Isolation of Anammoxosomes From the Aggregate Culture of Ca. Brocadia Sapporoensis and Assembly of Ladderane Liposomes

IF 3.5 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology and Bioengineering Pub Date : 2025-05-08 DOI:10.1002/bit.29011

Tomáš Podzimek, Terezie Cisarová, Michal Dvořák, Barbora Vokatá, Christina Karmann, Jaroslav Hanuš, Martin Balouch, Matěj Malý, Jana Hajšlová, Vojtěch Kouba, Jan Bartáček, František Štěpánek, Petra Lipovová

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引用次数: 0

Abstract

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.

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札幌花楸聚集培养厌氨粘液体的分离及脂质体的组装

厌氧氨氧化菌具有能量高效的氮代谢，它被包裹在由独特的阶梯烷脂组成的厌氧氨酶细胞器中。因此，厌氧氨氧化废物生物质似乎是一个有吸引力的目标，用于分离阶梯烷和随后生产用于药物输送的人工囊泡。本研究提出了一种新的方法，从混合培养的日本札幌Ca. Brocadia sapporoensis聚集体中分离出富含ladderane的厌氨粘液体。与传统的分离方案相比，该方案简化了厌氧氨氧化细胞的预纯化，用蔗糖梯度代替Percoll®，延长了EDTA的应用时间。这种增强和简化的程序有效地去除了EPS和其他碎片，从而产生了对照实验和透射电镜证实的无粘胶体层。本研究首次将所得的脂质体分离物用于制备含和不含纯双棕榈酰磷脂酰胆碱（DPPC）的脂质体。通过TEM和DLS对囊泡进行了表征，并将厌氧氨氧化基阶梯烷掺入囊泡壁中。这些脂质体具有有趣的功能特性，例如在高浓度下增加胶体稳定性，这意味着与仅由DPPC制成的模型脂质体相比，形成聚集体的倾向降低。总的来说，这项研究为将厌氧氨氧化废物转化为药物输送的宝贵资源提供了见解。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology and Bioengineering 工程技术-生物工程与应用微生物

CiteScore

7.90

自引率

5.30%

发文量

280

审稿时长

2.1 months

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