title: High-level production of Bacillus cereus phospholipase C in Corynebacterium glutamicum.
abstract: 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 5.5g/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.
authors.affiliation: Genetic Engineering & Fermentation Technology, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario-IPROBYQ-Conicet, Suipacha 531, Rosario 2000, Argentina.
authors.forename: Pablo
authors.initials: P
authors.lastname: Ravasi
authors.affiliation: Genetic Engineering & Fermentation Technology, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario-IPROBYQ-Conicet, Suipacha 531, Rosario 2000, Argentina.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Braia
authors.affiliation: Genetic Engineering & Fermentation Technology, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario-IPROBYQ-Conicet, Suipacha 531, Rosario 2000, Argentina.
authors.forename: Florencia
authors.initials: F
authors.lastname: Eberhardt
authors.affiliation: Keclon, S.A. Tucuman 7180, Rosario 2000, Argentina.
authors.forename: Claudia
authors.initials: C
authors.lastname: Elena
authors.affiliation: Genetic Engineering & Fermentation Technology, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario-IPROBYQ-Conicet, Suipacha 531, Rosario 2000, Argentina.
authors.forename: Sebastián
authors.initials: S
authors.lastname: Cerminati
authors.affiliation: Genetic Engineering & Fermentation Technology, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario-IPROBYQ-Conicet, Suipacha 531, Rosario 2000, Argentina; Keclon, S.A. Tucuman 7180, Rosario 2000, Argentina.
authors.forename: Salvador
authors.initials: S
authors.lastname: Peirú
authors.affiliation: Genetic Engineering & Fermentation Technology, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario-IPROBYQ-Conicet, Suipacha 531, Rosario 2000, Argentina; Keclon, S.A. Tucuman 7180, Rosario 2000, Argentina.
authors.forename: Maria Eugenia
authors.initials: ME
authors.lastname: Castelli
authors.affiliation: Genetic Engineering & Fermentation Technology, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario-IPROBYQ-Conicet, Suipacha 531, Rosario 2000, Argentina; Keclon, S.A. Tucuman 7180, Rosario 2000, Argentina. Electronic address: hmenzella@fbioyf.unr.edu.ar.
authors.forename: Hugo G
authors.initials: HG
authors.lastname: Menzella
mesh_terms: D001409:Bacillus cereus / Q000201:enzymology*; D061249:Batch Cell Culture Techniques; D002452:Cell Count; D002851:Chromatography, High Pressure Liquid; D048230:Corynebacterium glutamicum / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D005285:Fermentation; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000378:metabolism; D010738:Type C Phospholipases / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D004247:DNA; D010738:Type C Phospholipases
keywords: Corynebacterium glutamicum; Enzymatic degumming; Green chemistry; Phospholipase C; Synthetic biology