Polynucleotide encoding a mutant transglutaminase
First Claim
1. An isolated polynucleotide encoding a mutant MTG having transglutaminase activity wherein the mutant MTG has the amino acid sequence of SEQ ID NO:
- 2 with a mutation, where said mutation is introduced into the sequence by;
a) replacing Ser2 with Arg, Tyr, or Asp;
b) deleting amino acid residue Nos. 1-2 or 1-3 at the N-terminus;
c) replacing amino acid residue Nos. 241-252 with three Gly residues;
ord) replacing amino acid residue Nos. 278-287 with three Gly residues.
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Abstract
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.
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Citations
4 Claims
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1. An isolated polynucleotide encoding a mutant MTG having transglutaminase activity wherein the mutant MTG has the amino acid sequence of SEQ ID NO:
- 2 with a mutation, where said mutation is introduced into the sequence by;
a) replacing Ser2 with Arg, Tyr, or Asp; b) deleting amino acid residue Nos. 1-2 or 1-3 at the N-terminus; c) replacing amino acid residue Nos. 241-252 with three Gly residues;
ord) replacing amino acid residue Nos. 278-287 with three Gly residues. - View Dependent Claims (2, 3, 4)
- 2 with a mutation, where said mutation is introduced into the sequence by;
Specification
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- Resources
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Current AssigneeAjinomoto Co., Inc.
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Original AssigneeAjinomoto Co., Inc.
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InventorsShimba, Nobuhisa, Ishikawa, Kohki, Kashiwagi, Tatsuki, Suzuki, Ei-ichiro, Hirayama, Kazuo, Yokoyama, Keiichi
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Primary Examiner(s)Fronda; Christian L
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Application NumberUS11/505,438Publication NumberTime in Patent Office1,048 DaysField of SearchNoneUS Class Current435/193CPC Class CodesC07K 14/195 from bacteriaC07K 2299/00 Coordinates from 3D structu...C12N 15/102 Mutagenizing nucleic acidsC12N 9/1044 Protein-glutamine gamma-glu...C12Y 203/02013 Protein-glutamine gamma-glu...
