Enzyme digestion conversion method of recombinant human insulin precursor

Abstract

The invention discloses an enzyme digestion conversion method of a recombinant human insulin precursor. The enzyme digestion conversion method comprises the following steps: (1) adding a primarily purified recombinant human insulin precursor solution into a buffer solution, and adjusting a concentration and a pH value to obtain an enzyme digestion reaction substrate mixed solution; and (2) adding recombinant lysyl endonuclease into the enzyme digestion reaction substrate mixed solution to carry out enzyme digestion conversion reaction on the recombinant human insulin precursor. The recombinant lysyl endonuclease is selected as a tool enzyme, and compared with common recombinant trypsin, the recombinant lysyl peptide endonuclease has advantages that less enzyme is needed by unit precursor protein, enzyme digestion site is single, enzyme digestion process is more efficient, and enzyme digestion conversion efficiency of a human insulin precursor reaches 95% or above; when enzyme digestion is amplified, worry about excessive enzyme digestion is avoided, and purity of target protein obtained after enzyme digestion is higher; the optimum pH range of enzyme digestion of the recombinant lysyl endonuclease is farther from pI of the protein before and after enzyme digestion, so that stability of the target protein is facilitated, and tolerance range to a salt concentration in the enzyme digestion environment is wider.

Description

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AI Technical Summary

Benefits of technology

This patented technology helps improve the ability of humans's own beta cells (β cell) to break down their proteins into amino acids that are needed for growth or other functions like muscle contractions when they die due to lacking enough calories during exercise. By selecting specific types of cleavage enzymatic tools with fewer parts relative to traditional methods, this new approach simplifies the manufacturing processes while maintaining high levels of accuracy. Additionally, these techniques reduce complexity and simplify processing. Overall, this innovation provides better ways to produce biologically active agents such as hormones through genetic engineering rather than chemistry.

Problems solved by technology

Technological Problem: Current methods for producing high levels of glucose regulate blood sugar level through controlling various metabolites such as glycosaminoglutathione (GSH) sulfate and other substances like uricase activity during fasting periods. However, these techniques may also affect certain proteins involved in signal transductions between cells and tissues, leading to decreased accuracy when analyzed over time.

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