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The type 9 secretion system (T9SS) is the protein export pathway of bacteria of the Gram-negative Fibrobacteres–Chlorobi–Bacteroidetes superphylum and is an essential determinant of pathogenicity in severe periodontal disease. The central element of the T9SS is a so-far uncharacterized protein-conducting translocon located in the bacterial outer membrane. Here, using cryo-electron microscopy, we provide structural evidence that the translocon is the T9SS protein SprA. SprA forms an extremely large (36-strand) single polypeptide transmembrane β-barrel. The barrel pore is capped on the extracellular end, but has a lateral opening to the external membrane surface. Structures of SprA bound to different components of the T9SS show that partner proteins control access to the lateral opening and to the periplasmic end of the pore. Our results identify a protein transporter with a distinctive architecture that uses an alternating access mechanism in which the two ends of the protein-conducting channel are open at different times. Cryo-electron microscopy structures of the protein-conducting translocon of the type 9 secretion system reveal its architecture and mechanism of translocation.

Polysaccharides are recognized for their extensive biological functions, holding significant promise for applications in both medicine and food industries. However, their utilization is frequently constrained by challenges such as high molecular weights and indistinct sugar chain structures. Recently, two novel bacterial strains, N6 T and J3 T , were isolated from the Nakdong River in Korea. These strains, which belong to the phylum Bacteroidota , are Gram-stain-negative, non-motile, aerobic, rod-shaped bacteria and have shown polysaccharide-degrading capabilities. Through comprehensive analyses, including 16S rRNA gene sequencing, whole-genome sequencing, and detailed morphological, physiological, and chemotaxonomic characterizations, these strains have been identified as new species within the genus Flavobacterium . KEGG pathway analysis further confirmed their robust capabilities for carbohydrate utilization. Additional investigations using the dbCAN and dbCAN-PUL databases identified the presence of carbohydrate-hydrolyzing enzymes (CAZymes) and polysaccharide utilization loci (PULs) within these strains, suggesting their potential to degrade various polysaccharides. Subsequent in vitro growth experiments demonstrated that strains N6 T and J3 T can degrade chitin, β-glucan, κ-carrageenan, and cellulose. Given their diverse polysaccharide degradation abilities, these strains are formally proposed to be named Flavobacterium polysaccharolyticum sp. nov. and Flavobacterium aureirubrum sp. nov. The type strains are designated as N6 T (= KCTC 102173 T  = GDMCC 1.4609 T ) and J3 T (= KCTC 102172 T  = GDMCC 1.4608 T ), respectively.

Many plants associate with endophytic microbes that improve root phosphorus (P) uptake. Understanding the interactions between roots and endophytes can enable efforts to improve P utilization. Here, we characterize the interactions between lateral roots of endophytes in a core collection of 50 rapeseed ( Brassica napus L.) genotypes with differing sensitivities to low P conditions. With the correlation analysis result between bacterial abundance and plant physiological indices of rapeseeds, and inoculation experiments on plates and soil, we identify one Flavobacterium strain (C2) that significantly alleviates the P deficiency phenotype of rapeseeds. The underlying mechanisms are explored by performing the weighted gene coexpression network analysis (WGCNA), and conducting genome-wide association studies (GWAS) using Flavobacterium abundance as a quantitative trait. Under P-limited conditions, C2 regulates fatty acid and lipid metabolic pathways. For example, C2 improves metabolism of linoleic acid, which mediates root suberin biosynthesis, and enhances P uptake efficiency. In addition, C2 suppresses root jasmonic acid biosynthesis, which depends on α -linolenic acid metabolism, improving C2 colonization and activating P uptake. This study demonstrates that adjusting the endophyte composition can modulate P uptake in B. napus plants, providing a basis for developing agricultural microbial agents. Based on the correlation analysis of various omics data from Brassica napus L., authors identify one Flavobacterium strain C2 which mitigates P deficiency by regulating fatty acid metabolism and enhancing P uptake, indicating potential strategies to improve P utilization in rapeseeds.

The regeneration of bioavailable phosphate from immobilized organophosphorus represents a key process in the global phosphorus cycle and is facilitated by enzymes known as phosphatases. Most bacteria possess at least one of three phosphatases with broad substrate specificity, known as PhoA, PhoX, and PhoD, whose activity is optimal under alkaline conditions. The production and activity of these phosphatases is repressed by phosphate availability. Therefore, they are only fully functional when bacteria experience phosphorus-limiting growth conditions. Here, we reveal a previously overlooked phosphate-insensitive phosphatase, PafA, prevalent in Bacteroidetes, which is highly abundant in nature and represents a major route for the regeneration of environmental phosphate. Using the enzyme from Flavobacterium johnsoniae, we show that PafA is highly active toward phosphomonoesters, is fully functional in the presence of excess phosphate, and is essential for growth on phosphorylated carbohydrates as a sole carbon source. These distinct properties of PafA may expand the metabolic niche of Bacteroidetes by enabling the utilization of abundant organophosphorus substrates as C and P sources, providing a competitive advantage when inhabiting zones of high microbial activity and nutrient demand. PafA, which is constitutively synthesized by soil and marine flavobacteria, rapidly remineralizes phosphomonoesters releasing bioavailable phosphate that can be acquired by neighboring cells. The pafA gene is highly diverse in plant rhizospheres and is abundant in the global ocean, where it is expressed independently of phosphate availability. PafA therefore represents an important enzyme in the context of global biogeochemical cycling and has potential applications in sustainable agriculture.

Colony spreading of Flavobacterium johnsoniae is shown to include gliding motility using the cell surface adhesin SprB, and is drastically affected by agar and glucose concentrations. Wild-type (WT) and Δ sprB mutant cells formed nonspreading colonies on soft agar, but spreading dendritic colonies on soft agar containing glucose. In the presence of glucose, an initial cell growth-dependent phase was followed by a secondary SprB-independent, gliding motility-dependent phase. The branching pattern of a Δ sprB colony was less complex than the pattern formed by the WT. Mesoscopic and microstructural information was obtained by atmospheric scanning electron microscopy (ASEM) and transmission EM, respectively. In the growth-dependent phase of WT colonies, dendritic tips spread rapidly by the movement of individual cells. In the following SprB-independent phase, leading tips were extended outwards by the movement of dynamic windmill-like rolling centers, and the lipoproteins were expressed more abundantly. Dark spots in WT cells during the growth-dependent spreading phase were not observed in the SprB-independent phase. Various mutations showed that the lipoproteins and the motility machinery were necessary for SprB-independent spreading. Overall, SprB-independent colony spreading is influenced by the lipoproteins, some of which are involved in the gliding machinery, and medium conditions, which together determine the nutrient-seeking behavior.

This study presents taxonomic description of two novel diesel-degrading, psychrophilic strains: Kopri-42 T and Kopri-43, isolated during screening of oil-degrading psychrotrophs from oil-contaminated Arctic soil. A preliminary 16S rRNA gene sequence and phylogenetic tree analysis indicated that these Arctic strains belonged to the genus Flavobacterium , with the nearest relative being Flavobacterium psychrolimnae LMG 22018 T (98.9% sequence similarity). The pairwise 16S rRNA gene sequence identity between strains Kopri-42 T and Kopri-43 was 99.7%. The DNA-DNA hybridization value between strain Kopri-42 T and Kopri-43 was 88.6 ± 2.1% indicating that Kopri-42 T and Kopri-43 represents two strains of the same genomospecies. The average nucleotide identity and in silico DNA-DNA hybridization values between strain Kopri-42 T and nearest relative F . psychrolimnae LMG 22018 T were 92.4% and 47.9%, respectively. These values support the authenticity of the novel species and confirmed the strain Kopri-42 T belonged to the genus Flavobacterium as a new member. The morphological, physiological, biochemical and chemotaxonomic data also distinguished strain Kopri-42 T from its closest phylogenetic neighbors. Based on the polyphasic data, strains Kopri-42 T and Kopri-43 represents a single novel species of the genus Flavobacterium , for which the name Flavobacterium petrolei sp. nov. is proposed. The type strain is Kopri-42 T (=KEMB 9005-710 T  = KACC 19625 T  = NBRC 113374 T ).

Light-driven proton-pumping rhodopsins are widely distributed in many microorganisms. They convert sunlight energy into proton gradients that serve as energy source of the cell. Here we report a new functional class of a microbial rhodopsin, a light-driven sodium ion pump. We discover that the marine flavobacterium Krokinobacter eikastus possesses two rhodopsins, the first, KR1, being a prototypical proton pump, while the second, KR2, pumps sodium ions outward. Rhodopsin KR2 can also pump lithium ions, but converts to a proton pump when presented with potassium chloride or salts of larger cations. These data indicate that KR2 is a compatible sodium ion–proton pump, and spectroscopic analysis showed it binds sodium ions in its extracellular domain. These findings suggest that light-driven sodium pumps may be as important in situ as their proton-pumping counterparts. Light-driven proton-pumping rhodopsins are widely distributed in microorganisms and convert sunlight energy into proton gradients. Inoue et al . report the discovery of a light-driven sodium ion pump from marine bacteria.

Naturally occurring photonic structures are responsible for the bright and vivid coloration in a large variety of living organisms. Despite efforts to understand their biological functions, development, and complex optical response, little is known of the underlying genes involved in the development of these nanostructures in any domain of life. Here, we used Flavobacterium colonies as a model system to demonstrate that genes responsible for gliding motility, cell shape, the stringent response, and tRNA modification contribute to the optical appearance of the colony. By structural and optical analysis, we obtained a detailed correlation of how genetic modifications alter structural color in bacterial colonies. Understanding of genotype and phenotype relations in this system opens the way to genetic engineering of on-demand living optical materials, for use as paints and living sensors.

The type 9 secretion system (T9SS) is fundamental to bacterial gliding motility, pathogenesis, and surface colonization. Our findings reveal that the C-terminal region of the conveyor belt-associated protein GldJ functions as a molecular switch which is capable of reversing the rotational direction of T9SS. Through the coordinated actions of the T9SS stator units (akin to a driving motor), the GldK ring (the gear that converts rotational energy into linear movement), and GldJ, this machinery forms a smart conveyor belt system reminiscent of flexible or cognitive mechanical conveyors. Such advanced conveyors can alter their direction to adapt to shifting demands. Here, we show that the bacterial T9SS similarly adjusts its rotational bias based on feedback from the conveyor belt-associated protein GldJ. This dual-role feedback mechanism underscores an evolved, controllable biological snowmobile, offering new avenues for studying how bacteria fine-tune motility in dynamic environments.

Background During the screening of pigment-producing microbes from domestic sources, 102 yellow- or orange-pigmented bacteria were isolated. Among these, two novel Flavobacterium strains, F. sedimentum SUN046 T and F. fluvius SUN052 T , were identified as zeaxanthin producers. A polyphasic taxonomic characterization, combined with comparative genomic analysis of 45 Flavobacterium species, was conducted to determine their taxonomic positions and explore potential evolutionary relationships in zeaxanthin biosynthesis. Results Both strains utilized the mevalonic acid (MVA) pathway and possessed the crt gene cluster ( crtB , crtI , crtY / crtY cd , and crtZ ). Strain SUN046 T exhibited unique features in the carotenoid biosynthesis pathway, notably the absence of HMG-CoA synthase (HMGCS) in the upper MVA pathway and the presence of the rare lycopene β -cyclase crtY cd , which is uncommon among bacteria. The CrtY cd in SUN046 T possessed a single active site and direct lycopene-binding modes. Conversely, CrtY in SUN052 T exhibited multiple active sites, which is flavin adenine dinucleotide (FAD) dependent. These structural differences has impacted catalytic efficiencies, as evidenced by zeaxanthin yields of 6.49 µg/mL in SUN046 T and 13.23 µg/mL in SUN052 T . Variations in carotenoid biosynthetic pathway among other Flavobacterium species were also observed. Conclusion These findings suggest that both strains represent valuable new resources for zeaxanthin production and provide foundational insights for biotechnological applications involving the genus Flavobacterium , highlighting the genetic and evolutionary complexity of microbial carotenoid biosynthesis.