서지주요정보
금속 친화성 트로마토그래피를 이용한 항혈전 단백질 히루딘의 분리 및 정제 = Sepatation and purification of anticoagulant hirudin by metal affinity chromatography
서명 / 저자 금속 친화성 트로마토그래피를 이용한 항혈전 단백질 히루딘의 분리 및 정제 = Sepatation and purification of anticoagulant hirudin by metal affinity chromatography / 추창웅.
저자명 추창웅 ; Chu, Chang-Woong
발행사항 [대전 : 한국과학기술원, 1993].
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소장정보

등록번호

8003888

소장위치/청구기호

학술문화관(문화관) 보존서고

MCHE 93036

휴대폰 전송

도서상태

이용가능

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반납예정일

초록정보

In this study, hirudin, a protease inhibitor from the salivary gland of bloodsucking leech, was purified by immobilized metal affinity chromatography. Divalent metal ions such as copper, nickel, and zinc were used as ligand. Metal ions were immobilized on the gel, sepharose and then chelated with hirudin. The binding strength between metal ion and hirudin was dependent on the secondary structure of the chelation site on hirudin. Several types of hirudin were prepared by gene cloning of yeast, Sacaromyces cerevisiae. Four wild type hirudin were obtained : wild type, MH7, MH9, and MH13. The wild type hirudin has only one histidine residue at 51st site in its amino acid sequence. Hirudin MH7 has two histidine residues at 51st and 48th sites, while MH9 at 51st and 54th sites. MH13 has three histidine residues at 48th, 51st, and 54th sites. The metal ion affinity of hirudin variants were tested. With copper ion all hirudin variants had affinity, and they were eluted after the pH gradient were started. The wild type hirudin was eluted at pH 5.5, MH7 at pH 4.4, MH9 at pH 5.2, MH13 type at pH 4.0. Thus, it was concluded that MH13 type had the strongest affinity of all hirudin variants. The specific activity of the wild type hirudin is 13,000 ATU/mg. But, those of the others were lower than this ; MH7 : 11,000 ATU/mg; MH9 : 9,700 ATU/mg; MH13 : 4,300 ATU/mg. As more histidines were substituted, the specific activity of the hirudin variants decreased. The adsorption mechanism of wild type hirudin was studied and a mathematical model. The surface reaction constant was so large that the adsorption reaction at the surface was considered very fast. Since the coefficient for mass transfer through the liquid film was quite small, this step was considered to be the rate-limiting step. Several purification processes for the wild type and MH7 hirudins were tested. Each process was composed of an ultrafiltration with molecular weight cut-off of 30,000 an ultrafiltration with molecular weight cut-off of 3,000 an anion exchange chromatography and a copper ion metal ion affinity chromatography. Descriptions on the performance of each process has been presented in detail.

서지기타정보

서지기타정보
청구기호 {MCHE 93036
형태사항 xi, 82 p. : 삽도 ; 26 cm
언어 한국어
일반주기 저자명의 영문표기 : Chang-Woong Chu
지도교수의 한글표기 : 장용근
지도교수의 영문표기 : Yong-Keun Chang
학위논문 학위논문(석사) - 한국과학기술원 : 화학공학과,
서지주기 참고문헌 : p. 80-82
주제 Separation (Technology)
Chromatographic analysis.
분리. --과학기술용어시소러스
크로마토 그래피. --과학기술용어시소러스
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