(191v) Cloning and Expression of Heparinase Gene from Raoultella Nx-TZ-3-15

Heparin is a highly sulfated, acidic, linear and complex polysaccharide that belongs to the heparin/heparan sulfate glycosaminoglycans family. Enzymatic depolymerization of heparin by heparinases is promising for production of ultra-low molecular weight heparins (ULMWHs) as anticoagulants, due to its mild reaction conditions and high selectivity. We have screened and isolated a novel heparinase-producing strain from soil samples. The strain was identified as Raoultella sp. by 16S ribosomal DNA identification and stored as Raoultella NX-TZ-3-15. The heparinase from Raoultella NX-TZ-3-15 is an intracellular enzyme. The whole genome of this strain has been sequenced. The heparinase gene was cloned from its genome and expressed in Escherichia coli, using the T7 polymerase and pBENT vector. The results showed that the activity of recombinant heparinase reached 1680 IU/L and the specific activity was 2.47 IU/mg after inducing 8 h at 15^oC.