Enzymatic properties and expression patterns of five extracellular lipases of Fusarium graminearum in vitro.
Fusarium graminearum is a filamentous fungus that causes devastating diseases on plants of economic importance including maize, wheat, and barley. F. graminearum is able to utilize triglycerides as a carbon source during growth. Extracellular lipases are the preferred enzymes to catalyze the hydrolysis of fats and oils. Lipases are ubiquitous enzymes of considerable physiological significance and industrial potential. Previously, FGL1 was the first described F. graminearum extracellular lipase associated with virulence. We report the biochemical characterization of FGL1 and four new secreted lipases of F. graminearum. The lipase genes of F. graminearum wild-type strain 8/1 were sequenced, cloned and over-expressed in Pichia pastoris. We show that the lipases have their temperature optimum between 30 and 40°C and a pH optimum of ∼7. A broad range of lipase substrates, from C4 to C18 p-nitrophenyl esters, were hydrolyzed efficiently by the lipases, indicating the true lipolytic activity of the enzymes. Expression patterns of these lipases were also analyzed by semiquantitative RT-PCR in F. graminearum cultured in water supplemented with 2% (v/v) wheat germ oil at 28°C. Transcripts of all examined lipases are detectable and the genes are regulated differently under these culture conditions. Our data indicated that F. graminearum possesses a ubiquitous source of secreted lipases, which could be used for industrial intentions. We also provided the foundation of lipase expression in vitro, which is necessary for further characterization.
University of Hamburg, Biocenter Klein Flottbek, Department of Molecular Phytopathology and Genetics, Ohnhorststr. 18, D-22609 Hamburg, Germany; Vietnam Academy of Science and Technology, Institute of Biotechnology, 10600 Hanoi, Viet Nam. Electronic address: namlong@gmail.com.
Long Nam
LN
Nguyen
Vietnam Academy of Science and Technology, Institute of Biotechnology, 10600 Hanoi, Viet Nam.
Tuyet Thi
TT
Dao
University of Hamburg, Biocenter Klein Flottbek, Department of Molecular Phytopathology and Genetics, Ohnhorststr. 18, D-22609 Hamburg, Germany.
Tijana
T
Živković
University of Hamburg, Biocenter Klein Flottbek, Department of Molecular Phytopathology and Genetics, Ohnhorststr. 18, D-22609 Hamburg, Germany.
Markus
M
Fehrholz
University of Hamburg, Biocenter Klein Flottbek, Department of Molecular Phytopathology and Genetics, Ohnhorststr. 18, D-22609 Hamburg, Germany.
Wilhelm
W
Schäfer
University of Hamburg, Biocenter Klein Flottbek, Department of Molecular Phytopathology and Genetics, Ohnhorststr. 18, D-22609 Hamburg, Germany.
Siegfried
S
Salomon
Fusarium graminearum; Gibberella zeae; Hydrolytic enzymes; Lipase