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In this study, birch wood chips were treated in one-step ball milling (BM) hydrolysis with dilute citric acid and fungus Penicillium simplicissimum at room temperature and atmospheric pressure. An efficient conversion process for the production of fermentable sugars from woody biomasses using wet BM system was developed, in which wood lignocellulose was hydrolyzed into reducing sugars with the total yield of 245.3 mg/g wood. The concentrations of several major substances in the hydrolyzate were discussed in detail. The yields of the monomeric sugars were notably increased in the presence of fungus P. simplicissimum. Corresponding structure transformations before and after milling were analyzed by X-ray diffraction, UV spectroscopy, transmission Fourier transform infrared spectroscopy, and environmental scanning electron microscopy clearly indicated that this combined treatment could be attributed to the crystalline and chemical structure changes of wood lignocellulose during BM. When compared with traditional method of BM, this work showed a more simple, novel, and environmental friendly way in mechanochemical treatment of lignocellulosic biomass, especially woody biomass. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013 [ABSTRACT FROM AUTHOR]