TU Darmstadt / ULB / TUbiblio

Features of the transglutaminase-activating metalloprotease from Streptomyces mobaraensis DSM 40847 produced in Escherichia coli.

Juettner, Norbert E. ; Classen, Moritz ; Colin, Felix ; Hoffmann, Sascha B. ; Meyners, Christian ; Pfeifer, Felicitas ; Fuchsbauer, Hans-Lothar (2018)
Features of the transglutaminase-activating metalloprotease from Streptomyces mobaraensis DSM 40847 produced in Escherichia coli.
In: Journal of biotechnology, (281)
Article, Bibliographie

Abstract

Transglutaminase from Streptomyces mobaraensis (MTG) is an important enzyme for numerous industrial applications. Recombinant production requires proteolytic activation of the zymogen. The study provides a convenient procedure for the preparation of the transglutaminase-activating metalloprotease (TAMP) in Escherichia coli. In contrast to wtTAMP, rTAMP exhibited the P domain of convertases as molecular mass of 55.7 kDa suggested. Protein integrity was beneficially influenced by 2-5 mM CaCl. Study of pH and temperature optima assigned rTAMP to the neutral metalloproteases, more heat-resistant than Dispase but not thermolysin. Zinc had no inhibiting effect but 3.1 µM EDTA completely reduced activity of 5 nM TAMP. MTG, exceeding concentration of rTAMP by three orders of magnitude, was largely activated within few minutes. The kinetic parameters K (1.31 ± 0.05 mM) and k (135 ± 4.3 s), monitored by isothermal titration calorimetry (ITC), further highlighted catalytic efficiency (103,053 M s) of rTAMP and rapid processing of MTG. ITC even revealed that inhibition of rTAMP by its intrinsic inhibitory protein SSTI was an enthalpy-driven process resulting in K of 199 ± 37.9 nM. The production procedure of rTAMP in E. coli closes the gap between production and application of recombinant MTG and may enhance relevance of MTG-mediated reactions in pharmaceutical processes.

Item Type: Article
Erschienen: 2018
Creators: Juettner, Norbert E. ; Classen, Moritz ; Colin, Felix ; Hoffmann, Sascha B. ; Meyners, Christian ; Pfeifer, Felicitas ; Fuchsbauer, Hans-Lothar
Type of entry: Bibliographie
Title: Features of the transglutaminase-activating metalloprotease from Streptomyces mobaraensis DSM 40847 produced in Escherichia coli.
Language: English
Date: 10 September 2018
Journal or Publication Title: Journal of biotechnology
Issue Number: 281
Abstract:

Transglutaminase from Streptomyces mobaraensis (MTG) is an important enzyme for numerous industrial applications. Recombinant production requires proteolytic activation of the zymogen. The study provides a convenient procedure for the preparation of the transglutaminase-activating metalloprotease (TAMP) in Escherichia coli. In contrast to wtTAMP, rTAMP exhibited the P domain of convertases as molecular mass of 55.7 kDa suggested. Protein integrity was beneficially influenced by 2-5 mM CaCl. Study of pH and temperature optima assigned rTAMP to the neutral metalloproteases, more heat-resistant than Dispase but not thermolysin. Zinc had no inhibiting effect but 3.1 µM EDTA completely reduced activity of 5 nM TAMP. MTG, exceeding concentration of rTAMP by three orders of magnitude, was largely activated within few minutes. The kinetic parameters K (1.31 ± 0.05 mM) and k (135 ± 4.3 s), monitored by isothermal titration calorimetry (ITC), further highlighted catalytic efficiency (103,053 M s) of rTAMP and rapid processing of MTG. ITC even revealed that inhibition of rTAMP by its intrinsic inhibitory protein SSTI was an enthalpy-driven process resulting in K of 199 ± 37.9 nM. The production procedure of rTAMP in E. coli closes the gap between production and application of recombinant MTG and may enhance relevance of MTG-mediated reactions in pharmaceutical processes.

Identification Number: pmid:29981445
Divisions: 10 Department of Biology
10 Department of Biology > Microbiology and Archaea
Date Deposited: 10 Jul 2018 07:40
Last Modified: 28 Aug 2018 06:38
PPN:
Export:
Suche nach Titel in: TUfind oder in Google
Send an inquiry Send an inquiry

Options (only for editors)
Show editorial Details Show editorial Details