Lactobacillus gasseri LA327, isolated from the large intestine tissue in humans, is a bacteriocinogenic strain with two kinds of class IIb bacteriocin structural genes, i.e., those for gassericin T (GT) and acidocin LF221A (Acd LF221A). In this study, DNA sequencing of the genes for GT and Acd LF221A from L. gasseri LA327 revealed that the amino acid sequences for GT corresponded with those for GT genes, except for GatK (histidine kinase). However, Acd LF221A genes had analogues which differed in at least one amino acid residue, to encode a class IIb bacteriocin designated gassericin S (GS). The LA327 strain retained antimicrobial activity after the deletion of the GT structural genes (gatAX); however, both GS and GT activities were lost by deletion of the putative ABC transporter gene (gatT). This indicates that the LA327 strain produces GS and GT and that GS secretion is performed via GT genes with the inclusion of gatT. Homologous expression using deletion mutants of GS and GT, each containing a single peptide, elucidated that GS (GasAX) and GT (GatAX) showed synergistic activity as class IIb bacteriocins and that no synergistic activity was observed between GS and GT peptides. The molecular mass of GS was estimated to be theoretical ca. 5,400 Da by in situ activity assay after SDS-PAGE, clarifying that GS was actually expressed as an active class IIb bacteriocin. Furthermore, the stability of expressed GS to pH, heat, and protease was determined.
IMPORTANCE Bacteriocins are regarded as potential alternatives for antibiotics in the absence of highly resistant bacteria. In particular, two-peptide (class IIb) bacteriocins exhibit the maximum activity through the synergy of two components, and their antimicrobial spectra are known to be relatively wide. However, there are few reports of synergistic activity of class IIb bacteriocins determined by isolation and purification of individual peptides. Our results clarified the interaction of each class IIb component peptide for GT and GS via the construction of homologous mutants, which were not dependent on the purification. These data may contribute to understanding the mechanisms of action by which class IIb bacteriocins exhibit wide antibacterial spectra.
Rapid formation of small mixed-valence luminescent silver clusters via cation-coupled electron-transfer by redox-active polyoxometalate-based porous crystals is reported.
A layered
metal–organic framework (MOF) comprising extra-large
nanographene sheets,
HBC
MOF,
was successfully synthesized using a dicarboxylic acid derivative
of hexa-peri-hexabenzocoronene (HBCLH2), and its structure was characterized by single-crystal X-ray
diffraction analysis. The crystal structure shows that 2D layers composed
of a dinuclear Zn2+ complex unit and HBCL are
located on top of each other through multiple weak interlayer bonds,
affording
HBC
MOF, having three
dimensionally connected nanopores with large nanographene surfaces.
The HBC-based nanographene sheets are anchored to the MOF framework
via two zinc carboxylate linkages and therefore have an axial rotational
freedom. The sorption isotherms of gaseous molecules such as carbon
dioxide and hydrocarbons (acetylene, propane, propylene, benzene,
and cyclohexane) on
HBC
MOF all
displayed a hysteretic profile with reversible structural changes,
as observed by in situ powder X-ray diffraction studies.
Herein, we developed isostructural metal–organic frameworks (MOFs) [Cu1−xPdx(SiF6)(bpy)2] (bpy: 4,4′-bipyridine) (SIFSIX-1-CuPd-3, -5 and -10) containing open metal sites using a partial metal-replacement approach.
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