1988 Fiscal Year Final Research Report Summary
Functional Improvement of Food Proteins and Enzymes by Transglutaminase
| Project/Area Number |
62560128
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
製造化学・食品
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| Research Institution | Kyoto University |
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Principal Investigator |
IKURA Koji Kyoto University, Faculty of Agriculture, 農学部, 助手 (00101246)
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| Project Period (FY) |
1987 – 1988
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| Keywords | Use of transglutaminase / Isolation of plasma transglutaminase / Cloning of transglutaminase cDNA / Bacterial production of animal transglutaminase / 組換え型トランスグルタミナーゼ |
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| Research Abstract |
1. Isolation of transglutaminase from bovine plasma: Plasma Factor X222 is a zymogenic transglutaminase comprising two catalytic subunits (a_2) and two noncatalytic subunits (b_2). An immunoadsorbent column containing monoclonal antibody to the noncatalytic subunit provided a one-step purification procedure that isolated the catalytic subunit of bovine plasma Factor XIII from plasma with a high yield.
2. Amino acid sequence of guinea pig liver transglutaminase: The primary structure of guinea pig liver transglutaminase was deduced from its cDNA sequence and compared with that of the catalytic subunit of human plasma Factor XIII. Several regions of strong homology, including the region surrounding the active site cysteine residue, were observed.
3. Amino-terminal processing of guinea pig liver transglutaminase: N- and C-terminal peptides were isolate from protease digests of this enzyme and characterized structurally. These results indicated that this enzyme underwent an N-terminal processing, a removal of the initiator methionine and a subsequent acetylation of newly exposed N-terminal alanine residue.
4. Bacterial production of animal transglutaminase: We constructed an expression plasmid pKTGl containing a cDNA of guinea pig liver transglutaminase between NcoI and PstI sites of an expression vector pKK233-2 and produced the enzyme in E. coli. The catalytic properties were mostly same between recombinant and natural transglutaminases.
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