site > International journal > (2008) Effects of β-mercaptoethanol and hydrogen peroxide on enzymatic conversion of human proinsulin into insulin.

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작성일 : 12-07-17 17:56

(2008) Effects of β-mercaptoethanol and hydrogen peroxide on enzymatic conversion of human proinsulin into insulin.

글쓴이 : 박용철

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http://www.ncbi.nlm.nih.gov/pubmed/18633302 [991]

http://www.jmb.or.kr/journal/viewJournal.html?year=2008&vol=18&num=5&p… [383]

Effects of β-mercaptoethanol and hydrogen peroxide on enzymatic conversion of human proinsulin into insulin.

Young-Jin Son, Chang-Kyu Kim, Byoung-Taek Choi, Yong-Cheol Park and Jin-Ho Seo*
Journal of Microbiology and Biotechnology 18(5): 983-989. 2008.05.28.
(SCIE I/F: 1.381)

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 beta-mercaptoethanol. To make authentic insulin, recombinant preproinsulin is modified enzymatically by trypsin and carboxypeptidase B. The effects of beta-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 beta-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 beta-mercaptoethanol and thereby reduced the negative effects of beta-mercaptoethanol considerably. Elimination of the impurity 3 and other derivatives by the addition of over 10 mM hydrogen peroxide in the presence of beta-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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