Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
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Purification of Angiogenin from Bovine Milk

우유로부터 Angiogenin의 정제

  • Nam, M.S. (Lab. of Milk Food Biochemistry, Division of Animal Science & Resources, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Bae, H.C. (Lab. of Milk Food Biochemistry, Division of Animal Science & Resources, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Park, C.S. (Research Center for Transgenic Cloned Pigs, College of Agriculture and Life Sciences, Chungnam National University)
  • 남명수 (충남대학교 농업생명과학대학 동물자원학부 유식품생물화학실) ;
  • 배형철 (충남대학교 농업생명과학대학 동물자원학부 유식품생물화학실) ;
  • 박창식 (충남대학교 농업생명과학대학 형질전환돼지연구센타)
  • Published : 2004.02.29
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Abstract

This study was carried out to establish the purification protocol of angiogenin(ANG) from bovine milk. The purification of ANG from bovine milk was performed by using cation chromatography, high-performance liquid chromatography and gel-filtration. We obtained the ANG protein have the molecular weight of about 14 kD by SDS-PAGE analysis. This protein was confirmed as ANG by Nlb-terminal sequence analysis of the first 15 amino acids. Identified amino acids revealed the protein to be identical to that previously reported for bovine ANG.

본 연구는 우유의 angiogenin(ANG) 생리활성기능을 조사하기 위한 전단계로 ANG 정제 방법을 확립하기 위하여 실시하였다. 우유로부터 ANG을 CM-Sepharose ion exchange chromatography, HPLC, Gel-filtration의 3단계 방법을 이용하여 정제하였다. 1단계인 CM-Sepharose ion exchange chromatography를 수행 한 결과 0.6 M NaCl 농도의 36번 분획에서 ANG의 밴드를 볼 수 있었고 2단계인 Mono-S 컬럼을 이용하여 HPLC를 수행한 결과에서는 1.0 M에서 하나의 peak가 분리되었고 전기영동 결과는 LF와 ANG 두 가지 성분이 함유되어 있음을 확인하였다. 마지막 단계인 Gel-filtration을 수행한 결과 ANG이 단일 밴드로 분리되었다. 우유 10 $\ell$로부터 약 80 ${\mu}\ell$의 ANG를 얻었고 정제된 ANG을 N-말단 아미노산 배열을 분석한 결과 AQDDYRYIHFLTQHY로 밝혀졌다.

Keywords

References

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