Ajinomoto Co., Ltd.

Nobuhisa Shimba

Kohki Ishikawa

Eiichiro Suzuki

Keiko Gondoh

Keiichi Yokoyama

Naoyuki Fukuchi

Morikazu Kito

Naoyuki Yamada

Takashi Kayahara

Ei-ichiro Suzuki

Fumie Futaki

Osamu Kurahashi

Yasuhisa Asano

Yasuhiro Mihara

Hisashi Kawasaki

Tatsuki Kashiwagi

Megumi Shimaoka

Kazuo Hirayama

Tohru Kouda

Rie Kozutsumi

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(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.

Polynucleotide encoding a mutant transglutaminase

A method for producing a protein having an antithrombotic activity, which comprises replacing, in a protein that has an amino acid sequence having a homology of not less than 30% to the amino acid sequence of SEQ ID NO: 1 and forms a higher order structure composed of a first &#x3b2; strand (&#x3b2;1), a first a helix (&#x3b1;1), a second &#x3b1; helix (&#x3b1;2), a second &#x3b2; strand (&#x3b2;2), a loop, a third &#x3b2; strand (&#x3b2;3), a fourth &#x3b2; strand (&#x3b2;4) and a fifth &#x3b2; strand (&#x3b2;5) in this order from the amino terminus, at least one amino acid residue in a region from &#x3b1;2 to &#x3b2;2 and/or a region from &#x3b2;3 to &#x3b2;4 so that electric charge of the amino acid residue is changed towards positive direction.

Protein having antithrombotic activity and method for producing the same

There is provided a mutant nucleoside-5'-phosphate producing enzyme with improved nucleoside-5&#x2032;-phosphate producing ability of a nucleoside-5&#x2032;-phosphate producing enzyme that has transphosphorylation activity and/or phosphatase activity and has one Lys residue, two Arg residues and two His residues with distances between their C&#x3b1;'s within a particular ranges and a space around them allowing a binding of a nucleoside, of which mutation is designed by using the crystal structure of EB-AP.

Variant nucleoside-5'-phosphate producing enzyme

Method for isotope labeling of protein with enzyme

A method for producing a protein having an antithrombotic activity, which comprises replacing, in a protein that has an amino acid sequence having a homology of not less than 30% to the amino acid sequence of SEQ ID NO: 1 and forms a higher order structure composed of a first &bgr; strand (&bgr;1), a first &agr; helix (&agr;1), a second &agr; helix (&agr;2), a second &bgr; strand (&bgr;2), a loop, a third &bgr; strand (&bgr;3), a fourth &bgr; strand (&bgr;4) and a fifth &bgr; strand (&bgr;5) in this order from the amino terminus, at least one amino acid residue in a region from &agr;2 to &bgr;2 and/or a region from &bgr;3 to &bgr;4 so that electric charge of the amino acid residue is changed towards positive direction.

Growing community of inventors

Kawasaki, Japan

Nobuhisa Shimba