Extracellular cross-linking of xylan and xyloglucan in maize cell-suspension cultures: the role of oxidative phenolic coupling

E M Kerr, S C Fry

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Cell-suspension cultures of maize (Zea mays L.) released soluble extracellular polysaccharides (SEPs) into their medium. Some or all of the SEPs had feruloyl ester groups. Pulse-labelling with [H-3]arabinose was used to monitor changes in the SEPs' M-r (estimated by gel-permeation chromatography) with time after synthesis. Newly released H-3-SEPs were 1.3-1.6 MDa, but between 2 days and 3 days after radiolabelling (in one experiment) or between 5 days and 6 days (in another), the H-3-SEPs abruptly increased to approximate to17 MDa, indicating extensive cross-linking. The cross-linking involved both [H-3]xylan and [H-3]xyloglucan components of the SEPs. The cross-links could be cleaved by alkali, returning the SEPs to their original M-r. In 0.1 M NaOH at 37degreesC, 58% cleavage was effected within 24 h. The requirement for such prolonged alkali treatment indicates that ester-bonded (e.g. diferuloyl) groups were not solely responsible for the cross-linking. Bonds cleaved only by relatively severe alkali could include benzyl ether linkages formed between sugar residues and oxidised phenolics that had quinone methide structures. The ability of alkali to cleave the cross-links was independent of the age of the H-3-SEP molecules. Cross-linking of H-3-SEPs in vivo was delayed (up to approx. 7 days after radiolabelling) by exogenous sinapic acid, chlorogenic acid or rutin-agents predicted to compete with the oxidative coupling of feruloyl-polysaccharides. The cross-linking was promoted by exogenous ferulic acid or L-tyrosine, possibly because these compounds acted as precursors for polysaccharide feruloylation, thus providing additional partner substrates for the oxidative coupling of previously formed H-3-SEPs. The ability of certain phenolics to prevent the cross-linking of H-3-SEPs supports the idea that the cross-linking involved phenolic oxidation.

Original languageEnglish
Pages (from-to)73-83
Number of pages11
Issue number1
Publication statusPublished - May 2004


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