To reveal the structural characteristics and physicochemical properties of perennial ryegrass lignin, sequential alkali extractions or double ball-milling and enzymatic hydrolysis on the basis of ultrasonic and hydrothermal pretreatments were proposed in this study. Results revealed that sequential alkali extractions released 89.4% of original lignin from the ryegrass cell walls and 0.75-4.16% of associated carbohydrates as compared to the double ball-milling and enzymatic hydrolysis (96.0% and 18.39%). It was observed that the two types of lignin prepared were SGH-type and had different amounts of p-coumarates and ferulates, and primarily consisted of β-O-4' linkages combined with minor amounts of β-β' and β-5' linkages. Besides, alkali-soluble lignins exhibited relatively fewer β-O-4' linkages, higher S/G ratios and H-type units, and abundant phenolic OH groups as compared to the double enzymatic lignin. Overall, the deeper investigation of the lignin structure of ryegrass will provide useful information for the efficient utilization of lignin macromolecules in biorefineries.
Keywords: Alkali-soluble lignins; Double enzymatic lignin; Perennial ryegrass.
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