Innovative production of bio-cellulose using a cell-free system derived from a single cell line.

The current study was intended to produce bio-cellulose through a cell-free system developed by disrupting Gluconacetobacter hansenii PJK through bead-beating. Microscopic analysis indicated the complete disruption of cells (2.6 × 10(7) cells/mL) in 20 min that added 95.12 µg/mL protein, 1.63 mM ATP, and 1.11 mM NADH into the medium. A liquid chromatography mass spectrometry/mass spectrometry linear trap quadrupole (LC-MS/MS LTQ) Orbitrap analysis of cell-lysate confirmed the presence of all key enzymes for bio-cellulose synthesis. Under static conditions at 30 °C, microbial and cell-free systems produced 3.78 and 3.72 g/L cellulose, corresponding to 39.62 and 57.68% yield, respectively after 15 days. The improved yield based on consumed glucose indicated the superiority of cell-free system. Based on current findings and literature, we hypothesized a synthetic pathway for bio-cellulose synthesis in the cell-free system. This approach can overcome some limitations of cellulose-producing cells and offers a wider scope for synthesizing cellulose composites with bactericidal elements through in situ synthesizing approaches.

Cellulose production by Gluconacetobacter hansenii in a medium containing ethanol.

The addition of 1% (v/v) ethanol to the basal medium inhibited growth of Gluconacetobacter hansenii but decreased the numbers of non-cellulose producing cells. Cellulose production increased 1.7 times to approx. 2.5 g l(-1) and showed a pattern of mixed growth-associated production. Microbial cells produced rigid pellicle-type bacterial cellulose as the shell of a large lump of bacterial cellulose like a static culture. The inoculum cultivated for 3 d maintained cellulose production by the fifth batch.