The impact of xylan on the biosynthesis and structure of extracellular lignin produced by a Norway spruce tissue culture

Abstract

In order to develop more economic uses of lignin, greater knowledge regarding its native structure is required. This can inform the development of optimized extraction methods that preserve desired structural properties. Current extraction methods alter the polymeric structure of lignin, leading to a loss of valuable structural groups or the formation of new non‐native ones. In this study, Norway spruce (Picea abies) tissue‐cultured cells that produce lignin extracellularly in a suspension medium were employed. This system enables the investigation of unaltered native lignin, as no physicochemical extraction steps are required. For the first time, this culture was used to investigate the interactions between lignin and xylan, a secondary cell wall hemicellulose, and to study the importance of lignin–carbohydrate complexes (LCCs) on the polymerization and final structure of extracellular lignin (ECL). This has enabled us to study the impact of xylan on monolignol composition and structure of the final lignin polymer. We find that the addition of xylan to the solid culture medium accelerates cell growth and impacts the ratio of monolignols in the lignin. However, the presence of xylan in the lignin polymerization environment does not significantly alter the structural properties of lignin as analyzed by two‐dimensional nuclear magnetic resonance (NMR) spectroscopy and size exclusion chromatography (SEC). Nevertheless, our data indicate that xylan can act as a nucleation point, leading to more rapid lignin polymerization, an important insight into biopolymer interactions during cell wall synthesis in wood. Lignin structure and interactions with a secondary cell wall hemicellulose were investigated in a model cell culture: we found that the polymerization and final structure of lignin are altered when the hemicellulose is present during cell growth and monolignol production. The physicochemical interactions between lignin and xylan partly define the extractability and utility of native lignin in high value applications, so this work has implications for lignin extraction as well as fundamental plant biology.

Modification of xylan in secondary walls alters cell wall biosynthesis and wood formation programs

“…Whereas one mechanism of this interdependence has been proposed via xylan acting as a nucleation point for the polymerization of monolignols (Sapouna et al, 2023) our data point to a different possibility. First, we observed a severe decrease in mono- and oligolignols as well as phenolic glycosides suggesting severely decreased lignin monomer biosynthesis in xylanase-expressing lines.…”

“…In poplar, Ruel et al (2006) proposed that hemicellulose-lignin covalent linkage serves as an anchor for lignin polymerization. The extracellular lignin analysis in spruce cell cultures suggests that exogenously supplied xylan can act as a nucleation center for lignin polymerization, depending on the amount and solubility of the xylan provided (Sapouna et al, 2023).…”