{"id":3131,"date":"2012-01-23T13:49:55","date_gmt":"2012-01-23T04:49:55","guid":{"rendered":"http:\/\/www.ag.kagawa-u.ac.jp?post_type=research&p=3131"},"modified":"2024-05-09T11:40:47","modified_gmt":"2024-05-09T02:40:47","slug":"%e6%ab%bb%e5%ba%ad%e3%80%80%e6%98%a5%e5%bd%a6%ef%bc%88%e9%85%b5%e7%b4%a0%e5%b7%a5%e5%ad%a6%ef%bc%89","status":"publish","type":"research","link":"https:\/\/www.ag.kagawa-u.ac.jp\/?research=%e6%ab%bb%e5%ba%ad%e3%80%80%e6%98%a5%e5%bd%a6%ef%bc%88%e9%85%b5%e7%b4%a0%e5%b7%a5%e5%ad%a6%ef%bc%89","title":{"rendered":"\u6afb\u5ead\u3000\u6625\u5f66"},"content":{"rendered":"
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\u9818\u3000\u57df\u3000\u540d\u3000\u3000\u751f\u547d\u6a5f\u80fd\u79d1\u5b66<\/mark><\/strong><\/h2>\n\n\n\n

\u6559\u54e1\u6c0f\u540d\u3000\u3000 \u6afb\u3000\u5ead\u3000\u6625\u3000\u5f66<\/mark><\/strong><\/h2>\n<\/div>\n\n\n\n
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\u7814\u7a76\u5206\u91ce\u3000\u9175\u7d20\u5de5\u5b66<\/mark><\/strong><\/h2>\n<\/div>\n<\/div>\n\n\n\n
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\u7814\u7a76\u30ad\u30fc\u30ef\u30fc\u30c9\uff1a\u8d85\u597d\u71b1\u83cc\uff0c\u30a2\u30fc\u30ad\u30a2\uff0c\u8010\u71b1\u6027\u9175\u7d20\uff0cX\u7dda\u7d50\u6676\u69cb\u9020\u89e3\u6790<\/strong><\/p>\n<\/div>\n\n\n\n

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\u6700\u8fd1\u306e\u7814\u7a76\u8ab2\u984c<\/strong><\/h2>\n\n\n\n

\uff11\uff0e\u69cb\u9020\u60c5\u5831\u306b\u57fa\u3065\u304f\u65b0\u898fD-\u30a2\u30df\u30ce\u9178\u8131\u6c34\u7d20\u9175\u7d20\u306e\u5275\u88fd<\/strong><\/h2>\n\n\n\n

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\uff12\uff0e\u8d85\u597d\u71b1\u30a2\u30fc\u30ad\u30a2\u7531\u6765\u8131\u6c34\u7d20\u9175\u7d20\u304c\u793a\u3059\u7279\u7570\u7684\u306a\u88dc\u9175\u7d20\u7d50\u5408\u69d8\u5f0f<\/strong><\/h2>\n\n\n\n
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\u3000\u30e1\u30c1\u30aa\u30cb\u30f3\u3001\u30b9\u30ec\u30aa\u30cb\u30f3\u3001\u30a4\u30bd\u30ed\u30a4\u30b7\u30f3\u306f\u5fae\u751f\u7269\u3084\u690d\u7269\u306b\u306e\u307f\u5b58\u5728\u3059\u308b\u4ee3\u8b1d\u7d4c\u8def\u3088\u3063\u3066\u3001\u30a2\u30b9\u30d1\u30e9\u30ae\u30f3\u9178\u304b\u3089\u751f\u5408\u6210\u3055\u308c\u308b\u3002\u3053\u306e\u7d4c\u8def\u306b\u5b58\u5728\u3059\u308b\u30db\u30e2\u30bb\u30ea\u30f3\u8131\u6c34\u7d20\u9175\u7d20\uff08HseDH\uff09\u306f\u3001\u30d2\u30c8\u306b\u306f\u5b58\u5728\u3057\u306a\u3044\u305f\u3081\u3001\u65b0\u898f\u6297\u83cc\u5264\u8a2d\u8a08\u306e\u305f\u3081\u306e\u5206\u5b50\u6a19\u7684\u3068\u3057\u3066\u6ce8\u76ee\u3055\u308c\u308b\u3002\u6211\u3005\u306f\u3001\u8d85\u597d\u71b1\u30a2\u30fc\u30ad\u30a2\u306b\u898b\u51fa\u3057\u305fHseDH\u306b\u3064\u3044\u3066\u69cb\u9020\u89e3\u6790\u3068\u90e8\u4f4d\u7279\u7570\u7684\u5909\u7570\u5b9f\u9a13\u3092\u884c\u3044\u3001NADP\u306e\u5f37\u529b\u306a\u7d50\u5408\u304cNAD(P)\u4f9d\u5b58\u6027\u8131\u6c34\u7d20\u9175\u7d20\u306e\u89e6\u5a92\u6d3b\u6027\u306b\u3068\u3063\u3066\u59a8\u3052\u3068\u306a\u308b\u3068\u3044\u3046\u3001\u88dc\u9175\u7d20\u7279\u7570\u6027\u6c7a\u5b9a\u306e\u65b0\u3057\u3044\u30e1\u30ab\u30cb\u30ba\u30e0\u3092\u660e\u3089\u304b\u306b\u3057\u305f\u3002\u3055\u3089\u306b\u6211\u3005\u306f\u3001\u8d85\u597d\u71b1\u83cc\u7531\u6765 sn<\/em>-\u30b0\u30ea\u30bb\u30ed\u30fc\u30eb-1-\u30ea\u30f3\u9178\u8131\u6c34\u7d20\u9175\u7d20\u306b\u3082\u65b0\u305f\u306a\u88dc\u9175\u7d20\u7d50\u5408\u69d8\u5f0f\u3092\u898b\u51fa\u3057\u3066\u304a\u308a\u3001\u8131\u6c34\u7d20\u9175\u7d20\u306eNAD\/NADP\u7279\u7570\u6027\u3092\u6c7a\u5b9a\u3059\u308b\u30e1\u30ab\u30cb\u30ba\u30e0\u306e\u591a\u69d8\u6027\u3092\u63d0\u5531\u3057\u3066\u3044\u308b\u3002\u3000 \uff08cover image \u21d2\uff09<\/p>\n<\/div>\n\n\n\n

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\u4ee3\u8868\u7684\u306a\u7814\u7a76\u696d\u7e3e<\/strong><\/p>\n<\/div>

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  • Hayashi, J., et al<\/em>. (2017) Structure-based engineering of an artificially generated NADP-dependent D-amino acid  dehydrogenase. Applied and Environmental Microbiology<\/em><\/strong>, in press.<\/li>\n\n\n\n
  • Hayashi, J., et al<\/em>. (2017) Crystal structure of the novel amino-acid racemase isoleucine 2-epimerase from Lactobacillus<\/em> buchneri<\/em>. Acta Crystallographica<\/em><\/strong>, D73, 428-437 .<\/li>\n\n\n\n
  • Wakamatsu, T., et al<\/em>. (2016) Structural insights into L-tryptophan dehydrogenase from a photoautotrophic cyanobacterium Nostocpunctiforme. <\/em>Applied and Environmental Microbiology<\/em>, <\/strong>in press.<\/li>\n\n\n\n
  • Hayashi, J., et al<\/em>. (2016) Unique coenzyme binding mode of hyperthermophilic archaeal sn<\/em>-glycerol-1-phosphate dehydrogenase from Pyrobaculumcalidifontis<\/em>. Proteins<\/em>, <\/strong>84, 1786-1796 .\uff08\u672c\u7814\u7a76\u306fcover image \u306b\u63a1\u629e\u3055\u308c\u305f\uff09<\/li>\n\n\n\n
  • Ohshida, T., et al<\/em>. (2016) First characterization of extremely halophilic 2-deoxy-D-ribose-5-phosphate aldolase.Protein Expression and Purification<\/em><\/strong>, <\/em>126, 62-68 .<\/li>\n\n\n\n
  • Hayashi, J., et al<\/em>. (2015) Crystal structures of a hyperthermophilic archaeal homoserine dehydrogenase suggest a novel cofactor binding mode for oxidoreductases. Scientific Reports<\/em><\/strong>, 5, 11674.<\/li>\n\n\n\n
  • Sakuraba, H., et al<\/em>. (2012) Crystal structure of novel dye-linked L-proline dehydrogenase from hyperthermophilic  archaeon Aeropyrum pernix. <\/em>The<\/em><\/strong> <\/em>Journal of Biological Chemistry<\/em>, <\/strong>287, 20070-20080.<\/li>\n<\/ul>\n\n<\/div>
    \n\n
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    Research Area: Life Science & Biotechnology<\/mark> \u3000\u3000
    Research Specialization :\u3000Enzyme Engineering<\/mark> \u3000
    Name:\u3000SAKURABA, Haruhiko<\/mark><\/h2>\n\n\n\n
    \"\"<\/figure>\n\n\n\n

    Keywords: hyperthermophiles, archaea, thermostable enzyme,x-ray crystal structure analysis<\/p>\n<\/div>\n\n\n\n

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    Recent Research<\/strong><\/strong><\/h2>\n\n\n\n

    1. Structure-based creation of a novel D-amino acid dehydrogenase<\/strong><\/strong><\/h2>\n\n\n\n

       meso<\/em>-Diaminopimelate dehydrogenase (DAPDH) acts stereoselectively on the D-center of\u3000 substrate. Thus, the enzyme is expected to be useful for one-step production of D-amino acids, which are often utilized as source materials for industrial production of medicines and agrochemicals. However, its high substrate specificity for DAP and instability have proved to be the major disadvantage for the practical application of DAPDH. Recently, we determined the crystal structure of thermostable DAPDH and estimated the amino acid residues related to substrate binding. We used site-directed mutagenesis to identify the residues responsible for determining the enzyme\u2019s substrate specificity and found that a single mutation (D94A) caused a striking change in the substrate specificity of the enzyme. The mutant exhibited extremely high activity for reductive amination of phenylpyruvate. This is a major advantage of using the D94A mutant for one-step synthesis of D-phenylalanine.<\/p>\n\n\n\n

    2. Unique coenzyme binding mode of hyperthermophilicdehydrogenase<\/strong><\/strong><\/h2>\n\n\n\n
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    \u3000 Homoserine dehydrogenase (HseDH) is a key enzyme in the biosynthetic pathway for the synthesis of methionine, threonine, and isoleucine in plants and microorganisms. HseDH is thought to be a potential target for the structure-based design of antibiotics, as the enzyme is not present in mammals. We recently determined the crystal structure of HseDH from the hyperthermophile and found that this enzyme exhibits a new variation on cofactor binding to a dehydrogenase: very strong NADP binding that acts as an obstacle to NAD(P)-dependent dehydrogenase catalytic activity. More recently, we observed another variation on a structural basis for cofactor preference using hyper-thermophilic sn<\/em>-glycerol-1-phosphate dehydrogenase. Our studies indicate that the molecular basis for the cofactor preference in NAD(P)-dependent dehydrogenases may have much more variation than expected<\/p>\n<\/div>\n\n\n\n

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    \u4ee3\u8868\u7684\u306a\u7814\u7a76\u696d\u7e3e<\/strong><\/p>\n<\/div>

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      \n
    • Hayashi, J., et al<\/em>. (2017) Structure-based engineering of an artificially generated NADP-dependent D-amino acid  dehydrogenase. Applied and Environmental Microbiology<\/em><\/strong>, in press.<\/li>\n\n\n\n
    • Hayashi, J., et al<\/em>. (2017) Crystal structure of the novel amino-acid racemase isoleucine 2-epimerase from Lactobacillus<\/em> buchneri<\/em>. Acta Crystallographica<\/em><\/strong>, D73, 428-437 .<\/li>\n\n\n\n
    • Wakamatsu, T., et al<\/em>. (2016) Structural insights into L-tryptophan dehydrogenase from a photoautotrophic cyanobacterium Nostocpunctiforme. <\/em>Applied and Environmental Microbiology<\/em>, <\/strong>in press.<\/li>\n\n\n\n
    • Hayashi, J., et al<\/em>. (2016) Unique coenzyme binding mode of hyperthermophilic archaeal sn<\/em>-glycerol-1-phosphate dehydrogenase from Pyrobaculumcalidifontis<\/em>. Proteins<\/em>, <\/strong>84, 1786-1796 .\uff08\u672c\u7814\u7a76\u306fcover image \u306b\u63a1\u629e\u3055\u308c\u305f\uff09<\/li>\n\n\n\n
    • Ohshida, T., et al<\/em>. (2016) First characterization of extremely halophilic 2-deoxy-D-ribose-5-phosphate aldolase.Protein Expression and Purification<\/em><\/strong>, <\/em>126, 62-68 .<\/li>\n\n\n\n
    • Hayashi, J., et al<\/em>. (2015) Crystal structures of a hyperthermophilic archaeal homoserine dehydrogenase suggest a novel cofactor binding mode for oxidoreductases. Scientific Reports<\/em><\/strong>, 5, 11674.<\/li>\n\n\n\n
    • Sakuraba, H., et al<\/em>. (2012) Crystal structure of novel dye-linked L-proline dehydrogenase from hyperthermophilic  archaeon Aeropyrum pernix. <\/em>The<\/em><\/strong> <\/em>Journal of Biological Chemistry<\/em>, <\/strong>287, 20070-20080.<\/li>\n<\/ul>\n\n<\/div><\/div>\n <\/div>","protected":false},"excerpt":{"rendered":"","protected":false},"author":1,"featured_media":15781,"menu_order":0,"comment_status":"open","ping_status":"open","template":"","meta":{"_coblocks_attr":"","_coblocks_dimensions":"","_coblocks_responsive_height":"","_coblocks_accordion_ie_support":"","ub_ctt_via":"","footnotes":""},"class_list":["post-3131","research","type-research","status-publish","has-post-thumbnail","hentry","research_keyword-202","research_keyword-201","research_keyword-200","research_keyword-203","research_category-184","center_category-258","center_category-260","center_category-259","sdgs_category-12"],"_links":{"self":[{"href":"https:\/\/www.ag.kagawa-u.ac.jp\/index.php?rest_route=\/wp\/v2\/research\/3131","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.ag.kagawa-u.ac.jp\/index.php?rest_route=\/wp\/v2\/research"}],"about":[{"href":"https:\/\/www.ag.kagawa-u.ac.jp\/index.php?rest_route=\/wp\/v2\/types\/research"}],"author":[{"embeddable":true,"href":"https:\/\/www.ag.kagawa-u.ac.jp\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.ag.kagawa-u.ac.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=3131"}],"version-history":[{"count":72,"href":"https:\/\/www.ag.kagawa-u.ac.jp\/index.php?rest_route=\/wp\/v2\/research\/3131\/revisions"}],"predecessor-version":[{"id":32741,"href":"https:\/\/www.ag.kagawa-u.ac.jp\/index.php?rest_route=\/wp\/v2\/research\/3131\/revisions\/32741"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.ag.kagawa-u.ac.jp\/index.php?rest_route=\/wp\/v2\/media\/15781"}],"wp:attachment":[{"href":"https:\/\/www.ag.kagawa-u.ac.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=3131"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}