title: Cell surface display of Yarrowia lipolytica lipase Lip2p using the cell wall protein YlPir1p, its characterization, and application as a whole-cell biocatalyst.
abstract: The Yarrowia lipolytica lipase Lip2p was displayed on the yeast cell surface via N-terminal fusion variant using cell wall protein YlPir1p. The hydrolytic activity of the lipase displayed on Y. lipolytica cells reached 11,900 U/g of dry weight. However, leakage of enzyme from the cell wall was observed. The calculated number of recombinant enzyme displayed on the cell surface corresponds to approximately 6 × 10(5) molecules per cell, which is close to the theoretical maximum (2 × 10(6) molecules/cell). Furthermore, the leaking enzyme was presented as three N-glycosylated proteins, one of which corresponds to the whole hybrid protein. Thus, we attribute the enzyme leakage to the limited space available on the cell surface. Nevertheless, the surface-displayed lipase exhibited greater stability to short-term and long-term temperature treatment than the native enzyme. Cell-bound lipase retained 74 % of its original activity at 60 °C for 5 min of incubation, and 83 % of original activity after incubation at 50 °C during 5 h. Cell-bound lipase had also higher stability in organic solvents and detergents. The developed whole-cell biocatalyst was used for recycling biodiesel synthesis. Two repeated cycles of methanolysis yielded 84.1 and 71.0 % methyl esters after 33- and 45-h reactions, respectively.
authors.affiliation: Russian National Collection of Industrial Microorganisms (VKPM), State Research Institute of Genetics and Selection of Industrial Microorganisms, 1-st Dorozhniy Pr., 1, Moscow, Russia, 117545, enja_t@mail.ru.
authors.forename: Evgeniya Y
authors.initials: EY
authors.lastname: Yuzbasheva
authors.forename: Tigran V
authors.initials: TV
authors.lastname: Yuzbashev
authors.forename: Natalia I
authors.initials: NI
authors.lastname: Perkovskaya
authors.forename: Elizaveta B
authors.initials: EB
authors.lastname: Mostova
authors.forename: Tatiana V
authors.initials: TV
authors.lastname: Vybornaya
authors.forename: Aleksei V
authors.initials: AV
authors.lastname: Sukhozhenko
authors.forename: Ilya Y
authors.initials: IY
authors.lastname: Toropygin
authors.forename: Sergey P
authors.initials: SP
authors.lastname: Sineoky
mesh_terms: D056804:Biofuels; D002473:Cell Wall / Q000201:enzymology* / Q000235:genetics; D004800:Enzymes, Immobilized / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006868:Hydrolysis; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics
chemical_list: D056804:Biofuels; D004800:Enzymes, Immobilized; D006023:Glycoproteins; D008049:Lipase