Rupcic Zeljka, Hüttel Stephan, Bernecker Steffen, Kanaki Sae, Stadler Marc
Department Microbial Drugs, Helmholtz Centre for Infection Research GmbH, Inhoffenstraße 7, 38124 Braunschweig, Germany.
German Centre for Infection Research (DZIF), partner site Hannover-Braunschweig, 38124 Braunschweig, Germany.
Bioengineering (Basel). 2018 Jun 5;5(2):42. doi: 10.3390/bioengineering5020042.
A method was established for the production of 1.2-fold and 4.2-fold increased amounts of the antiviral and central nervous system-active lantipeptides, labyrinthopeptins A1 and A2, respectively, isolated from the actinobacterium , to enable production in gram scale. We then performed in vivo characterization of this promising compound class. The labyrinthopeptins A1 and A2 have similar chemical structures and physical properties but differ drastically in their bioactivities. Therefore, large quantities of highly pure material are required for pharmacological studies. An effective methodology was established for the first time for their production in bioreactors, their separation involving gel permeation chromatography on LH20 material, followed by reversed phase-high performance liquid chromatography. With an optimized methodology, 580 mg of labyrinthopeptin A1 and 510 mg of labyrinthopeptin A2 were quantitatively isolated with recovery rates of 72.5% and 42.3% from 7.5 L of culture broth, respectively. However, the fermentation that had already resulted in maximum yields of over 100 mg/L of both target molecules after 300 h in a 10-L scale bioreactor, still requires further optimisation.
建立了一种方法，用于分别从放线菌中生产抗病毒和具有中枢神经系统活性的羊毛硫肽（迷宫肽A1和A2），产量分别提高1.2倍和4.2倍，以实现克级规模的生产。然后，我们对这一有前景的化合物类别进行了体内表征。迷宫肽A1和A2具有相似的化学结构和物理性质，但生物活性差异很大。因此，药理学研究需要大量高纯度的材料。首次建立了一种有效的方法，用于在生物反应器中生产它们，并通过在LH20材料上进行凝胶渗透色谱，随后进行反相高效液相色谱进行分离。采用优化的方法，从7.5升培养液中分别定量分离出580毫克迷宫肽A1和510毫克迷宫肽A2，回收率分别为72.5%和42.3%。然而，在10升规模的生物反应器中，经过300小时发酵后，两种目标分子的产量已达到最高，超过100毫克/升，但仍需要进一步优化。
