Integrative analysis of the metabolome and transcriptome reveals the molecular regulatory mechanism of isoflavonoid biosynthesis in Ormosia henryi Prain.

Flavonoids are important components of many phytopharmaceuticals, however, most studies on flavonoids and isoflavonoids have been conducted on herbaceous plants of the family Leguminosae, such as soybean, and less attention has been paid to woody plants. To fill this gap, we characterized the metabolome and transcriptome of five plant organs of Ormosia henryi Prain (OHP), a woody Leguminosae plant with great pharmaceutical value. Our results indicate that OHP possesses a relatively high content of isoflavonoids as well as significant diversity, with greater diversity of isoflavonoids in the roots. Combined with transcriptome data, the pattern of isoflavonoid accumulation was found to be highly correlated with differential expression genes. Furthermore, the use of trait-WGCNA network analysis identified OhpCHSs as a probable hub enzyme that directs the downstream isoflavonoid synthesis pathway. Transcription factors, such as MYB26, MYB108, WRKY53, RAV1 and ZFP3, were found to be involved in the regulation of isoflavonoid biosynthesis in OHP. Our findings will be beneficial for the biosynthesis and utilization of woody isoflavonoids.

Real-time imaging reveals that lytic polysaccharide monooxygenase promotes cellulase activity by increasing cellulose accessibility.

BACKGROUND: The high cost of enzymes is one of the key technical barriers that must be overcome to realize the economical production of biofuels and biomaterials from biomass. Supplementation of enzyme cocktails with lytic polysaccharide monooxygenase (LPMO) can increase the efficiency of these cellulase mixtures for biomass conversion. The previous studies have revealed that LPMOs cleave polysaccharide chains by oxidization of the C1 and/or C4 carbons of the monomeric units. However, how LPMOs enhance enzymatic degradation of lignocellulose is still poorly understood. RESULTS: In this study, we combined enzymatic assays and real-time imaging using atomic force microscopy (AFM) to study the molecular interactions of an LPMO [TrAA9A, formerly known as TrCel61A) from Trichoderma reesei] and a cellobiohydrolase I (TlCel7A from T. longibrachiatum) with bacterial microcrystalline cellulose (BMCC) as a substrate. Cellulose conversion by TlCel7A alone was enhanced from 46 to 54% by the addition of TrAA9A. Conversion by a mixture of TlCel7A, endoglucanase, and ß-glucosidase was increased from 79 to 87% using pretreated BMCC with TrAA9A for 72 h. AFM imaging demonstrated that individual TrAA9A molecules exhibited intermittent random movement along, across, and penetrating into the ribbon-like microfibril structure of BMCC, which was concomitant with the release of a small amount of oxidized sugars and the splitting of large cellulose ribbons into fibrils with smaller diameters. The dividing effect of the cellulose microfibril occurred more rapidly when TrAA9A and TlCel7A were added together compared to TrAA9A alone; TlCel7A alone caused no separation. CONCLUSIONS: TrAA9A increases the accessible surface area of BMCC by separating large cellulose ribbons, and thereby enhances cellulose hydrolysis yield. By providing the first direct observation of LPMO action on a cellulosic substrate, this study sheds new light on the mechanisms by which LPMO enhances biomass conversion.