Entacapone improves saccharification without affecting lignin and maize growth: An in silico, in vitro, and in vivo approach.

Caffeate 3-O-methyltransferase (COMT) catalyzes the methylation of the 3-hydroxyl group of caffeate to produce ferulate, an important precursor of the lignin biosynthesis. As a crucial drawback for biofuel production, lignin limits the enzymatic hydrolysis of polysaccharides to result in fermentable sugars. We hypothesized that a controlled inhibition of maize COMT can be an efficient approach to reduce ferulate and lignin, thus improving the saccharification process. First, we applied in silico techniques to prospect potential inhibitors of ZmaysCOMT, and the nitrocatechol entacapone was selected. Second, in vitro assays confirmed the inhibitory effect of entacapone on maize COMT. Finally, in vivo experiments revealed that entacapone reduced the contents of cell-wall-esterified hydroxycinnamates and increased saccharification of stems (18%) and leaves (70%), without negatively affecting maize growth and lignin biosynthesis. This non-genetically modified approach can be an alternative strategy to facilitate the enzymatic hydrolysis of biomass polysaccharides and increase saccharification for bioethanol production.

Exogenous application of rosmarinic acid improves saccharification without affecting growth and lignification of maize.

Biomimetically incorporated into the lignin structure, rosmarinic acid improves in vitro maize cell wall saccharification; however, no in planta studies have been performed. We hypothesized that rosmarinic acid, itself, could inducer saccharification without disturbing plant growth. Its effects on growth, enzymes of the phenylpropanoid pathway, lignin, monomeric composition, and saccharification of maize were evaluated. In a short-term (24 h) exposure, rosmarinic acid caused deleterious effects on maize roots, inhibiting the first enzymes of the phenylpropanoid pathway, phenylalanine ammonia-lyase and tyrosine ammonia-lyase, altering lignin composition and slightly increasing saccharification. In a long-term (14 d) exposure, rosmarinic acid increased saccharification of maize stems by about 50% without any deleterious effects on plant growth, the phenylpropanoid pathway and lignin formation. This demonstrated that exogenous application of rosmarinic acid on maize plants improved saccharification, and represented an interesting approach in facilitating enzymatic hydrolysis of biomass polysaccharides and increasing bioethanol production.