Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterizating of Recombinant Human Albumin from Hansenula polymorpha DL-1

Hansenula polymorpha DL-1이 생산하는 재조합 알부민의 정제 및 특성

  • 최근범 (동국제약 (주) 중앙연구소) ;
  • 구선향 (동국제약 (주) 중앙연구소) ;
  • 임채양 (동국제약 (주) 중앙연구소) ;
  • 이동희 (건국대학교 미생물공학과) ;
  • 강현아 (동국제약 (주) 중앙연구소) ;
  • 이상기 (생명공학연구원)
  • Published : 2001.12.01
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Abstract

Recombinant Human serum albumin (rHSA) was purified to near homogeneity from H, polymorpha using heat treatment, ultrafiltratipn and Phenyl Sepharose CL-4B and Mono Q column chro - matographies with a recovery yield of 60% The molecular weight of the purified rHSA was estimated to be about 65,000 Da by denaturing SDS-PAGE The N terminal amino acid sequence of the purified HSA determined by Edman degradation was turned out to be Asp- Ala- His- Lys- Ser- Glu- Ala, suggesting that the rHSA expressed in H, polymorpha was efficiently secreted and correctly processed at the cleavage site of secretion signal sequence. The purified human albumin showed the pI value identical to that of authentic human serum albumin.

메탄올 자화효모 Hansenula polymorpha 가 생산하는 재조합 인체 알부민 (human albumin)을 heat treatment ultrafiltration phenyl Sepharose CL-4B Mono Q 컬럼 크로마토그래피를 수행하여 수율 60%로 정제하였다. 정제된 재보합 알부민 SDS-PAGE를 수행한 결과 분자량이 65,000 Da으로 나타났고, 본 재조합 알부민의 N-말단 아미노산 서열은 Asp- Ala- His- Lys- Ser- Glu- Ala 으로 혈청유래 알부민 및 다른 효모 유래의 재조합 인체 알부민 과 동일함을 보였다. 정제한 재조합 인체 알부민의 아미노산 조성은 총 18종 의 아미노산이 검출되었고 아미노산 잔기 중 cysteine, arginine lysine 등이 이론치와 일치하였으며, 혈청 알부민과 동일하게 약 pI 4.8 정도 값을 나타내어 H. polymorpha 유래의 재조합 단백질의 전반적인 아미노산 구성이 혈청 알부민과 동일함을 확인하였다.

Keywords

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