Purification and Characterization of UDP-Arabinopyranose Mutase from<i>Chlamydomonas reinhardtii</i>

Kotani, Ayana; Tsuji, Maki; Azama, Yasushi; Ishii, Tadashi; Takeda, Takumi; Yamashita, Tetsuro; Shimojima, Mie; Konishi, Teruko

doi:10.1271/bbb.130302

Cited by 11 publications

(12 citation statements)

References 25 publications

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“…UDP- D -xylose is either converted to 1,4-β- D -xylan by a xylan synthase (XS) or into the arabinose precursor UDP- L -Ara p (Rennie and Scheller, 2014). UDP- L -Ara p is converted to UDP- L -Ara f by UAM and then recruited into arabinose or arabinoxylan (Konishi et al, 2011; Rautengarten et al, 2011; Kotani et al, 2013; Rennie and Scheller, 2014; Rancour et al, 2015). We propose that the reduction in available UDP- L -Ara f caused by PvUAM -RNAi results in an increase of UDP- D -xylose with a corresponding reduction in arabinoxylan branching.…”

Section: Discussionmentioning

confidence: 99%

“…UDP-arabinopyranose mutase (UAM) converts UDP-arabinose (pyranose-form, Ara p ) to UDP-Ara f (Konishi et al, 2007, 2010). UAM orthologs are found in some microalgae and land plants in which they comprise a small gene family (Kotani et al, 2013). Interestingly, UAM can also reversibly glycosylate itself in the presence of UDP-sugars, such as UDP-glucose, UDP-galactose, and UDP-xylose [hence the name RGP (reversible glycosylated polypeptide); Dhugga et al, 1991; Konishi et al, 2007, 2010; Rautengarten et al, 2011].…”

Section: Introductionmentioning

confidence: 99%

“…The role of UAM as an RGP is not well understood in the context of cell wall and glycan formation. It was hypothesized that the RGP function of UAM may regulate the internal balance of UDP-sugars in the cell or compete for the formation of UDP-Ara f , and this hypothesis has been explored in various taxa including Arabidopsis , algae, Brachypodium , and rice (Konishi et al, 2011; Rautengarten et al, 2011; Kotani et al, 2013; Rancour et al, 2015). However, UAM’s potential role in recalcitrance has never been examined nor manipulated in any bioenergy feedstock.…”

Section: Introductionmentioning

confidence: 99%

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Downregulation of a UDP-Arabinomutase Gene in Switchgrass (Panicum virgatum L.) Results in Increased Cell Wall Lignin While Reducing Arabinose-Glycans

et al. 2016

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Background: Switchgrass (Panicum virgatum L.) is a C4 perennial prairie grass and a dedicated feedstock for lignocellulosic biofuels. Saccharification and biofuel yields are inhibited by the plant cell wall’s natural recalcitrance against enzymatic degradation. Plant hemicellulose polysaccharides such as arabinoxylans structurally support and cross-link other cell wall polymers. Grasses predominately have Type II cell walls that are abundant in arabinoxylan, which comprise nearly 25% of aboveground biomass. A primary component of arabinoxylan synthesis is uridine diphosphate (UDP) linked to arabinofuranose (Araf). A family of UDP-arabinopyranose mutase (UAM)/reversible glycosylated polypeptides catalyze the interconversion between UDP-arabinopyranose (UDP-Arap) and UDP-Araf.Results: The expression of a switchgrass arabinoxylan biosynthesis pathway gene, PvUAM1, was decreased via RNAi to investigate its role in cell wall recalcitrance in the feedstock. PvUAM1 encodes a switchgrass homolog of UDP-arabinose mutase, which converts UDP-Arap to UDP-Araf. Southern blot analysis revealed each transgenic line contained between one to at least seven T-DNA insertions, resulting in some cases, a 95% reduction of native PvUAM1 transcript in stem internodes. Transgenic plants had increased pigmentation in vascular tissues at nodes, but were otherwise similar in morphology to the non-transgenic control. Cell wall-associated arabinose was decreased in leaves and stems by over 50%, but there was an increase in cellulose. In addition, there was a commensurate change in arabinose side chain extension. Cell wall lignin composition was altered with a concurrent increase in lignin content and transcript abundance of lignin biosynthetic genes in mature tillers. Enzymatic saccharification efficiency was unchanged in the transgenic plants relative to the control.Conclusion: Plants with attenuated PvUAM1 transcript had increased cellulose and lignin in cell walls. A decrease in cell wall-associated arabinose was expected, which was likely caused by fewer Araf residues in the arabinoxylan. The decrease in arabinoxylan may cause a compensation response to maintain cell wall integrity by increasing cellulose and lignin biosynthesis. In cases in which increased lignin is desired, e.g., feedstocks for carbon fiber production, downregulated UAM1 coupled with altered expression of other arabinoxylan biosynthesis genes might result in even higher production of lignin in biomass.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Downregulation of a UDP-Arabinomutase Gene in Switchgrass (Panicum virgatum L.) Results in Increased Cell Wall Lignin While Reducing Arabinose-Glycans

et al. 2016

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“…In sharp contrast with the plant UGE I family, close homologues to Arabidopsis MUR4 can be found not only in gymnosperms, ferns, and mosses, but also in green algae. Although the biochemical properties of MUR4 homologues in green algae have not been determined, the existence of l -Ara-containing glycoprotein and UAM catalyzing subsequent conversion of UDP- l -Ara p to UDP- l -Ara f strongly suggests their role as UXE (Bollig et al 2007 ; Kotani et al 2013 ). Together with algal homologues, MUR4 and MUR4 homologues in land plants form a subclade apart from plant the UGE I and II families, suggesting that MUR4 is a highly conserved old enzyme in plants.…”

Section: Origin Of Udp-xyl 4-epimerasementioning

confidence: 99%

Metabolism of l-arabinose in plants

et al. 2016

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l-Arabinose (l-Ara) is a plant-specific sugar accounting for 5–10 % of cell wall saccharides in Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa). l-Ara occurs in pectic arabinan, rhamnogalacturonan II, arabinoxylan, arabinogalactan-protein (AGP), and extensin in the cell walls, as well as in glycosylated signaling peptides like CLAVATA3 and small glycoconjugates such as quercetin 3-O-arabinoside. This review focuses on recent advances towards understanding the generation of l-Ara and the metabolism of l-Ara-containing molecules in plants.Electronic supplementary materialThe online version of this article (doi:10.1007/s10265-016-0834-z) contains supplementary material, which is available to authorized users.

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“…This activity was initially identified through the fractionation of rice-seedling extracts and monitoring of UDP-arabinopyranose mutase activity (Konishi et al, 2007). Enzymes with UAM activity have been identified in rice, mung bean, Arabidopsis thaliana, green algae and wheat (Hsieh et al, 2016;Konishi et al, 2007;Kotani et al, 2013;Rautengarten et al, 2011), indicating that this activity is evolutionarily conserved within the plant kingdom. While the biological function of UAMs remained elusive until recently, studies now indicate a role in cell-wall biosynthesis; UAM knockdown transformants with decreased cell-wall arabinose content were generated in A. thaliana (Rautengarten et al, 2011) and UAM gene downregulation results in decreased cell-wall arabinose in switchgrass (Willis et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

X-ray diffraction analysis andin vitrocharacterization of the UAM2 protein fromOryza sativa

et al. 2017

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The role of seemingly non-enzymatic proteins in complexes interconverting UDP-arabinopyranose and UDP-arabinofuranose (UDP-arabinosemutases; UAMs) in the plant cytosol remains unknown. To shed light on their function, crystallographic and functional studies of the seemingly non-enzymatic UAM2 protein fromOryza sativa(OsUAM2) were undertaken. Here, X-ray diffraction data are reported, as well as analysis of the oligomeric state in the crystal and in solution. OsUAM2 crystallizes readily but forms highly radiation-sensitive crystals with limited diffraction power, requiring careful low-dose vector data acquisition. Using size-exclusion chromatography, it is shown that the protein is monomeric in solution. Finally, limited proteolysis was employed to demonstrate DTT-enhanced proteolytic digestion, indicating the existence of at least one intramolecular disulfide bridge or, alternatively, a requirement for a structural metal ion.

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Purification and Characterization of UDP-Arabinopyranose Mutase fromChlamydomonas reinhardtii

Cited by 11 publications

References 25 publications

Downregulation of a UDP-Arabinomutase Gene in Switchgrass (Panicum virgatum L.) Results in Increased Cell Wall Lignin While Reducing Arabinose-Glycans

Downregulation of a UDP-Arabinomutase Gene in Switchgrass (Panicum virgatum L.) Results in Increased Cell Wall Lignin While Reducing Arabinose-Glycans

Metabolism of l-arabinose in plants

X-ray diffraction analysis andin vitrocharacterization of the UAM2 protein fromOryza sativa

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