Characterisation of lignin-carbohydrate complexes (LCCs) of spruce wood (Picea abies L.) isolated with two methods

A method for the quantitative isolation of lignin-carbohydrate complexes (LCCs) in a softwood is presented. The isolation steps involve partial enzymatic hydrolysis of cellulose, subsequent swelling in urea, and quantitative dissolution into four major fractions: (1) a galactoglucomannan LCC containing ∼8% of the wood lignin; (2) a glucane LCC containing ∼4% of the wood lignin; (3) a xylan-lignin-glucomannan network LCC (xylan > glucomannan) containing ∼40% of the wood lignin; and (4) a glucomannan-lignin-xylan network LCC (glucomannan>xylan) containing ∼48% of the wood lignin. Endoglucanase Novozyme 476, with only cellulase activity, and Ecopulp XM, with only xylanase and mannanase activities, were used as an enzymatic tool. From mildly ball-milled wood, all the lignin was isolated as LCCs. As a control, LCC was prepared from partially chlorite-delignified wood meal without ball milling, also in a mild procedure. The results were very similar to those obtained after ball milling. Thus, it can be safely concluded that the formation of new chemical linkages between lignin and carbohydrates during ball milling is improbable. Studies on isolated milled wood lignin (MWL) supported this conclusion and clearly showed that covalent linkages between lignin and carbohydrates are present. The study provide conclusive evidence of covalent linkages between lignin and carbohydrates in the native lignin in wood. It is concluded that carbohydrate-free lignin, i.e., lignin without covalent bonds to carbohydrates, probably cannot be present in spruce wood.