JoVE Encyclopedia of Experiments
Microbiology
0 views • 4:20 min • September 26th, 2025
Begin with a multi-well plate containing growth media pre-inoculated with cellulose-producing bacteria.
In the treated well, the media is supplemented with 2-chloroethylphosphonic acid (CEPA).
Seal the plate to prevent gas exchange and incubate.
During incubation, CEPA hydrolyzes and releases ethylene, a gaseous molecule.
Ethylene activates bacterial cellulose synthase, which utilizes cytosolic UDP-glucose to synthesize cellulose.
Over time, the bacteria extrude cellulose through their membrane.
This promotes bacterial self-assembly at the air-liquid interface, forming a floating cellulose-based biofilm, or pellicle.
Following incubation, transfer the pellicles to a fresh multi-well plate. Add an alkaline solution and incubate at a higher temperature to lyse the cells.
Remove the alkaline solution. Repeatedly wash with agitation to remove cellular debris while retaining the cellulose.
Dry and weigh the pellicles. A higher dry weight in the CEPA-treated samples compared to the control indicates enhanced bacterial cellulose production in response to ethylene.
The pellicle assay is a standard analysis tool for bacterial cellulose production. PH and morphological analysis are detailed in text pr
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