Abstract

The mechanical properties of Rhodococcus RC291 were measured using force spectroscopy equipped with a bacterial cell probe. Rhodococcal cells in the late growth stage of development were found to have greater adhesion to a silicon oxide surface than those in the early growth stage. This is because there are more extracellular polymeric substances (EPS) that contain nonspecific binding sites available on the cells of late growth stage. It is found that EPS in the late exponential phase are less densely bound but consist of chains able to extend further into their local environment, while the denser EPS at the late stationary phase act more to sheath the cell. Contraction and extension of the EPS could change the density of the binding sites, and therefore affect the magnitude of the adhesion force between the EPS and the silicon oxide surface. By treating rhodococcal EPS as a surface-grafted polyelectrolyte layer and using scaling theory, the interaction between EPS and a solid substrate was modelled for the cell approaching the surface which revealed that EPS possess a large capacity to store charge. Changing the pH of the surrounding medium acts to change the conformation of EPS chains.

Highlights

  • Oil spills and toxic compounds discharged from industrial activities and agriculture are examples of processes causing significant hydrocarbon contamination

  • extracellular polymeric substances (EPS) of the stationary phase are capable of storing more charge than EPS of the exponential phase in the corresponding electric double layer

  • During the retraction of the cell probe from the model silicon oxide surface, more adhesive interactions were found between rhodococcal cells of the stationary phase and the substrate, which were attributed to the chemical composition of the EPS

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Summary

Introduction

Oil spills and toxic compounds discharged from industrial activities and agriculture are examples of processes causing significant hydrocarbon contamination. It is reasonable to treat EPS as weak polyelectrolytes, i.e. long chain macromolecules possessing ionizable groups [27] This allows quantitative modelling of the repulsive force between the EPS and a solid surface when they approach each other, taking into account both electrostatic and steric forces [28]. Several studies have been carried out to investigate steric interactions, adhesion, and viscoelasticity of the EPS of different bacterial strains [24, 36,37,38,39,40], but few have examined the contribution of EPS to the mechanical properties of whole cell. By attaching cells from different growth stages to the AFM cantilever, the mechanical properties of Rhodococcus are studied in terms of their: (1) charge storage, (2) water retention capability, and (3) adhesion to a model surface

Cultivation of Rhodococcus
Preparation of bacterial cell probe
Force spectroscopy
Contact angle measurements
Preparation of model substrates
Rhodococcal cell surface properties
Mechanical properties of rhodococcal cell under compression
Electrosteric forces between rhodococcal cell and a silicon oxide substrate
Adhesion of rhodococcal cell to a silicon oxide surface
General discussion
Conclusion
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