Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Development of Methods for Protein Extraction from Three Major Korean Fermented Soy Foods for 2-Dimensional Gel and Mass Spectrometric Analyses

  • Lim, Jin-Kyu (Department of Animal Science and Biotechnology, Kyungpook National University)
  • Published : 2008.06.30
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Abstract

Three different protein extraction methods-phenol extraction, trichloroacetic acid (TCA) precipitation, and desalting/TCA precipitation-were compared to determine the optimal reproducible high resolution 2-dimensional (2-D) electrophoresis for each chungkugjang, doenjang, and kochujang samples. The soluble proteins from Chungkugjang extracted by phenol were separated with high reproducibility and resolution, and gained 1.75- to 3-fold more protein spots on 2-D gel than those from the other methods. On the contrary, the extracted proteins from doenjang and kochujang treated by desalting/TCA precipitation method showed about 1.5- to 3.3-fold more protein spots on 2-D gel. Using the established methods, the changes in the protein profiles of the fermented soy foods were monitored during the fermentation period by 2-DE. One of the major proteins in soy, $\beta$-conglycinin $\alpha$-subuint, and some proteins with unknown functions were localized on 2-D gel as the protease-resistant proteins throughout the fermentation period of doenjang. Changes in the protein profile monitored by the established methods can provide basic information on unfolding the mechanisms of the generation of biofunctional activity in the fermented soy foods.

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References

  1. Cottrell JS (1994) Protein identification by peptide mass fingerprinting. Pept Res 7, 115-124
  2. Gianazza E, Eberini I, Arnoldi A, Wait R, and Sirtori CR (2003) A proteomic investigation of isolated soy proteins with variable effects in experimental and clinical studies. J Nutr 133, 9-14
  3. Im MH, Choi JD, Chung HC, Lee SH, Lee CW, Choi C, and Choi KS (1998) Improvement of Meju preparation method for the production of Korean traditional Kanjang (soy sauce). Korean J Food Sci Technol 30, 608-614
  4. Jiang L, He L, and Fountoulakis M (2004) Comparison of protein precipitation methods for sample preparation prior to proteomic analysis. J Chromatogr A 1023, 317-320 https://doi.org/10.1016/j.chroma.2003.10.029
  5. Kang SG, Park IB, and Jung ST (1997) Characteristics of fermented hot pepper soybean paste (Kochujang) prepared by liquid Beni-koji. Korean J Food Sci Technol 29, 82-89
  6. Kim CH, Sumino T, Aida K, Kaneko K, Yamada K, and Knead T (1993) Sodium and potassium contents of Doenjang (bean paste) made in Korean and Japan. Korean J Dietary Culture 8, 73-77
  7. Kim DH, Yang SE, and Rhim JH (2003) Fermentation characteristics of Kochujang prepared with various salts. Korean J Food Sci Technol 35, 6771-6779
  8. Lee KH, Lee HJ, and Jung MK (1971) Studies on Chunggukjang- on the changes of soy-bean protein in manufacturing Chunggukjang. J Korean Agr Chem Soc 14, 191-200
  9. Lovati M, Manzoni C, Gianazza E, Arnoldi A, Kurowska E, Carroll KK, and Sirtori CR (2000) Soy protein peptides regulate cholesterol homeostasis in Hep G2 cells. J Nutr 130, 2543-2549
  10. Mooney BP, Krishnan HB, and Thelen JJ (2004) Highthroughput peptide mass fingerprinting of soybean seed proteins: automated workflow and utility of UniGene expressed sequence tag databases for protein identification. Phytochemistry 65, 1733-1734 https://doi.org/10.1016/j.phytochem.2004.04.011
  11. Natarajan S, Xu C, Bae H, Bailey BA, Cregan P, Caperna TJ, Garrett WM, and Luthria D (2007) Proteomic and genetic analysis of glycinin subunits of sixteen soybean genotypes. Plant Physiol Biochem 45, 436-444 https://doi.org/10.1016/j.plaphy.2007.03.031
  12. Natarajan S, Xu C, Caperna TJ, and Garrett WM (2005) Comparison of protein solubilization methods suitable for proteomic analysis of soybean seed proteins. Anal Biochem 342, 214-220 https://doi.org/10.1016/j.ab.2005.04.046
  13. Oh J, Pyo JH, Jo EH, Hwang SI, Kang SC, Jung JH, Park EK, Kim SY, Choi JY, and Lim J (2004) Establishment of a near-standard two-dimensional human urine proteomic map. Proteomics 4, 3485-3497 https://doi.org/10.1002/pmic.200401018
  14. Pappin DJ, Hojrup P, and Bleasby AJ (1993) Rapid identification of proteins by peptide-mass fingerprinting. Curr Biol 3, 327-332 https://doi.org/10.1016/0960-9822(93)90195-T
  15. Park KY, Hwang MK, Jung KO, and Lee KB (2002) Studies on the standardization of Doenjang (Korean soybean paste) 1. Standardization of manufacturing method of Doenjang by literatures. J Korean Soc Food Sci Nutr 31, 343-350 https://doi.org/10.3746/jkfn.2002.31.2.343
  16. Pyo J, Hwang SI, Oh J, Lee SJ, Kang SC, Kim JS, and Lim J (2003) Characterization of a bovine pregnancyassociated protein using two-dimensional gel electrophoresis, N-terminal sequencing and mass spectrometry. Proteomics 3, 2420-2427 https://doi.org/10.1002/pmic.200300618
  17. Santos I, Sohn IY, Choi HS, Park SM, Ryu SH, Kwon DY, Park C, Kim JH, Kim JS, and Lim J (2007) Changes of protein profiles in Chunggukjang during the fermentation period. Korean J Food Sci Technol 39, 438-446
  18. Seok YR, Kim YH, Kim S, Woo HS, Kim TW, Lee SH, and Choi C (1994) Change of protein and amino acid composition during Chungkook-Jang fermentation using Bacillus licheniformis CN-115. Agr Chem Biotech 37, 65-71
  19. Sirtori CR, Even R, and Lovati MR (1993) Soybean protein diet and plasma cholesterol: from therapy to molecular mechanisms. Ann NY Acad Sci 676, 188-201 https://doi.org/10.1111/j.1749-6632.1993.tb38734.x
  20. Thanh VH and Shibasaki KJ (1977) Beta-conglycinin from soybean proteins. Isolation and immunological and physicochemical properties of the monomeric forms. Biochim Biophys Acta 26, 692-695
  21. Wilkins MR, Sanchez JC, Gooley AA, Appel RD, Humphery-Smith I, Hochstrasser DF, and Williams KL (1995) Progress with proteome projects: why all proteins expressed by a genome should be identified and how to do it. Biotech Gene Eng Rev 13, 19-50
  22. Yates III JR, Speicher S, Griffin PR, and Hunkapiller T (1993) Peptide mass maps: a highly informative approach to protein identification. Anal Biochem 214, 397-408 https://doi.org/10.1006/abio.1993.1514
  23. Yeo YK and Kim ZU (1978) Studies on the standardization of the processing condition of KoChooJang (red pepper paste). J Korean Agricultural Chem Soc 21, 16-21

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