Podzimek Tomáš, Cisarová Terezie, Dvořák Michal, Vokatá Barbora, Karmann Christina, Hanuš Jaroslav, Balouch Martin, Malý Matěj, Hajšlová Jana, Kouba Vojtěch, Bartáček Jan, Štěpánek František, Lipovová Petra
Department of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Prague, Czechia.
Department of Chemical Engineering, University of Chemistry and Technology Prague, Prague, Czechia.
Biotechnol Bioeng. 2025 Aug;122(8):2165-2178. doi: 10.1002/bit.29011. Epub 2025 May 7.
Anammox bacteria wield an energy-efficient nitrogen metabolism enveloped in anammoxosome organelle composed of unique ladderane lipids. Thus, waste anammox biomass seems to be an attractive target for the isolation of ladderanes and subsequent production of artificial vesicles for drug delivery. This study proposed a novel method to isolate ladderane-rich anammoxosomes from aggregate mixed culture of Ca. Brocadia sapporoensis. Compared to conventional isolation protocols, the protocol was simplified by omitting the prepurification of anammox cells, replacing Percoll® with a sucrose gradient and prolonging the application of EDTA. This enhanced and simplified procedure efficiently removed EPS and other debris, thus yielding the layer of anammoxosomes as confirmed by control experiments and TEM. For the first time, the resulting ladderane isolates were used for the preparation of liposomes, both with and without the addition of pure dipalmitoylphosphatidylcholine (DPPC). Vesicles were successfully created, characterised by TEM and DLS, and anammox-based ladderanes were incorporated into their walls. These liposomes had interesting functional properties such as increased colloid stability at elevated concentrations, meaning a reduced tendency to form aggregates compared to model liposomes made solely of DPPC. Overall, this study offers insights into converting waste anammox biomass into a valuable resource for drug delivery.
厌氧氨氧化细菌拥有一种高效的氮代谢方式，其被包裹在由独特的梯形烷脂质构成的厌氧氨氧化体细胞器中。因此，废弃的厌氧氨氧化生物质似乎是分离梯形烷以及随后生产用于药物递送的人工囊泡的一个有吸引力的目标。本研究提出了一种从嗜盐碱布罗卡德氏菌（Candidatus Brocadia sapporoensis）的聚集体混合培养物中分离富含梯形烷的厌氧氨氧化体的新方法。与传统的分离方案相比，该方案通过省略厌氧氨氧化细胞的预纯化、用蔗糖梯度代替Percoll® 以及延长EDTA的使用时间而得到简化。这种改进和简化的程序有效地去除了胞外聚合物和其他碎片，从而得到了厌氧氨氧化体层，对照实验和透射电子显微镜（TEM）证实了这一点。所得的梯形烷分离物首次被用于制备脂质体，制备过程中添加和不添加纯二棕榈酰磷脂酰胆碱（DPPC）。成功制备出了囊泡，通过TEM和动态光散射（DLS）对其进行了表征，并且基于厌氧氨氧化的梯形烷被整合到了它们的壁中。这些脂质体具有有趣的功能特性，例如在较高浓度下具有更高的胶体稳定性，这意味着与仅由DPPC制成的模型脂质体相比，其形成聚集体的倾向降低。总体而言，本研究为将废弃的厌氧氨氧化生物质转化为用于药物递送的宝贵资源提供了见解。
