Detection of the Cholera Toxin-binding Activity of κ-Casein Macropeptide and Optimization of Its Production by the Response Surface Methodology

Sejong Oh, Randy W. Worobo, Byoung-Chul Kim, Sungsue Rheem, Sae Hun Kim

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The cholera toxin (CT)-binding activity of purified κ-casein macropeptide (CMP) from bovine κ-casein was detected. In addition, a statistical model was developed to optimize the production of CMP. CMP was prepared by chymosin hydrolysis of κ-casein and a subsequent 3% trichloroacetic acid treatment. CMP was further fractionated in an ion-exchange column by FPLC. CT binding activity was eluted at 0.18 M NaCl and was a single 8.9 kDa peptide without tyrosine and arginine residues. The CT binding activity was rapidly lost by a carbohydrase treatment. The conditions for CMP production with chymosin were optimized by using the response surface methodology (RSM). The estimated optimum levels of the factors were as follows: reaction temperature, 38.5°C; pH, 6.44; and time, 35.9 min. A validation experiment was performed in which CMP was prepared under the predicted parameters, and it was ascertained that the estimated optimum conditions gave better production of CMP than any other conditions.

Original languageEnglish
Pages (from-to)516-522
Number of pages7
JournalBioscience, Biotechnology and Biochemistry
Volume64
Issue number3
Publication statusPublished - 2000 Mar 1

Fingerprint

Casein
cholera toxin
Cholera Toxin
Caseins
response surface methodology
casein
Chymosin
chymosin
Trichloroacetic acid
Trichloroacetic Acid
Arginine
trichloroacetic acid
Ion Exchange
ion exchange
Statistical Models
acid treatment
statistical models
Peptides
arginine
Tyrosine

Keywords

  • κ-casein macropeptide
  • Cholera toxin
  • Response surface methodology

ASJC Scopus subject areas

  • Food Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Chemistry (miscellaneous)
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

Detection of the Cholera Toxin-binding Activity of κ-Casein Macropeptide and Optimization of Its Production by the Response Surface Methodology. / Oh, Sejong; Worobo, Randy W.; Kim, Byoung-Chul; Rheem, Sungsue; Kim, Sae Hun.

In: Bioscience, Biotechnology and Biochemistry, Vol. 64, No. 3, 01.03.2000, p. 516-522.

Research output: Contribution to journalArticle

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