Molecular and Chemical Analysis of the Lipopolysaccharide from Aeromonas hydrophila Strain AH-1 (Serotype O11)

Merino, Susana; Canals, Rocı́o; Knirel, Yuriy A.; Tomás, Juan M.

doi:10.3390/md13042233

Cited by 18 publications

(18 citation statements)

References 41 publications

(46 reference statements)

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“…7), some conserved functional properties clearly emerge. The carbohydrate backbones of the OPS from K. pneumoniae O12 and Aeromonas hydrophila AH-1 are identical, although AH-1 OPS also possesses variable acetylation at the 2 and 3 positions of the Rhap residue (38). The OPS-biosynthesis genetic locus of AH-1 contains a putative acetyltransferase gene.…”

Section: Discussionmentioning

confidence: 96%

Single polysaccharide assembly protein that integrates polymerization, termination, and chain-length quality control

Williams

Ovchinnikova

Koizumi

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Lipopolysaccharides (LPS) are essential outer membrane glycolipids in most gram-negative bacteria. Biosynthesis of the O-antigenic polysaccharide (OPS) component of LPS follows one of three widely distributed strategies, and similar processes are used to assemble other bacterial surface glycoconjugates. This study focuses on the ATP-binding cassette (ABC) transporter-dependent pathway, where glycans are completed on undecaprenyl diphosphate carriers at the cytosol:membrane interface, before export by the ABC transporter. We describe Raoultella terrigena WbbB, a prototype for a family of proteins that, remarkably, integrates several key activities in polysaccharide biosynthesis into a single polypeptide. WbbB contains three glycosyltransferase (GT) modules. Each of the GT102 and GT103 modules characterized here represents a previously unrecognized GT family. They form a polymerase, generating a polysaccharide of [4)-α-Rhap-(1→3)-β-GlcpNAc-(1→] repeat units. The polymer chain is terminated by a β-linked Kdo (3-deoxy-D-manno-oct-2-ulosonic acid) residue added by a third GT module belonging to the recently discovered GT99 family. The polymerase GT modules are separated from the GT99 chain terminator by a coiled-coil structure that forms a molecular ruler to determine product length. Different GT modules in the polymerase domains of other family members produce diversified OPS structures. These findings offer insight into glycan assembly mechanisms and the generation of antigenic diversity as well as potential tools for glycoengineering.microbial glycobiology | lipopolysaccharides | glycosyltransferases | glycan biosynthesis | molecular ruler B acterial surfaces possess an array of complex glycoconjugates (sugar-containing macromolecules) that play varied and vital roles in the biology of these organisms. In pathogens, glycoconjugates participate in adhesion, biofilm formation, and interaction with innate and adaptive immune responses. For example, in gram-negative bacteria such as Escherichia coli, long glycan chains in capsular polysaccharides and the O-antigen polysaccharide (OPS) components of lipopolysaccharide (LPS) molecules typically confer resistance to opsonophagocytosis and complement-mediated killing (1, 2). Effective protection depends on the amount and surface distribution of glycan, as well as glycan chain lengths suited for a given purpose. As an example, shorter OPS chains offer no resistance to complementmediated killing, whereas chains longer than an optimal size potentially represent an unnecessary energy cost while offering no additional advantage (3, 4). Despite its fundamental importance in colonization and virulence of bacterial pathogens, the molecular mechanisms of glycan chain-length regulation are often poorly understood (5). However, OPS biosynthesis provides influential prototypes for understanding the guiding principles underpinning chainlength regulatory mechanisms in bacterial glycoconjugates in general.The LPS glycolipid is a major component of the outer membrane of gram-negative bac...

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Section: Discussionmentioning

confidence: 96%

Single polysaccharide assembly protein that integrates polymerization, termination, and chain-length quality control

Williams

Ovchinnikova

Koizumi

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…In a number of L-Rha-containing OPSs, synthesis of adaptor is catalysed by rhamnosyltransferase WbbL, which transfers a single L-Rha residue to DGlcNAc-PP-Und (Rubires et al, 1997;Mills et al, 2004). Predicted rhamnosyltransferase Orf5 shares 21 % identity and 38 % similarity to WbbL of Aeromonas hydrophila O1 (Merino et al, 2015) and, thus, may fulfil this function in F. pulveris G3872. However, a role of Orf5 in the formation of a rhamnosidic linkage within the repetitive OPS domain rather than in the adaptor cannot be excluded.…”

Section: Discussionmentioning

confidence: 99%

O antigen of Franconibacter pulverisG3872 (O1) is a 4-deoxy-d-arabino-hexose-containing polysaccharide synthesized by the ABC-transporter-dependent pathway

Wang

Arbatsky

et al. 2016

Microbiology

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Franconibacter (Enterobacter, Cronobacter) pulveris bacteria share several typical characteristics with, and hence pose a challenge for the detection of, Cronobacter sakazakii, an emerging opportunistic pathogen, which can cause severe infections in neonates. A structurally variable O-specific polysaccharide (OPS) called O antigen provides the major basis for the typing of Gram-negative bacteria. We investigated the structure and genetics of the O antigen of F. pulveris G3872 (designated O1). An OPS was isolated by mild alkaline degradation of the LPS, whereas the same polysaccharide and its oligosaccharide fragments were obtained by mild acid degradation. Studies by sugar analysis and NMR spectroscopy showed that the OPS contained D-ribose, L-rhamnose (L-Rha) and a rarely occurring monosaccharide 4-deoxy-Darabino-hexose, and the OPS structure was established. The O-antigen gene cluster of F. pulveris G3872 between JUMPStart and gnd genes includes putative genes for glycosyltransferases, ATP-binding cassette (ABC)-transporter genes wzm and wzt, and genes for the synthesis of L-Rha, but no genes for the synthesis of 4-deoxy-D-arabino-hexose. A mutation test with the wzm gene confirmed that the OPS is synthesized and exported by the ABC-transporter-dependent pathway. A trifunctional transferase was suggested to catalyse formation of two glycosidic linkages and add a methyl group to the non-reducing end of the OPS to terminate the chain elongation. A carbohydrate-binding module that presumably recognizes the terminal methyl-modified monosaccharide was found at the C-terminus of Wzt. Primers specific for F. pulveris G3872 were designed based on the wzm gene, which has potential to be used for identification and detection of the O1 serogroup.

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“…The results reported that FKC vaccine applied to tilapia broodstock gave RPS value 78.26% on day five to the offspring after challenged with A. hydrophila (Sukenda et al, 2017), while lipopolysaccharides were a molecular constituent of Gram-negative bacterial outer membranes that are toxic to fish, making the bacteria become virulent. This molecule is amphiphilic and composed of oligosaccharides, polysaccharides, O-antigen, and lipid-A in the form of acetylglucosamine phosphate fatty acid (Merino et al, 2015). LPS in bacteria are able to provide an immunomodulatory effect on the host (Dehghani et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Efficacy of Aeromonas hydrophila formalin-killed cells and lipopolysaccharides vaccines in maternal immunity of tilapia broodstock and the offspring resistance

Mulyani

Sukenda

Nuryati

2019

Jurnal Akuakultur Indonesia

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The study aimed to analyze the efficacy of Aeromonas hydrophila formalin-killed cells (FKC) and lipopolysaccharides (LPS) vaccines in maternal immunity of tilapia broodstock and the resistance of the offsprings. This study consisted of two consecutive studies. Firstly, the broodstocks were vaccinated with PBS (non-vaccinated), FKC, LPS, and combinations of FKC and LPS. Secondly, the offsprings from both the vaccinated and non‒vaccinated broodstock were vaccinated with the same vaccine for broodstock by immersion method. After vaccination, the offsprings were challenged with pathogenic A. hydrophila. Antibody level and lysozyme were measured in the broodstock, egg, and offsprings. In the first experiment, the result showed that vaccinated broodstock and their offsprings had delivered a significant antibody level and lysozyme activity compared with the control. The highest relative percent survival (RPS) of offspring resulted from the combination of FKC and LPS vaccinated broodstock at 5, 10, and 15 days post-hatching at 85.00%, 75.23%, and 67.56%, respectively. The second experiment showed vaccinated offsprings produced from vaccinated broodstock had a higher RPS than the RPS of vaccinated offsprings which produced from non-vaccinated broodstock. In conclusion, vaccination that using a combination of FKC and LPS vaccine in broodstock potentially improved the maternal immunity and protect their offspring from A. hydrophila infection. Keywords: Aeromonas hydrophila, antibody, maternal immunity, tilapia, vaccination ABSTRAK Penelitian ini bertujuan menganalisis efikasi vaksin formalin-killed cells (FKC) dan lipopolisakarida (LPS) Aeromonas hydrophila pada imunitas maternal induk ikan nila dan ketahanan benih yang dihasilkan. Dalam penelitian ini, terdapat dua penelitian. Pertama, induk ikan nila divaksin dengan PBS (non-vaksinasi), vaksin FKC, LPS, dan kombinasi vaksin FKC dan LPS. Benih selanjutnya diuji ditantang dengan menggunakan patogen A. hydrophila pada 5, 10, dan 15 hari pasca menetas. Kedua, benih yang dihasilkan oleh induk yang telah divaksinasi dan induk yang tidak divaksinasi tersebut divaksin dengan vaksin yang sama seperti pada induk dengan metode imersi. Setelah vaksinasi, keturunan diuji tantang dengan patogen A. hydrophila. Tingkat antibodi dan lisozim diukur pada induk, telur, dan benih. Pada percobaan pertama, hasilnya menunjukkan bahwa induk yang divaksinasi dan benih menunjukkan tingkat antibodi dan aktivitas lisozim yang signifikan dibandingkan dengan kontrol. Persentase kelangsungan hidup relatif (RPS) tertinggi benih yang dihasilkan dari induk yang divaksinasi FKC dan LPS masing-masing pada 5,10, dan 15 hari pascatetas pada 85,00%; 75,23%; dan 67,56%. Percobaan kedua menunjukkan bahwa benih yang divaksinasi dari induk yang divaksin menunjukkan RPS yang lebih baik daripada benih yang divaksinasi dari induk yang tidak divaksin. Vaksinasi menggunakan kombinasi FKC dan LPS pada induk berpotensi meningkatkan kekebalan induk dan melindungi benih dari infeksi A. hydrophila. Kata kunci: Aeromonas hydrophila, antibodi, imunitas induk, nila, vaksinasi

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Molecular and Chemical Analysis of the Lipopolysaccharide from Aeromonas hydrophila Strain AH-1 (Serotype O11)

Cited by 18 publications

References 41 publications

Single polysaccharide assembly protein that integrates polymerization, termination, and chain-length quality control

Single polysaccharide assembly protein that integrates polymerization, termination, and chain-length quality control

O antigen of Franconibacter pulverisG3872 (O1) is a 4-deoxy-d-arabino-hexose-containing polysaccharide synthesized by the ABC-transporter-dependent pathway

Efficacy of Aeromonas hydrophila formalin-killed cells and lipopolysaccharides vaccines in maternal immunity of tilapia broodstock and the offspring resistance

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