Pentose and hexose hydrolyzed from crop straw with acid and enzyme were determined by HPLC.Based on the discussion of corresponding product and enzyme, the metabolic pathway was analyzed.For interdicting lactic acid metabolic pathway and acetic acid metabolic pathway, the parent strain was mutated by soft X-ray of synchronous radiation.In order to improve the balance of H reducing power, the flux ratio of HMP to EMP was modulated by adding citrate.Then the xylulokinase activity was regulated for the co-fermentation of pentose and hexose.The results showed that about 57 g glucose and 11 g xylose were hydrolyzed from 200 g crop straw.Metabolic flux analysis of the parent strain indicated that the flux of succinic acid,lactic acid and acetic acid were 2.547 mmol8226;g-18226;h-1, 0.726 mmol8226;g-18226;h-1 and 0.611 mmol8226;g-18226;h-1, respectively.Lactic acid flux of Ldh mutant strain HF-1 decreased markedly to 0.0296 mmol8226;g-18226;h-1.Acetic acid flux of anti-fluoroacetic mutant strain HF-2 decreased distinctly to 0.100 mmol8226;g-18226;h-1.However, the decrease of acetic acid resulted in not strengthening but weakening the metabolism of succinic acid.By adding citrate the flux ratio of HMP to EMP increased from 1.389∶0.389 to 1.684∶0.330 and thus the H reducing power was better balanced, then the succinic acid flux increased from 3.005 mmol8226;g-18226;h-1 to 3.468 mmol8226;g-18226;h-1 accordingly.Substituting MgCO3 for CaCO3 as neutralizer increased xylulokinase specific activity to 560 U8226;mg-1 from 76 U8226;mg-1, and thus the consumed xylose flux increased from 0.0444 mmol8226;g-18226;h-1 to 0.453 mmol8226;g-18226;h-1 accordingly, then pentose and hexose were co-fermented and the succinic acid yield reached 68.7 g8226;L-1.