Thermostable phospholipase C from Thermococcus kodakarensis suitable for oil degumming

MARCHISIO, FIORELA; DI NARDO, LUISINA; VAL, DIEGO SEBASTIÁN; PONCE DE LEON, CAMILA; CERMINATI, SEBASTIÁN; ESPARIZ, MARTÍN; RASIA, RODOLFO MAXIMILIANO; MENZELLA, HUGO GABRIEL; CASTELLI, MARÍA EUGENIA

Congreso; LVIII annual meeting of the Argentine Society for Biochemistry and Molecular Biology Research; 2022

During the last decades, the replacement of chemicals used in industrial processes with enzymaticalternatives has become a general trend. Compared to classical chemical catalysts, enzymes offer highspecificity; work under mild conditions of temperature and pH; avoid the use of toxic and deleterioussubstances and the production of toxic end products providing cleaner technologies. Hence, traditionallyused chemicals have been replaced by novel enzymatic alternatives in a wide variety of industrial-scaleprocesses.Phospholipid removal of crude oils or oil degumming, is the first step of the oil refining process. Enzymaticoil degumming exploits the conversion catalyzed by phospholipases to remove vegetable crude oils’phospholipids. This enzymatic method reduces the gums’ volume and increases the overall oil yield. Athermostable phospholipase would be highly advantageous for industrial oil degumming as oiltreatment at higher temperatures would save energy and increase the recovery of oil by facilitating themixing and gums removal. Hyperthermophiles, predominantly distributed in the domain Archaea, are anunlimited source of thermostable enzymes since they are adapted to high temperatures. In this work, abioinformatic search on genomes of thermophile species was performed in order to identify athermostable phospholipase suitable for the high-temperature oil degumming process. As a result, athermostable phosphatidylcholine (PC) (and phosphatidylethanolamine (PE))- specific phospholipaseC from Thermococcus kodakarensis (TkPLC) was identified. TkPLC was successfully expressed andpurified as a recombinant protein in Pichia pastoris (Komagataella phaffii). The enzyme completelyhydrolyzed PC and PE in crude soybean oil degumming reactions at 80 °C. Thereby, TkPLC is a promisingnew candidate for high-temperature enzymatic degumming, an environmentally improved and moreprofitable industrial scale oil degumming process.