High-level production of Bacillus cereus phospholipase C in Corynebacterium glutamicum

• A rapid genetic engineering platform was used to evaluate Sec and Tat systems for the secretion of B. Cereus PLC in C. glutamicum to obtain an efficient recombinant strain.
• A fed-batch fermentation process was developed to obtain a final concentration of 5.5 g/L of recombinant PLC.
• The recombinant BC-PLC shows a high efficiency for enzymatic oil degumming.

Enzymatic oil degumming (removal of phospholipids) using phospholipase C (PLC) is a well-established and environmentally friendly process for vegetable oil refining. In this work, we report the production of recombinant Bacillus cereus PLC in Corynebacterium glutamicum ATCC 13869 in a high cell density fermentation process and its performance in soybean oil degumming. A final concentration of 5.5 g/L of the recombinant enzyme was achieved when the respective gene was expressed from the tac promoter in a semi-defined medium. After treatment with trypsin to cleave the propeptide, the mature enzyme completely hydrolyzed phosphatidylcholine and phosphatidylethanolamine, which represent 70% of the phospholipids present in soybean oil. The results presented here show the feasibility of using B. cereus PLC for oil degumming and provide a manufacturing process for the cost effective production of this enzyme.