Cloning, expression, crystallization and preliminary X-ray crystallographic analysis of β-ketoacyl-ACP synthase III (FabH) from<i>Xanthomonas oryzae</i>pv.<i>oryzae</i>

Huynh, Kim-Hung; Natarajan, S.; Song, Na; Ngo, Phuong Thuy Ho; Ahn, Yeh Jin; Kim, Jeong Gu; Lee, Byoung Moo; Eo, Yang Dam; Kang, Lin Woo

doi:10.1107/s1744309109009555

Cited by 3 publications

(3 citation statements)

References 18 publications

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“…Little is known of the biosynthetic mechanisms of these fatty acids except that Xanthomonas oryzae pv. oryzae FabD, FabB, FabH and FabV have been expressed and crystallized 26 27 28 29 .…”

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confidence: 99%

The Multiple DSF-family QS Signals are Synthesized from Carbohydrate and Branched-chain Amino Acids via the FAS Elongation Cycle

Zhou

Chen

et al. 2015

Sci Rep

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Members of the diffusible signal factor (DSF) family are a novel class of quorum sensing (QS) signals in diverse Gram-negative bacteria. Although previous studies have identified RpfF as a key enzyme for the biosynthesis of DSF family signals, many questions in their biosynthesis remain to be addressed. In this study with the phytopathogen Xanthomonas campestris pv. campestris (Xcc), we show that Xcc produces four DSF-family signals (DSF, BDSF, CDSF and IDSF) during cell culture, and that IDSF is a new functional signal characterized as cis-10-methyl-2-dodecenoic acid. Using a range of defined media, we further demonstrate that Xcc mainly produces BDSF in the presence of carbohydrates; leucine and valine are the primary precursor for DSF biosynthesis; isoleucine is the primary precursor for IDSF biosynthesis. Furthermore, our biochemical analyses show that the key DSF synthase RpfF has both thioesterase and dehydratase activities, and uses 3-hydroxydedecanoyl-ACP as a substrate to produce BDSF. Finally, our results show that the classic fatty acid synthesis elongation cycle is required for the biosynthesis of DSF-family signals. Taken all together, these findings establish a general biosynthetic pathway for the DSF-family quorum sensing signals.

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“…Little is known of the biosynthetic mechanisms of these fatty acids except that Xanthomonas oryzae pv. oryzae FabD, FabB, FabH and FabV have been expressed and crystallized 26 27 28 29 .…”

mentioning

confidence: 99%

The Multiple DSF-family QS Signals are Synthesized from Carbohydrate and Branched-chain Amino Acids via the FAS Elongation Cycle

Zhou

Chen

et al. 2015

Sci Rep

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“…Although the fatty acid profiles of Xanthomonas have been investigated for taxonomic purposes 22 23 24 , and several fatty acid synthetic enzymes, including FabD, FabB, FabH, and FabV from Xanthomonas oryzae pv. oryzae 25 26 27 28 , have been expressed and crystallized, little is known about the fatty acid biosynthetic pathway in Xanthomonas .…”

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confidence: 99%

Xanthomonas campestris FabH is required for branched-chain fatty acid and DSF-family quorum sensing signal biosynthesis

Dong

et al. 2016

Sci Rep

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Xanthomonas campestris pv. campestris (Xcc), a Gram-negative phytopathogenic bacterium, causes black rot disease of cruciferous vegetables. Although Xcc has a complex fatty acid profile comprised of straight-chain fatty acids and branched-chain fatty acids (BCFAs), and encodes a complete set of genes required for fatty acid synthesis, there is still little known about the mechanism of BCFA synthesis. We reported that expression of Xcc fabH restores the growth of Ralstonia solanacearum fabH mutant, and this allows the R. solanacearum fabH mutant to produce BCFAs. Using in vitro assays, we demonstrated that Xcc FabH is able to condense branched-chain acyl-CoAs with malonyl-ACP to initiate BCFA synthesis. Moreover, although the fabH gene is essential for growth of Xcc, it can be replaced with Escherichia coli fabH, and Xcc mutants failed to produce BCFAs. These results suggest that Xcc does not have an obligatory requirement for BCFAs. Furthermore, Xcc mutants lost the ability to produce cis-11-methyl-2-dodecenoic acid, a diffusible signal factor (DSF) required for quorum sensing of Xcc, which confirms that the fatty acid synthetic pathway supplies the intermediates for DSF signal biosynthesis. Our study also showed that replacing Xcc fabH with E. coli fabH affected Xcc pathogenesis in host plants.

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“…Docking Study. Molecular docking of isolated compounds into the 3D X-ray structure of XooFabH (PDB: 3FK5) 31 was conducted using Discovery Studio 3.5 using a graphical user interface DS-CDocker protocol. The 3D structures of compounds were drawn using ChemBioDraw Ultra 14.0 (Cambridge Soft corporation) with the mol format and were imported into Discovery Studio 3.5.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Discovery of Natural FabH Inhibitors Using an Immobilized Enzyme Column and Their Antibacterial Activity against Xanthomonas oryzae pv. oryzae

Zhao

Xiao

Wang

et al. 2020

J. Agric. Food Chem.

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β-Ketoacyl-acyl carrier protein synthase III (KAS III, FabH) is essential for bacterial fatty acid biosynthesis. Recent studies indicate that FabH can be a potential target for bactericide development. In the present study, an immobilized FabH column was developed and used to screen FabH inhibitors from complex natural product extracts. Combined with HPLC, four secondary metabolites, alternariol (1), altenuisol (2), alterlactone (3), and dehydroaltenusin (4), were site-directed, isolated, and identified from the crude extract of Alternaria alternata ZHJG5. These compounds showed inhibitory activities on FabH of Xanthomonas oryzae pv. oryzae (Xoo) with IC50 values from 29.5 to 74.1 μM and also displayed a varying degree of antibacterial activities against Xoo with minimal inhibitory concentration values from 4 to 64 μg/mL. Molecular modeling was then used to picture how the compounds interact with XooFabH. Two inhibitors, compounds 1 and 3, exhibited significant bactericidal activity against rice bacterial leaf blight with a protective efficiency of 66.2 and 82.5% at the concentration of 200 μg/mL, respectively, suggesting that they could be lead candidates to develop novel bactericides.

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Cloning, expression, crystallization and preliminary X-ray crystallographic analysis of β-ketoacyl-ACP synthase III (FabH) fromXanthomonas oryzaepv.oryzae

Cited by 3 publications

References 18 publications

The Multiple DSF-family QS Signals are Synthesized from Carbohydrate and Branched-chain Amino Acids via the FAS Elongation Cycle

The Multiple DSF-family QS Signals are Synthesized from Carbohydrate and Branched-chain Amino Acids via the FAS Elongation Cycle

Xanthomonas campestris FabH is required for branched-chain fatty acid and DSF-family quorum sensing signal biosynthesis

Discovery of Natural FabH Inhibitors Using an Immobilized Enzyme Column and Their Antibacterial Activity against Xanthomonas oryzae pv. oryzae

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