Abstract: E.coli engineering-bacterium that expressed 1,3-propanediol dehydrogenase(PDH) gene from Klebsiella.p was cultured in 5 L fermentor at 30℃，pH7.0，200 r/min，microaerobic condition for 20h，after the fermented cells were crushed and separated，the acquired sample was then one-step purified to homogeneity by affinity chromatography with HisTrapHP column，this purification process resulted in a 2.94 fold purification of PDH with 50% final yield. One clarity strip was acquired and the relative molecular weight of expression PDH subunits was determined to be approximately 41 kDa using SDS-PAGE. The purified PDH was immobilized by reformative sol-gel (composite gelation) and the immobilized effect was affected by the concentration of tetraethyl orthosilicate (TEOS)，alginate (ALG)，CaCl2 and volume ratio of ALG and TEOS. The result showed that optimized immobilized condition was ρ(TEOS)=20 g/L，ρ(ALG)=30 g/L，ρ(CaCl2)=40 g/L，V(ALG)∶V(TEOS)=3∶1. The ALG-SiO2 composite gelation of PDH was stored for 60 days，and immobilized enzyme activity was preserved 80%. When it reacted for batchs，the immobilized enzyme activity was preserved 70.3% after it reacted for 3 batchs，but was preserved 29.8% after it reacted for 6 batchs.