Issue
Korean Journal of Chemical Engineering,
Vol.28, No.3, 848-852, 2011
Expression and purification of beefy meaty peptide in Pichia pastoris
We established a gene expression process to produce a flavor peptide in Pichia pastoris. The octapeptide Lys-Gly-Asp-Glu-Glu-Ser-Leu-Ala was isolated from beef digested by papain, known as “beefy meaty peptide” (BMP). It was considered a savory seasoning in previous literatures. To produce BMP by microorganisms, BMP expression structures encoding tandem repeats of the octapeptide were designed and cloned in E. coli DH5α, then integrated into the AOX1 gene of Pichia pastoris GS115. Clones containing different BMP-gene copies, with 4-copy, 8-copy, 12-copy and 16-copy, were obtained and expressed in Pichia pastoris GS115. For the ease of purification, 6×His tag was fused to the C-termini of the peptides. As a result, the fusion peptides were successfully purified by His-tag bind affinity resin. The BMP fusion peptides with expected sizes were secreted from the resulting strains of P. pastoris GS115.
[References]
  1. Yamasaki Y, Maekawa KA, Agric. Biol. Chem., 42, 1761, 1978
  2. Yamasaki Y, Maekawa K, Agric. Biol. Chem., 44, 93, 1980
  3. Wang K, Maga JA, Bechtel PJ, J. Food Sci., 61, 837, 1996
  4. Tamura M, Nakatsuka T, Tada M, Kawasaki Y, Kikuchi E, Okai H, Agric. Biol. Chem., 53, 319, 1989
  5. Wang K, Maga JA, Bechtel PJ, Lebensm.-Wiss Technol., 28, 539, 1995
  6. Sreekrishna K, Brankamp RG, Kropp KE, Blankenship DT, Tsay JT, Smith PL, Wierschke JD, Subramaniam A, Birkenberger LA, Gene., 190, 55, 1997
  7. Zhao X, Huo K, Li Y, Chinese J. Biotechnol., 16, 308, 2000
  8. Schagger H, von Jagow G, Anal. Chem., 166, 368, 1987
  9. Ellis SB, Brust PF, Koutz PJ, Waters AF, Harpold MM, Gingeras TR, Mol. Cell. Biol., 5, 1111, 1985
  10. Cregg JM, Barringer KJ, Hessler AY, Madden KR, Mol. Cell. Blol., 5, 3376, 1985
  11. Zhong XF, Peng LS, Zheng SL, Sun ZZ, Ren YF, Dong ML, Xu AL, Protein Expr. Purif., 36(2), 165, 2004
  12. Xie JL, Zhang L, Ye Q, Zhou QW, Xin L, Du P, Gan RB, Biotechnol. Lett., 25(2), 173, 2003
  13. Chung HJ, Park SM, Kim HR, Yang MS, Kim DH, Enzyme Microb. Technol., 31(4), 384, 2002
  14. Nakata T, Takahashi M, Nakatani M, Kuramitsu R, Tamura M, Okai H, Japan Society for Bioscience, Biotechnology, and Agrichemistry., 59, 689, 1995
  15. Moralejo FJ, Cardoza RE, Gutierrez S, Martin JF, Appl. Environ. Microbiol., 65, 1168, 1999
  16. Kondo K, Miura Y, Sone H, Kobayashi K, Iijima H, Nat. Biotechnol., 15, 453, 1997