Cell wall mechanical properties as measured with bacterial thread made from Bacillus subtilis

Abstract

Engineering approaches used in the study of textile fibers have been applied to the measurement of mechanical properties of bacterial cell walls by using the Bacillus subtilis bacterial thread system. Improved methods have been developed for the production of thread and for measuring its mechanical properties. The best specimens of thread produced from cultures of strain FJ7 grown in TB medium at 20 degrees C varied in diameter by a factor of 1.09 over a 30-mm thread length. The stress-strain behavior of cell walls was determined over the range of relative humidities between 11 and 98%. Measurements of over 125 specimens indicated that cell wall behaved like other viscoelastic polymers, both natural and man-made, exhibiting relaxation under constant elongation and recovery upon load removal. This kinetic behavior and also the cell wall strength depended greatly on humidity. The recovery from extension observed after loading even up to a substantial fraction of the breaking load indicated that the properties measured were those of cell wall material rather than of behavior of the thread assemblage. Control experiments showed that neither drying of thread nor the length of time it remained dry before testing influenced the mechanical properties of the cell walls. Specimens drawn from TB medium and then washed in water and redrawn were found to be stiffer and stronger than controls not washed. However, tensile properties were not changed by exposure of cells to lysozyme before thread production. This suggests that glycan backbones are not arranged along the length of the cell cylinder. The strength of the cell wall in vivo was estimated by extrapolation to 100% relative humidity to be about 3 N/mm2. Walls of this strength would be able to bear a turgor pressure of 6 atm (ca. 607.8 kPa), but if the increase in strength of water-washed threads was appropriate, the figure could be 24 atm (ca. 2,431.2 kPa).