Enzymatic degumming using phospholipase C (PLC) enzymes may be used in environmentally friendly processes with improved oil recovery yields. In this work， phosphatidylinositol-specific phospholipase C (PIPLC) candidates obtained from an in silico analysis were evaluated for oil degumming. A PIPLC from Lysinibacillus sphaericus was shown to efficiently remove phosphatidylinositol from crude oil， and when combined with a second phosphatidylcholine and phosphatidylethanolamine-specific phospholipase C， the three major phospholipids were completely hydrolyzed， providing an extra yield of oil greater than 2.1%， compared to standard methods. A remarkably efficient fed-batch Escherichia coli fermentation process producing ∼14 g/L of the recombinant PIPLC enzyme was developed， which may facilitate the adoption of this cost-effective oil-refining process.