn M.W. Ngaire M. Christine R.V. Derek W.R. and White. (2005). Expression of the soybean (Kunitz) trypsin inhibitor in leaves of white clover (Trifolium repens L.). Plant Science. 168: 1211–1220.
    23. Orf J.H. and Hymowitz T. (1979). Inheritance of the absence of the Kunitz trypsin inhibitor in seed protein of soybeans. Crop Sci., 19:107–109.
    24. Rani A. Kumar V. Mourya V. Singh R.K. and Husain S.M. (2011). Validation of SSR markers linked to null kunitz trypsin inhibitor allele in Indian soybean [Glycine max (L.) Merr.] population. J. Plant Biochem. Biotechnol., 20(2):258-261.
    25. Rios-Iriarte B. J. and Barnes R. H. (1996). The effect of overheating on certain nutritional properties of the proteins of soybean. Food Technology., 32: 836–839.
    26. Singh L.C. Wilson M. and Hadley H.H. (1969). Genetic differences in soybean trypsin inhibitor separated by disc electrophoresis. Crop Sci., 9:489–491.
    27. Wang K.J. Kaizuma N. Takahata Y. and Hatakeyama S. (1996). Detection of two new variants of soybean Kunitz trypsin inhibitor through electrophoresis. Breed Sci., 46:39–44.
    28. Wang K.J. and Li X.H. (2005). Tif type of soybean Kunitz trypsin inhibitor exists in wild soybean of northern China. In: Proceedings of the 8th national soybean research conference of China, pp 167–168.
    29. Wang K.J. Takahata Y. Ito K. Zhao Y.P. Tsutsumi K.I. and Kaizuma N. (2001). Genetic characterization of a novel soybean kunitz trypsin inhibitor. Breed. Sci.51:185–190.
    30. Wang K.J. Takahata Y. Kono Y. and Kaizuma N. (2008). Allelic differentiation of Kunitz trypsin inhibitor in wild soybean. (Glycine soja). Theor. Appl. Genet., 117:565–573.
    31. Wang K.J. Xiang H.L. Yamashita T. and Yoshihito T. (2012). Single nucleotide mutation leading to an amino acid substitution in the variant Tik soybean Kunitz trypsin inhibitor (SKTI) identified in Chinese wild soybean (Glycine soja Sieb. & Zucc.). Plant Syst Evol., 298:1–7.
    32. Wang K.J. Yamashita T. Watanabe M. and Yoshihito T. (2004). Genetic characterization of a novel Tib-derived variant of soybean Kunitz trypsin inhibitor detected in wild soybean (Glycine soja). Genome., 47: 9–14.
    33. Zhao S.W. and Wang H. (1992). A new electrophoretic variant of SBTi-A2 in soybean seed protein. Soybean Genet. Newslett., 19:22–24.
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