Expression, purification, and characterization of histidine-tagged mouse monoglyceride lipase from baculovirus-infected insect cells

Monoglyceride lipase (MGL) has been produced with the baculovirus-insect ce ll system. The mouse MGL cDNA was subcloned into a baculovirus transfer vec tor in frame with a sequence encoding an N-terminal stretch of six histidin e residues. Purification to apparent homogeneity was obtained by nickel-che lating chromatography. The final yield was 3 mg of pure enzymatically activ e MGL per liter of Sf9 cell suspension culture. Analysis by SDS-PAGE and ma ss spectrometry showed that the recombinant histidine-tagged enzyme had the expected molecular mass. With monoolein as substrate, the specific activit y and the apparent K-m were close to those of rat MGL of adipose tissue. (C ) 2000 Academic Press.