Reductive Cleavage Method for Quantitation of Monolignols and Low‐Abundance Monolignol Conjugates

Regner, Matt; Bartuce, Allison; Padmakshan, Dharshana; Ralph, John; Karlen, Steven D.

doi:10.1002/cssc.201800958

Cited by 12 publications

(13 citation statements)

References 33 publications

(67 reference statements)

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“…The derivatization followed by reductive cleavage (DFRC) analysis was performed on maize samples as described for the optimized DFRC protocol (Regner et al, 2018). The maize sample (50 mg) was stirred in a two-dram vial, with PTFE pressure release cap (Chemglass CG-4912-02), in acetyl bromide/acetic acid (1/4 v/v, 3 ml) at 50 • C for 2.5 h. The solvent was removed on a SpeedVac (Thermo Scientific SPD131DDA, 50 • C, 35 min, 1.0 Torr, 35 Torr/min).…”

Section: Derivation Followed By Reductive Cleavage (Dfrc)mentioning

confidence: 99%

“…p-coumarate, from guaiacyl units) (Lu and Ralph, 1999;Regner et al, 2018;Figure 4 and Supplementary Table 7). No significant difference was detected in the means of S-pCA levels of transgenic lines and controls (p = 0.164), showing that the overexpression of ScAT10 had no effect on pCA content in the lignin.…”

Section: Hydroxycinnamate Content Specifically Attached To Gax or Ligninmentioning

confidence: 99%

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Overexpression of a Sugarcane BAHD Acyltransferase Alters Hydroxycinnamate Content in Maize Cell Wall

Fanelli

Rancour²,

Sullivan

et al. 2021

Front. Plant Sci.

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The purification of hydroxycinnamic acids [p-coumaric acid (pCA) and ferulic acid (FA)] from grass cell walls requires high-cost processes. Feedstocks with increased levels of one hydroxycinnamate in preference to the other are therefore highly desirable. We identified and conducted expression analysis for nine BAHD acyltransferase ScAts genes from sugarcane. The high conservation of AT10 proteins, together with their similar gene expression patterns, supported a similar role in distinct grasses. Overexpression of ScAT10 in maize resulted in up to 75% increase in total pCA content. Mild hydrolysis and derivatization followed by reductive cleavage (DFRC) analysis showed that pCA increase was restricted to the hemicellulosic portion of the cell wall. Furthermore, total FA content was reduced up to 88%, resulting in a 10-fold increase in the pCA/FA ratio. Thus, we functionally characterized a sugarcane gene involved in pCA content on hemicelluloses and generated a C4 plant that is promising for valorizing pCA production in biorefineries.

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Section: Derivation Followed By Reductive Cleavage (Dfrc)mentioning

confidence: 99%

Section: Hydroxycinnamate Content Specifically Attached To Gax or Ligninmentioning

confidence: 99%

Overexpression of a Sugarcane BAHD Acyltransferase Alters Hydroxycinnamate Content in Maize Cell Wall

Fanelli

Rancour²,

Sullivan

et al. 2021

Front. Plant Sci.

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“…Extractive-free CWRs for chemical analysis were prepared according to Yamamura et al (2012) and further processed for NMR analysis according to Mansfield et al (2012) and Tobimatsu et al (2013) as described in Materials and Methods S1. Klason lignin assay (Hatfield et al, 1994), thioglycolic acid lignin assay (Suzuki et al, 2009), thioacidolysis (Yamamura et al, 2011;Yue et al, 2012), DFRC Regner et al, 2018), DFRC-Pr (Karlen et al, 2017), determination of cell-wall-bound pCA and FA content (Yamamura et al, 2012), and amorphous and crystalline neutral sugar composition analysis (Lam et al, 2017) were performed largely according to literature methods and as further described in Materials and Methods S1. Analytical parameters and multiple reaction monitoring (MRM) parameters used for DFRC analysis are shown in Tables S6 and S7.…”

Section: Cell Wall Analysismentioning

confidence: 99%

Downregulation of p‐COUMAROYL ESTER 3‐HYDROXYLASE in rice leads to altered cell wall structures and improves biomass saccharification

et al. 2018

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p-Coumaroyl ester 3-hydroxylase (C3'H) is a key enzyme involved in the biosynthesis of lignin, a phenylpropanoid polymer that is the major constituent of secondary cell walls in vascular plants. Although the crucial role of C3'H in lignification and its manipulation to upgrade lignocellulose have been investigated in eudicots, limited information is available in monocotyledonous grass species, despite their potential as biomass feedstocks. Here we address the pronounced impacts of C3'H deficiency on the structure and properties of grass cell walls. C3'H-knockdown lines generated via RNA interference (RNAi)-mediated gene silencing, with about 0.5% of the residual expression levels, reached maturity and set seeds. In contrast, C3'H-knockout rice mutants generated via CRISPR/Cas9-mediated mutagenesis were severely dwarfed and sterile. Cell wall analysis of the mature C3'H-knockdown RNAi lines revealed that their lignins were largely enriched in p-hydroxyphenyl (H) units while being substantially reduced in the normally dominant guaiacyl (G) and syringyl (S) units. Interestingly, however, the enrichment of H units was limited to within the non-acylated lignin units, with grass-specific γ-p-coumaroylated lignin units remaining apparently unchanged. Suppression of C3'H also resulted in relative augmentation in tricin residues in lignin as well as a substantial reduction in wall cross-linking ferulates. Collectively, our data demonstrate that C3'H expression is an important determinant not only of lignin content and composition but also of the degree of cell wall cross-linking. We also demonstrated that C3'H-suppressed rice displays enhanced biomass saccharification.

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“…Besides FA, another esterified HCA is p-coumaric acid (Figure 1), which is mostly reported to be γ-esterified to β-O-4-linkages within the lignin macromolecule, in particular in the PCW of certain Gramineae (Ralph, 2010). Small amounts of FA could also be ester-linked to lignin for which further evidence has yet to be obtained (Regner et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Feruloyl Esterases for Biorefineries: Subfamily Classified Specificity for Natural Substrates

Underlin

Frommhagen

Dilokpimol

et al. 2020

Front. Bioeng. Biotechnol.

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Reductive Cleavage Method for Quantitation of Monolignols and Low‐Abundance Monolignol Conjugates

Abstract: Supporting Information and the ORCID identification number(s) for the author(s) of this article can be found underh ttps://doi.org/10.

Cited by 12 publications

References 33 publications

Overexpression of a Sugarcane BAHD Acyltransferase Alters Hydroxycinnamate Content in Maize Cell Wall

Overexpression of a Sugarcane BAHD Acyltransferase Alters Hydroxycinnamate Content in Maize Cell Wall

Downregulation of p‐COUMAROYL ESTER 3‐HYDROXYLASE in rice leads to altered cell wall structures and improves biomass saccharification

Feruloyl Esterases for Biorefineries: Subfamily Classified Specificity for Natural Substrates

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