Phospholipase D Safe And Efficient Expression In Different Hosts

Phospholipase D (PLD) transphosphatidylation reaction is the most effective way to synthesize a rare natural compounds of high value-added phospholipid.These phospholipids are important raw material for pharmaceuticals, cosmetics and food industry,so large-scale industrial production of phospholipase D has been forced to be resolved. Compared to other PLDs, Streptomyces phospholipase D has a higher reaction activity and a broader substrate selectivity, catalytic structure most closely and higher stability in temperature and the organic medium, which is more suitable for the efficient synthesis of the industry phospholipids and phospholipid derivatives. In this study efficient and safe heterologous and constitutive homologous expression of phospholipase D from soil streptomyces was researched and explored. The E. coli, Pichia pastoris and Yarrowia lipolytica expression system were built to heterologous recombinant expression phospholipase D,which Streptomyces phospholipase D is first expressed in yeast.Then phospholipase D is found that had a toxic effects to E. coli expression host.lt may be an important reason to the low amount of PLD expression in a heterologous host. We constructed two constitutive E. coli-Streptomyces shuttle expression plasmid to effective homologous expression of phospholipase D in Streptomyces lividans TK24.The flask fermentation highest level was to58U/ml with a single expressed strips, Convenient for purification and separation, that provided efficient expression strains for further industrial applications.