BACKGROUND Insulin controls hyperglycemia caused by diabetes, and virtually all treatments require exogenous insulin. However, the product's extensive post-translational modifications have hindered the manufacture of recombinant insulin. RESULT Here we report a novel production method for a monomeric B22Asp desB30 insulin analog (B22D desB30 insulin). Its precursor, DPIP, is fused to an N-terminal chitin-binding domain and intein self-cleavage tag. The fusion protein is expressed and purified from E. coli and immobilized on chitin resins. DPIP is then released using an optimized pH shift and converted to mature insulin via trypsin digest. The resulting product appears monomeric, > 90% pure and devoid of any exogenous enzyme. CONCLUSION Thus, biologically active insulin analog can be efficiently produced in bacteria and potentially applicable in the treatment of human diabetes.

中文翻译：

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内含蛋白介导的单体B22Asp desB30胰岛素的重组表达。

背景技术胰岛素控制由糖尿病引起的高血糖症，并且实际上所有治疗都需要外源性胰岛素。但是，该产品广泛的翻译后修饰阻碍了重组胰岛素的生产。结果在这里我们报告了一种新的生产方法，用于单体B22Asp desB30胰岛素类似物（B22D desB30胰岛素）。它的前体DPIP与N端几丁质结合域和内含肽自我切割标签融合。从大肠杆菌表达和纯化融合蛋白，并将其固定在几丁质树脂上。然后使用优化的pH转换释放DPIP，并通过胰蛋白酶消化将其转化为成熟的胰岛素。所得产物为单体，纯度＞ 90％，且不含任何外源酶。结论因此，