Enhanced the Production of Large Heparin Oligosaccharides using Immobilized Microbial Heparinase
Heparin is a naturally occurring anticoagulant and is also used to treat deep vein thrombosis, pulmonary embolism etc. Its major side effects are bleeding and thrombocytopenia. The development of heparin oligosaccharides considered as a better therapeutic agent than unfractionated heparin or disaccharides (dp 2), because of the possibility to reduced non-specific interactions, heterogeneity and have been reported to act as a metastasis in various types of cancers, tumor invasion, inflammation and angiogenesis. Synthesizing longer heparin oligosaccharides takes years of effort and is highly structure-specific. Enzymatic depolymerisation using heparinases is one of the method for production of heparin oligosaccharides. However, uses of heparinases have been hampered due to their undesirable properties such as its efficient catalytic reactions under mild conditions, poor stability and very high cost. Keeping in view the tremendous potential of heparinase for pharmaceutical industries and to make it economical as well as more stable, in this study heparinase was immobilized on CNBr-activated Sepharose, designed immobilized heparinase reactor and and used for production of longer heparin oligosaccharides. Immobilized Heparinase retained high efficiency for depolymerization of heparin at various range of pH (5–8) and temperature (5°–50° C). The optimum parameters i.e., at pH 5.5 and temperature 15°C found better for production of larger proportions of variably sulfated oligosaccharides. Immobilized heparinase had been used10 cycles for degradation of heparin and observed stability in activity on reuse.