We report a novel enzyme-involved approach to improve the extraction of flavonoids from Ginkgo biloba, in which the enzyme is employed not only for cell wall degradation, but also for increasing the solubility of target compounds in the ethanol-water extractant. Penicillium decumbens cellulase, a commercial cell wall-degrading enzyme with high transglycosylation activity, was found to offer far better performance in the extraction than Trichoderma reesei cellulase and Aspergillus niger pectinase under the presence of maltose as the glycosyl donor. TLC, HPLC and MS analysis indicated that P. decumbens cellulase could transglycosylate flavonol aglycones into more polar glucosides, the higher solubility of which led to improved extraction. The influence of glycosyl donor, pH, solvent and temperature on the enzymatic transglycosylation was investigated. For three predominant flavonoids in G. biloba, the transglycosylation showed similar optimal conditions, which were therefore used for the enzyme-assisted extraction. The extraction yield turned to be 28.3 mg/g of dw, 31% higher than that under the pre-optimized conditions, and 102% higher than that under the conditions without enzymes. The utilization of enzymatic bifunctionality described here, naming enzymatic modification of target compounds and facilitation of cell wall degradation, provides a novel approach for the extraction of natural compounds from plants. (C) 2010 Elsevier Inc. All rights reserved.