The invention discloses a preparation method of recombinant
human insulin, which relates to the
protein polypeptide type
medicine field. The technical problem needed to be solved by the invention aims at providing a preparation method for
Escherichia coli to express the recombinant
human insulin in order to overcome the technical disadvantages in the prior art of using the poisonous and harmful substance CNBr and having a complex production process. The preparation method for the recombinant
human insulin of the invention comprises the steps of:
thallus crashing, inclusion body dissolving,
proinsulin sulfonating, purifying
proinsulin, and
proinsulin renaturation,
enzyme cutting transmission, etc., and the preparation method is characterized in that during the inclusion body dissolving process, the -SH of proinsulin is transformed into -SSO3<->, and then human
insulin is obtained through purifying the sulfonated proinsulin, reduction renaturation and
enzyme cutting transmission by positive
ion, and then through the negative
ion exchange layer purification. The preparation method of the recombinant human
insulin of the invention does not use the CNBr and has simple technique with high human
insulin yield coefficient and purity.