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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제18권 제5호
- 발행연도
- 2008.1
- 수록면
- 983 - 989 (7page)
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초록· 키워드
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Human insulin is a hormone well-known to regulate the blood glucose level. Recombinant preproinsulin, a precursor of authentic insulin, is typically produced in E. coli as an inactive inclusion body, the solubilization of which needs the addition of reducing agents such as β-mercaptoethanol.
To make authentic insulin, recombinant preproinsulin is modified enzymatically by trypsin and carboxypeptidase B. The effects of β-mercaptoethanol on the formation of human insulin derivatives were investigated in the enzymatic modification by using commercially available human proinsulin as a substrate. Addition of 1 mM β- mercaptoethanol induced the formation of various insulin derivatives. Among them, the second major one, impurity 3, was found to be identical to the insulin B chain fragment from Phe1 to Glu21. Minimization of the formation of insulin derivatives and concomitant improvement of the production yield of human insulin were achieved by the addition of hydrogen peroxide. Hydrogen peroxide bound with β-mercaptoethanol and thereby reduced the negative effects of β-mercaptoethanol considerably. Elimination of the impurity 3 and other derivatives by the addition of over 10 mM hydrogen peroxide in the presence of β- mercaptoethanol led to a 1.3-fold increase in the recovery efficiency of insulin, compared with those for the case without hydrogen peroxide. The positive effects of hydrogen peroxide were also confirmed with recombinant human preproinsulin expressed in recombinant E. coli as an inclusion body.
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참고문헌 (20)
참고문헌 신청
Castellanos-Serra, L. R. / 1996 / Expression and folding of an interleukin-2-proinsulin fusion protein and its conversion into insulin by a single step enzymatic removal of t
Chen, J. Q. / 1995 / Production of human insulin in an Escherichia coli system with Met-Lys-human proinsulin as the expressed precursor. Appl. Biochem / Biotechnol 55 : 5 ~ 1
Fischer, B. / 1993 / Isolation, renaturation, and formation of disulfide bonds of eukaryotic proteins expressed in Escherichia coli as inclusion bodies. Biotechnol / Bioeng 4
Jonasson, P. / 1996 / Single-step trypsin cleavage of a fusion protein to obtain human insulin and its C peptide. Eur / Biochem 236 : 656 ~ 661
KOO, TAI YOUNG / 2007 / Expression of Recombinant Human Growth Hormone in a Soluble Form inEscherichia coli by Slowing Down the Protein Synthesis Rate / Journal of Microbiolo
Chen, J. Q. / 1995 / Production of human insulin in an Escherichia coli system with Met-Lys-human proinsulin as the expressed precursor. Appl. Biochem / Biotechnol 55 : 5 ~ 1
Fischer, B. / 1993 / Isolation, renaturation, and formation of disulfide bonds of eukaryotic proteins expressed in Escherichia coli as inclusion bodies. Biotechnol / Bioeng 4
Jonasson, P. / 1996 / Single-step trypsin cleavage of a fusion protein to obtain human insulin and its C peptide. Eur / Biochem 236 : 656 ~ 661
KOO, TAI YOUNG / 2007 / Expression of Recombinant Human Growth Hormone in a Soluble Form inEscherichia coli by Slowing Down the Protein Synthesis Rate / Journal of Microbiolo
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