Abstract Bacterial cellulose (BC) has gained significant interest for various applications due to its useful properties which include chemical stability and strong physical properties as well as its biocompatibility. However, conventional fabrication methods are limited by a lack of control over the form, particularly depth and thickness, restricting BC applications to 2D planar shapes. The production of BC is constrained by the formation of a pellicle at the liquid–air interface. To overcome this constraint and enable continuous pellicle formation, a method was established to combine intermittent batch feeding with a supportive mesh scaffold. Intermittent feeding supplies nutrients to the pellicle, promoting sustained formation at the liquid–air interface, while the supporting mesh anchors the initial pellicle to prevent submersion. This approach facilitates the continuous growth of BC pellicle at a controlled rate. Moreover, the method described here results in a single BC pellicle with enhanced thickness, weight (both wet and dry states), water-holding capacity and mechanical strength. Continuous growth is achievable with ongoing nutrient supply, allowing for precise control over the final pellicle thickness. This culturing method is scalable and has been tested for volumes ranging from 250 mL to 10 L, and significantly increases glucose-to-cellulose conversion (3.4-fold) and water utilization efficiency (2.4-fold).
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