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AssigneeAjinomoto Co., Inc.
InventorsTatsuki Kashiwagi, Nobuhisa Shimba, Kohki Ishikawa, Ei-Ichiro Suzuki, Keiichi Yokoyama, Kazuo Hirayama
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The present invention relates to a method for designing and preparing mutant transglutaminases on the basis of the three-dimensional structure of MTG derived from Streptoverticillium mobaraense (MTG), and the mutant MTG thus prepared. The present invention provides a method for modifying MTG on the basis of the three-dimensional structure, and transglutaminase having reactivity on the substrate improved by the method. In the present invention, the binding site of MTG for the substrate is extrapolated based on the three-dimensional structure obtained by X-ray crystal structure analysis of MTG crystals, and the mutant transglutaminases are designed and produced by replacing, inserting or deleting amino acid residues positioned at the substrate-binding site of the transglutaminase.
A method for designing and preparing a mutant transglutaminase by extrapolating the binding site of a transglutaminase derived from Streptoverticillium mobaraense (MTG) for the substrate on the basis of the three-dimensional structure obtained by X-ray crystal structure analysis and replacing, inserting or deleting amino acid residues positioned at the extrapolated substrate-binding site of the transglutaminase.
A mutant MTG (excluding a mutant MTG wherein Asp1is deleted) having modified substrate specificity, in which at least one of the following positions (residues positioned within 20 Å from active residue Cys64 and exposed on the surface of the extrapolated binding site for acyl acceptor) in the MTG amino acid sequence according to SEQ ID No:
A mutant transglutaminase having transglutaminase activity and modified substrate specificity, wherein when the alignment of the sequence of said mutant transglutaminase with the MTG amino acid sequence according to SEQ ID NO:
A mutant transglutaminase having transglutaminase activity and modified substrate specificity, wherein when the alignment of the three-dimensional structure with the MTG amino acid sequence according to SEQ ID NO:
A monoclinic MTG crystals belonging to P21 space group.
Dependencies are inferred from the claim text.