Tofteng A Pernille, Jensen Knud J, Schäffer Lauge, Hoeg-Jensen Thomas
Faculty of Life Sciences, University of Copenhagen, Frederiksberg, Denmark.
Chembiochem. 2008 Dec 15;9(18):2989-96. doi: 10.1002/cbic.200800430.
Insulin is a peptide hormone consisting of 51 amino acids in two chains with three disulfide bridges. Human insulin and various analogues are used for the treatment of diabetes and are produced recombinantly at ton scale. Herein, we report the chemical synthesis of insulin by the step-wise, Fmoc-based, solid-phase synthesis of single-chain precursors with solubilising extensions, which under redox conditions, spontaneously fold with the correct pairing of the three disulfide bridges. The folded, single-chain, insulin precursors can be transformed into bioactive two-chain desB30 insulin by the simultaneous removal of the solubilising extension (4-5 residues) and the chain-bridging C-peptide (3-5 residues) by employing Achromobacter lyticus protease--a process well-known from the yeast-based recombinant production of insulin. The overall yields of synthetic insulins were as much as 6 %, and the synthetic process was straightforward and not labour intensive.
胰岛素是一种肽激素，由两条链中的51个氨基酸组成，有三个二硫键。人胰岛素和各种类似物用于治疗糖尿病，并且以吨级规模通过重组生产。在此，我们报告了通过基于Fmoc的逐步固相合成单链前体并带有增溶延伸部分来化学合成胰岛素，该单链前体在氧化还原条件下，通过三个二硫键的正确配对自发折叠。通过使用溶杆菌蛋白酶同时去除增溶延伸部分（4 - 5个残基）和链桥接C肽（3 - 5个残基），折叠后的单链胰岛素前体可以转化为具有生物活性的双链去B30胰岛素——这是基于酵母的胰岛素重组生产中熟知的过程。合成胰岛素的总产率高达6%，并且合成过程简单，不耗费大量人力。
