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JoVE Encyclopedia of Experiments
Microbiology
纤维素生产细菌的细菌纳米纤维素薄膜制备
纤维素生产细菌的细菌纳米纤维素薄膜制备
Encyclopedia of Experiments
Microbiology
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Encyclopedia of Experiments Microbiology
Preparation of Bacterial Nanocellulose Pellicles from Cellulose-Producing Bacteria

纤维素生产细菌的细菌纳米纤维素薄膜制备

Protocol
304 Views
02:52 min
November 10, 2025
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Please note that some of the translations on this page are AI generated. Click here for the English version.

Transcript

取一个装有含甘露醇营养介质的微型板。

在每个井中接种产纤维素的细菌,并在静止条件下培养。

在培养过程中,细菌吸收甘露醇并将其代谢为葡萄糖。

这些葡萄糖被运输到嵌入细菌膜中的纤维素合成酶复合物,并聚合成葡聚糖链。

这些链向细胞外空间伸出,结晶成微纤维。

微纤维束成带状结构,形成三维纳米纤维素基质。

随着纳米纤维素的产生增加,它在空气-液体界面积累,逐渐增厚成胶状薄膜,形成井形模塑。

采集菌膜,并将其培养在碱性溶液中,以裂解细菌细胞,然后去除残留培养基。

用水反复冲洗膜膜,以消除残留的碱性溶液。将胶囊浸泡在水中以防止脱水。然后进行蒸汽消毒,以确保后续应用时保持无菌状态。

将2毫升液体培养基转移到24孔组织培养板的每个孔中。从已接种培养皿中取两个植落,接种针头同步,放入组织培养皿的第一个孔中。对剩余的23口井重复同样的作。

将组织培养板在30摄氏度下孵育七天。这将产生共24个直径16毫米、厚度约2至3毫米的BNC膜。从培养基中采集BNC膜,并用200毫升1%氢氧化钠溶液、50摄氏度消毒一小时,以去除所有 木质菌的痕迹。

丢弃溶液,加入200毫升新鲜制备的1%氢氧化钠溶液。重复同一过程,直到溶液中的BNC薄膜呈现半透明。用水冲洗BNC胶膜三次,然后在室温高纯水中保存。

确保BNC薄膜完全浸没在水中,且任何时候都不能干燥。然后,将BNC薄膜以121摄氏度高压灭菌20分钟。

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