Abstract：Anamperometric glucose dehydrogenase biosensor was reported.It is made up of Toluidine Blue(TB)modified graphite electrode of immerse wax and enzyme film of regenerated silk fibroin w as employed to immobilize glucose dehydrogenase( GDH)、nicotinamide adenine dinucleotide(NAD+). In pH=7. 6 NaOH-KH2PO4 medium, the biosensor has a linear response in the range of 2. 0×10-5～ 5. 4×10-4 mol·L-1 for concentration of glucose.The response time is 20s. The factors affcting the response current of sensor and optimal condition for the determination of glucose have been discussed. The sensor has been used to detect the concentration of glucose in human serum and the result conform to those obtained by enzyme analytical method.