title: NMR-based screening method for transglutaminases: rapid analysis of their substrate specificities and reaction rates.
abstract: Incorporation of inter- or intramolecular covalent cross-links into food proteins with microbial transglutaminase (MTG) improves the physical and textural properties of many food proteins such as tofu, boiled fish paste, and sausage. Other transglutaminases (TGases) are expected to be used in the same way, and also to extend the scope of industrial applications to materials, drugs, and so on. The TGases have great diversity, not only in amino acid sequence and size, but also in their substrate specificities and catalytic activities, and therefore, it is quite difficult to estimate their reactivity. We have developed an NMR-based method using the enzymatic labeling technique (ELT) for simultaneous analysis of the substrate specificities and reaction rates of TGases. It is quite useful for comparing the existing TGases and for screening new TGases or TGases variants. This method has shown that MTG is superior for industrial use because of its lower substrate specificity compared with those of guinea pig liver transglutaminase (GTG) and red sea bream liver transglutaminase (FTG). We have also found that an MTG variant lacking an N-terminal aspartic acid residue has higher activity than that of the native enzyme.
authors.affiliation: Central Research Laboratories, Ajinomoto Company, Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki-shi 210-8681, Japan.
authors.forename: Nobuhisa
authors.initials: N
authors.lastname: Shimba
authors.forename: Kei-ichi
authors.initials: K
authors.lastname: Yokoyama
authors.forename: Ei-ichiro
authors.initials: E
authors.lastname: Suzuki
mesh_terms: D000818:Animals; D002384:Catalysis; D004044:Dietary Proteins / Q000378:metabolism*; D006168:Guinea Pigs; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D021541:Sea Bream; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D004044:Dietary Proteins; D011503:Transglutaminases