AVT-Biochemical Engineering, RWTH Aachen University, Aachen, Germany.
iAMB-Institute of Applied Microbiology, RWTH Aachen University, Aachen, Germany.
Biotechnol Bioeng. 2024 Mar;121(3):1076-1089. doi: 10.1002/bit.28633. Epub 2023 Dec 27.
Chemically defined mineral media are widely used in bioprocesses, as these show less batch to batch variation compared with complex media. Nonetheless, the recommended media formulations often lead to the formation of precipitants at elevated pH values. These precipitates are insoluble and reduce the availability of macronutrients to the cells, which can result in limiting growth rates and lower productivity. They can also damage equipment by clogging pipes, hoses, and spargers in stirred tank fermenters. In this study, the observed precipitate was analyzed via X-ray fluorescence spectroscopy and identified as the magnesium ammonium phosphate salt struvite (MgNH PO × 6H O). The solubility of struvite crystals is known to be extremely low, causing the macronutrients magnesium, phosphate, and ammonium to be bound in the struvite crystals. Here, it was shown that struvite precipitates can be redissolved under common fermentation conditions. Furthermore, it was found that the struvite particle size distribution has a significant effect on the dissolution kinetics, which directly affects macronutrient availability. At a certain particle size, struvite crystals rapidly dissolved and provided unlimiting growth conditions. Therefore, struvite formation should be considered during media and bioprocess development, to ensure that the dissolution kinetics of struvite are faster than the growth kinetics.
化学定义的矿物培养基在生物工艺中被广泛使用,因为与复杂培养基相比,这些培养基的批次间变化较小。然而,推荐的培养基配方往往会在较高的 pH 值下形成沉淀物。这些沉淀物是不溶性的,会降低细胞对大量营养素的利用率,从而导致生长速率受限和生产力降低。它们还会通过堵塞搅拌罐发酵罐中的管道、软管和喷射器来损坏设备。在这项研究中,通过 X 射线荧光光谱法对观察到的沉淀物进行了分析,并确定为镁铵磷酸盐盐鸟粪石(MgNH PO × 6H O)。众所周知,鸟粪石晶体的溶解度极低,导致镁、磷酸盐和铵等大量营养素结合在鸟粪石晶体中。这里表明,鸟粪石沉淀物可以在常见的发酵条件下重新溶解。此外,还发现鸟粪石颗粒大小分布对溶解动力学有显著影响,这直接影响大量营养素的可用性。在一定的粒径下,鸟粪石晶体迅速溶解并提供了无限制的生长条件。因此,在开发培养基和生物工艺时应考虑鸟粪石的形成,以确保鸟粪石的溶解动力学比生长动力学更快。