Ultrasmall metal nanoclusters (NCs)
are employed in an array of
diagnostic and therapeutic applications due to their tunable photoluminescence,
high biocompatibility, polyvalent effect, ease of modification, and
photothermal stability. However, gold nanoclusters’ (AuNCs’)
intrinsically antimicrobial properties remain poorly explored and
are not well understood. Here, we share an insight into the antimicrobial
action of atomically precise AuNCs based on their ability to passively
translocate across the bacterial membrane. Functionalized by a hydrophilic
modified-bidentate sulfobetaine zwitterionic molecule (AuNC-ZwBuEt)
or a more hydrophobic monodentate-thiolate, mercaptohexanoic acid
(AuNC-MHA) molecule, 2 nm AuNCs were lethal to both Gram-negative Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus bacteria. The bactericidal
efficiency was found to be bacterial strain-, time-, and concentration-dependent.
The direct visualizations of the translocation of AuNCs and AuNC–cell
and subcellular interactions were investigated using cryo-soft X-ray
nano-tomography, transmission electron microscopy (TEM), and scanning
TEM energy-dispersive spectroscopy analyses. AuNC-MHA were identified
in the bacterial cytoplasm within 30 min, without evidence of the
loss of membrane integrity. It is proposed that the bactericidal effect
of AuNCs is attributed to their size, which allows for efficient energy-independent
translocation across the cell membrane. The internalization of both
AuNCs caused massive internal damage to the cells, including collapsed
subcellular structures and altered cell morphology, leading to the
eventual loss of cellular integrity.