The bacterium
Actinoplanes missouriensis
belongs to the genus
Actinoplanes
, a prolific source of useful natural
products. This microbe forms
globular structures called sporangia, which contain many dormant spores.
Recent studies using transmission electron microscopy have shown that
the
A. missouriensis
sporangium membrane
has an unprecedented three-layer structure, but its molecular components
remain unclear. Here, we present multimodal (spontaneous Raman scattering,
coherent anti-Stokes Raman scattering, second harmonic generation,
sum frequency generation, and third-order sum frequency generation)
label-free molecular imaging of intact
A. missouriensis
sporangia. Spontaneous Raman imaging assisted with multivariate
curve resolution–alternating least-squares analysis revealed
a novel component in the sporangium membrane that exhibits unique
Raman bands at 1550 and 1615 cm
–1
in addition to
those characteristic of lipids. A plausible candidate for this component
is an unsaturated carbonyl compound with an aliphatic moiety derived
from fatty acid. Furthermore, second harmonic generation imaging revealed
that a layer(s) of the sporangium membrane containing this unknown
component has an ordered, noncentrosymmetric structure like fibrillar
proteins and amylopectin. Our results suggest that the sporangium
membrane is a new type of biological membrane, not only in terms of
architecture but also in terms of components. We demonstrate that
multimodal molecular imaging with Raman scattering as the core technology
will provide a promising platform for interrogating the chemical components,
whether known or unknown, of diverse biological structures produced
by microbes.