tailieunhanh - Báo cáo khoa học: Transglutaminase from Streptomyces mobaraensis is activated by an endogenous metalloprotease

Streptomyces mobaraensis secretes a Ca 2+ -independent transglutaminase (TGase) that is activated by removing an N-terminal peptide from a precursor protein during sub-merged culture in a complex medium [Pasternack, R., Dorsch, S., Otterbach, J. T., Robenek, I. R., Wolf, S. & Fuchsbauer, . (1998)Eur. J. Biochem. 257, 570–576]. However, an activating protease could not be identified, probably because of the presence of a 14-kDa protein (P14 ) belonging to theStreptomycessubtilisin inhibitor family. . | Eur. J. Biochem. 270 3214-3222 2003 FEBS 2003 doi Transglutaminase from Streptomyces mobaraensis is activated by an endogenous metalloprotease J. Zotzel P. Keller and . Fuchsbauer Fachbereich Chemie- und Biotechnologie Fachhochschule Darmstadt Germany Streptomyces mobaraensis secretes a Ca2 -independent transglutaminase TGase that is activated by removing an N-terminal peptide from a precursor protein during submerged culture in a complex medium Pasternack R. Dorsch S. Otterbach J. T. Robenek I. R. Wolf S. Fuchsbauer . 1998 Eur. J. Biochem. 257 570-576 . However an activating protease could not be identified probably because of the presence of a 14-kDa protein P14 belonging to the Streptomyces subtilisin inhibitor family. In contrast if the microorganism was allowed to grow on a minimal medium several soluble proteases were extracted among them the TGase-activating protease TAMEP . TAMEP was purified by seuuential chromalograpliy on DEAE- and Arg-Sepharose and used to determine the cleavage site of TGase. It was clearly shown that the peptide bond between Phe -4 and Ser -5 was hydrolyzed indicating that at least one additional peptidase is necessary to complete TGase processing even if TAMEPcleavage was sufficient to obtain total activity. Sequence analysis from the N-terminus of TAMEPrevealed the close relationship to a zinc endo-protease from S. griseus. The S. griseus protease differs from other members of the M4 protease family such as thehmolysin in that it may be inhibited by the Strepto-myces subtilisin inhibitor. P14 likewise inhibits TAMEP in approximately equimolar concentrations suggesting its important role in regulating TGase activity. Keywords Streptomyces mobaraensis transglutaminase transglutaminase activation transglutaminase activating metalloprotease Streptomyces subtilisin inhibitor. Streptomycetes are Gram-positive filamentous soil bacteria that exhibit a complex life-cycle comprising a number of .