{"id":44657,"date":"2023-02-22T08:00:07","date_gmt":"2023-02-22T00:00:07","guid":{"rendered":"\/\/www.gsbet888.com\/?p=44657"},"modified":"2023-02-21T21:15:35","modified_gmt":"2023-02-21T13:15:35","slug":"%e4%b8%9b%e5%bf%97%e5%a5%87%e7%a0%94%e7%a9%b6%e5%91%98%e5%9b%a2%e9%98%9fjacs%ef%bc%9a%e9%9a%a7%e9%81%93%e5%b7%a5%e7%a8%8b%e7%90%86%e6%80%a7%e8%ae%be%e8%ae%a1p450%e8%bf%87%e5%8a%a0%e6%b0%a7%e9%85%b6","status":"publish","type":"post","link":"\/\/www.gsbet888.com\/%e5%8c%96%e5%ad%a6%e6%9d%82%e8%ae%b0\/recentpaper\/2023\/02\/%e4%b8%9b%e5%bf%97%e5%a5%87%e7%a0%94%e7%a9%b6%e5%91%98%e5%9b%a2%e9%98%9fjacs%ef%bc%9a%e9%9a%a7%e9%81%93%e5%b7%a5%e7%a8%8b%e7%90%86%e6%80%a7%e8%ae%be%e8%ae%a1p450%e8%bf%87%e5%8a%a0%e6%b0%a7%e9%85%b6.html","title":{"rendered":"\u4e1b\u5fd7\u5947\u7814\u7a76\u5458\u56e2\u961fJACS\uff1a\u96a7\u9053\u5de5\u7a0b\u7406\u6027\u8bbe\u8ba1P450\u8fc7\u52a0\u6c27\u9176\u7684\u65b0\u7b56\u7565"},"content":{"rendered":"<p>\u4f5c\u8005\uff1a\u77f3\u6cb9\u919a<\/p>\n<h4><strong><b>\u5bfc\u8bed\uff1a<\/b><\/strong><\/h4>\n<p>\u8fd1\u65e5\uff0c<strong><b>\u4e2d\u56fd\u79d1\u5b66\u9662\u9752\u5c9b\u751f\u7269\u80fd\u6e90\u4e0e\u8fc7\u7a0b\u7814\u7a76\u6240\u4e1b\u5fd7\u5947\u7814\u7a76\u5458\u56e2\u961f<\/b><\/strong>\u901a\u8fc7\u7406\u6027\u8bbe\u8ba1P450\u9176\u7684\u6c34\u5206\u5b50\u96a7\u9053\uff08water tunnel\uff09\u6765\u4fc3\u8fdb\u8fc7\u6c27\u5316\u6c22\uff08H<sub>2<\/sub>O<sub>2<\/sub>\uff09\u8fdb\u5165\u5176\u5185\u90e8\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\uff0c\u6210\u529f\u5c06\u4e24\u79cdNADH\u4f9d\u8d56\u578bP450\u5355\u52a0\u6c27\u9176\uff08CYP199A4\u548cCYP153A<em><sub>M.aq<\/sub><\/em>\uff09\u6539\u9020\u4e3a\u5177\u6709\u9ad8\u50ac\u5316\u6d3b\u6027\u7684P450\u8fc7\u52a0\u6c27\u9176\uff0c\u5e76\u4e14\u6210\u529f\u5e94\u7528\u5230\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u534f\u540cP450BM3\u8fc7\u52a0\u6c27\u9176\u4f53\u7cfb\u3002\u96a7\u9053\u5de5\u7a0b\u6709\u671b\u6210\u4e3a\u4e00\u4e2a\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u8fc7\u52a0\u6c27\u9176\u7684\u666e\u9002\u6027\u65b0\u7b56\u7565\uff0c\u8be5\u6210\u679c\u4e8e\u8fd1\u671f\u53d1\u8868\u4e8e<em><i>J. Am. Chem. Soc<\/i><\/em><em><i>.<\/i><\/em>\u4e0a\u3002<\/p>\n<blockquote><p>\u201cEnabling Peroxygenase Activity in Cytochrome P450 Monooxygenases by Engineering Hydrogen Peroxide Tunnel.<\/p>\n<p>Panxia Zhao,<sup>\u2020<\/sup>\u00a0Fanhui Kong,<sup>\u2020<\/sup>\u00a0Yiping Jiang,<sup>\u2020<\/sup>\u00a0Xiangquan Qin, Xiaoxia Tian, and Zhiqi Cong<sup>*<\/sup><em><i>.<\/i><\/em><\/p>\n<p><em><i>J. Am. Chem. Soc.<\/i><\/em><strong><b>2023<\/b><\/strong><strong><b>,<\/b><\/strong><strong><b>ASAP<\/b><\/strong>.DOI <a href=\"https:\/\/pubs.acs.org\/doi\/10.1021\/jacs.3c00195\"><u>10.1021\/jacs.3c00195<\/u><\/a>.\u201d<\/p><\/blockquote>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-44659 size-full\" src=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/543a5096b6465cabc00c10cb9a6883de.png\" alt=\"\" width=\"517\" height=\"421\" srcset=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/543a5096b6465cabc00c10cb9a6883de.png 517w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/543a5096b6465cabc00c10cb9a6883de-300x244.png 300w\" sizes=\"(max-width: 517px) 100vw, 517px\" \/><\/p>\n<h4><strong><b>\u6b63\u6587\uff1a<\/b><\/strong><\/h4>\n<p>\u7ec6\u80de\u8272\u7d20P450\u5355\u52a0\u6c27\u9176\u662f\u81ea\u7136\u754c\u4e2d\u6700\u91cd\u8981\u7684\u591a\u529f\u80fd\u751f\u7269\u6c27\u5316\u50ac\u5316\u5242\u4e4b\u4e00\uff0c\u5e7f\u6cdb\u53c2\u4e0e\u836f\u7269\u4ee3\u8c22\u3001\u5929\u7136\u4ea7\u7269\u5408\u6210\u3001\u5f02\u6e90\u7269\u8d28\u964d\u89e3\u548c\u753e\u4f53\u6fc0\u7d20\u7684\u751f\u7269\u5408\u6210\uff0c\u5e76\u4e14\u5176\u5bf9\u6781\u5177\u6311\u6218\u6027\u7684\u60f0\u6027\u78b3-\u6c22\u952e\u9009\u62e9\u6c27\u5316\u53cd\u5e94\u5177\u6709\u4f18\u5f02\u7684\u50ac\u5316\u80fd\u529b\u3002\u5927\u591a\u6570P450\u5355\u52a0\u6c27\u9176\u8981\u901a\u8fc7\u590d\u6742\u7684\u6c27\u5316\u8fd8\u539f\u4f53\u7cfb\u5b9e\u73b0\u5176\u50ac\u5316\u529f\u80fd\uff0c\u5bfc\u81f4\u5176\u5b9e\u9645\u5e94\u7528\u53d7\u5230\u4e00\u5b9a\u7684\u9650\u5236\u3002\u5229\u7528H<sub>2<\/sub>O<sub>2<\/sub>\u4ee3\u66ffO<sub>2<\/sub>\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u8fc7\u52a0\u6c27\u9176\uff0c\u88ab\u8ba4\u4e3a\u662f\u5f00\u53d1\u7b80\u5355\u5b9e\u7528\u578bP450\u751f\u7269\u50ac\u5316\u5242\u7684\u6709\u6548\u9014\u5f84\uff0c\u4f46\u662f\u7edd\u5927\u591a\u6570P450\u5355\u52a0\u6c27\u9176\u5229\u7528H<sub>2<\/sub>O<sub>2<\/sub>\u8fdb\u884c\u50ac\u5316\u53cd\u5e94\u4ec5\u6709\u6781\u5f31\u3001\u751a\u81f3\u6ca1\u6709\u50ac\u5316\u6d3b\u6027\u3002<\/p>\n<p>\u8fd1\u5e74\u6765\uff0c\u9752\u5c9b\u80fd\u6e90\u6240\u4e1b\u5fd7\u5947\u7814\u7a76\u5458\u56e2\u961f\u81f4\u529b\u4e8eP450\u8fc7\u52a0\u6c27\u9176\u7684\u5206\u5b50\u8bbe\u8ba1\u548c\u521b\u5236\uff0c\u5728\u56fd\u9645\u4e0a\u9996\u6b21\u63d0\u51fa\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u534f\u540cP450\u9176\u50ac\u5316\u6982\u5ff5\uff0c\u6210\u529f\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u80fd\u9ad8\u6548\u5229\u7528H<sub>2<\/sub>O<sub>2<\/sub>\u7684P450\u8fc7\u52a0\u6c27\u9176\uff08<em>Angew. Chem. Int. Ed<\/em>. <strong><b>2018<\/b><\/strong>,\u00a07628 Very Important Paper\uff09\uff0c\u5e76\u901a\u8fc7\u5408\u4f5c\u9610\u660e\u4e86\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u534f\u540cP450\u8fc7\u52a0\u6c27\u9176\u7684\u7ed3\u6784\u57fa\u7840\u548c\u50ac\u5316\u673a\u7406\uff08<em>ACS Catal.<\/em><em>\u00a0<\/em><strong>2021<\/strong>, 8774\uff09\u3002\u7814\u7a76\u4eba\u5458\u8fdb\u800c\u7ed3\u5408\u86cb\u767d\u8d28\u5de5\u7a0b\u6280\u672f\uff0c\u5148\u540e\u5f00\u53d1\u51fa\u4e86H<sub>2<\/sub>O<sub>2<\/sub>\u9a71\u52a8\u7684P450\u4e19\u70f7\u7f9f\u5316\u9176\uff08<em><i>ACS Catal. <\/i><\/em><strong><b>2019<\/b><\/strong>, 7350\uff09\u3001\u9ad8\u5ea6<em><i>R<\/i><\/em>-\u5bf9\u6620\u9009\u62e9\u6027\u7684P450\u82ef\u4e59\u70ef\u73af\u6c27\u5316\u9176\uff08<em><i>Chem. Sci<\/i><\/em>. <strong><b>2021<\/b><\/strong>, 6307\uff09\u3001\u9ad8\u6548P450\u8fc7\u6c27\u5316\u7269\u9176\uff08<em>ACS Catal.<\/em><em>\u00a0<\/em><strong>2021<\/strong>, 8449\uff09\u3001\u78b3-\u6c22\u952e\u7684\u533a\u57df\u548c\u7acb\u4f53\u591a\u6837\u6027\u7f9f\u5316P450\u5de5\u7a0b\u9176\uff08<em>Angew. Chem. Int. Ed<\/em>. <strong><b>20<\/b><\/strong><strong><b>23<\/b><\/strong>, e202215088. Hot\u00a0Paper\uff09\u3001\u4e0d\u9971\u548c\u70c3\u76f4\u63a5\u785d\u5316\u7684P450\u785d\u5316\u9176\uff08<em>Angew. Chem. Int. Ed<\/em>. <strong><b>2023<\/b><\/strong>, e202217678.\uff09\u3002\u4f46\u662f\u7814\u7a76\u4eba\u5458\u53d1\u73b0\u9700\u8981\u9ad8\u51fa\u5e95\u7269\u6d53\u5ea6\u6570\u500d\u7684H<sub>2<\/sub>O<sub>2<\/sub>\u624d\u80fd\u5b9e\u73b0\u9ad8\u50ac\u5316\u6d3b\u6027\uff0c\u8fd9\u5bf9\u66f4\u5177\u5b9e\u7528\u6027P450\u9176\u50ac\u5316\u5242\u7684\u5f00\u53d1\u63d0\u51fa\u4e86\u65b0\u7684\u6311\u6218\u3002<\/p>\n<p>\u53e6\u5916\uff0c\u7814\u7a76\u4eba\u5458\u5229\u7528\u5b9a\u70b9\u7a81\u53d8\u3001\u5b9a\u5411\u8fdb\u5316\u548c\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u7b56\u7565\uff08\u56fe1\uff09\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u8fc7\u52a0\u6c27\u9176\uff0c\u5c06\u8fc7\u52a0\u6c27\u9176\u4e0e\u5929\u7136\u8fc7\u6c27\u5316\u7269\u9176\u7684\u7ed3\u6784\u6bd4\u5bf9\u5206\u6790\u53d1\u73b0\uff1a\u5929\u7136\u8fc7\u6c27\u5316\u7269\u9176\u7684\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\u66f4\u591a\u7684\u66b4\u9732\u4e8e\u86cb\u767d\u8868\u9762\uff0c\u6216\u8005\u6709\u8f83\u4e3a\u5f00\u653e\u7684\u5e95\u7269\u901a\u9053\u4e0e\u86cb\u767d\u8868\u9762\u76f8\u901a\uff0c\u6240\u4ee5H<sub>2<\/sub>O<sub>2<\/sub>\u5206\u5b50\u80fd\u591f\u66f4\u5bb9\u6613\u5230\u8fbe\u6d3b\u6027\u4e2d\u5fc3\u3002\u76f8\u53cd\uff0c\u70df\u9170\u80fa\u8f85\u56e0\u5b50NAD(P)H\u4f9d\u8d56\u578bP450\u9176\u7684\u6d3b\u6027\u4e2d\u5fc3\u5927\u90fd\u6df1\u57cb\u5728\u86cb\u767d\u5185\u90e8\uff0cH<sub>2<\/sub>O<sub>2<\/sub>\u5206\u5b50\u53ef\u80fd\u4e0d\u5bb9\u6613\u901a\u8fc7\u758f\u6c34\u6027\u5e95\u7269\u901a\u9053\u8fdb\u5165\u6d3b\u6027\u4e2d\u5fc3\uff0c\u5bfc\u81f4\u5de5\u7a0b\u5316P450\u8fc7\u52a0\u6c27\u9176\u5728\u8fdb\u884c\u50ac\u5316\u53cd\u5e94\u65f6\u901a\u5e38\u9700\u8981\u9ad8\u6d53\u5ea6\u7684H<sub>2<\/sub>O<sub>2<\/sub>\u3002<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-44660 size-full\" src=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/130c135258708adde0afd21f603bc072.png\" alt=\"\" width=\"650\" height=\"458\" srcset=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/130c135258708adde0afd21f603bc072.png 650w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/130c135258708adde0afd21f603bc072-300x211.png 300w\" sizes=\"(max-width: 650px) 100vw, 650px\" \/><\/p>\n<p>Figure 1. Schematic diagram of NAD(P)H-dependent P450 monooxygenases and H<sub>2<\/sub>O<sub>2<\/sub>-driven P450 peroxygenases and strategies for modifying P450 monooxygenase into its peroxygenase mode.<\/p>\n<p>\u9274\u4e8eP450\u9176\u7ed3\u6784\u4e2d\u5b58\u5728\u8fde\u63a5\u86cb\u767d\u8868\u9762\u4e0e\u6d3b\u6027\u4e2d\u5fc3\u7684H<sub>2<\/sub>O\u5206\u5b50\u96a7\u9053\uff08water tunnel\uff09\uff0c\u7814\u7a76\u4eba\u5458\u63d0\u51fa\u901a\u8fc7P450\u9176\u7684\u6c34\u96a7\u9053\u8bbe\u8ba1\u6765\u4fc3\u8fdbH<sub>2<\/sub>O<sub>2<\/sub>\u8fdb\u5165\u5176\u5185\u90e8\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\uff0c\u589e\u5f3aP450\u8fc7\u52a0\u6c27\u9176\u7684\u50ac\u5316\u6d3b\u6027\u3002\u7814\u7a76\u4eba\u5458\u5229\u7528Caver web 1.0\u9884\u6d4b\u51faP450\u9176\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\u4e0e\u6eb6\u5242\u73af\u5883\u4e4b\u95f4\u7684\u6c34\u96a7\u9053\uff0c\u901a\u8fc7\u8ba1\u7b97\u5206\u6790\u9274\u5b9a\u51fa\u53ef\u80fd\u5f71\u54cdH<sub>2<\/sub>O<sub>2<\/sub>\u6d41\u5165\u6d3b\u6027\u4e2d\u5fc3\u7684\u96a7\u9053\u5165\u53e3\u6216\u74f6\u9888\u5904\u7684\u5173\u952e\u6c28\u57fa\u9178\uff0c\u7ed3\u5408\u5b9a\u70b9\u7a81\u53d8\u548c\u591a\u8f6e\u53cd\u5e94\u7b5b\u9009\uff0c\u6210\u529f\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u5de5\u7a0b\u5316\u7684P450\u8fc7\u52a0\u6c27\u9176\u3002<\/p>\n<p>\u9996\u5148\uff0c\u7814\u7a76\u4eba\u5458\u5229\u7528\u96a7\u9053\u5de5\u7a0b\u5bf9\u6765\u81ea<em>Rhodopseudomonas palustris <\/em>HaA2\u00a0NADH\u4f9d\u8d56\u7684CYP199A4\u8fdb\u884c\u7814\u7a76\u3002\u7ed3\u679c\u8868\u660eH<sub>2<\/sub>O\u5206\u5b50\u96a7\u9053\u74f6\u9888\u6216\u5165\u53e3\u7684\u5173\u952e\u6c28\u57fa\u9178P90\u3001F298\u548cF182\u7684\u7a81\u53d8\u6709\u8f83\u4f18\u50ac\u5316\u6027\u80fd\uff08\u56fe2A\uff09\uff0cF182A\u50ac\u53164-\u7532\u6c27\u57fa\u82ef\u7532\u9178<em>O<\/em>-\u8131\u7532\u57fa\u5316\u6d3b\u6027\u76f8\u6bd4\u91ce\u751f\u578b\u63d0\u5347\u4e86163\u500d\uff08\u56fe2B\uff09\uff0c\u5176\u50ac\u5316\u8f6c\u5316\u6570\uff08TON\uff09\u4e3a980\u00b5M(\u00b5M P450)<sup>-1<\/sup>\u3002\u968f\u540e\uff0c\u7814\u7a76\u4eba\u5458\u5c06\u6709\u76ca\u7a81\u53d8\u4f53\u8fdb\u884c\u7ec4\u5408\u7a81\u53d8\uff0c\u53d1\u73b0\u53cc\u7a81\u53d8F182A\/P90A\u7684TON\u8fbe\u52301100\u00b5M(\u00b5M P450)<sup>-1<\/sup>(\u589e\u52a0183\u500d)\uff08\u56fe2B\uff09\u3002<\/p>\n<p>\u5728\u4e0a\u8ff0\u57fa\u7840\u4e0a\uff0c\u7814\u7a76\u4eba\u5458\u4e3a\u4e86\u8fdb\u4e00\u6b65\u9a8c\u8bc1\u8be5\u7b56\u7565\u7684\u666e\u9002\u6027\uff0c\u5c06\u4e0a\u8ff0\u7b56\u7565\u5e94\u7528\u4e8e\u6765\u81ea<em>Marinobacter aquaeolei<\/em>\u00a0\u7684NADH\u4f9d\u8d56\u7684CYP153A<em><sub>M.aq<\/sub><\/em>\u3002\u7ed3\u679c\u8868\u660e\uff1a\u4e0e\u91ce\u751f\u578b\u76f8\u6bd4\uff0c\u76f8\u540c\u96a7\u9053\u5185\u7684\u5173\u952e\u6c28\u57fa\u9178D226\u3001D310\u548cV456\u50ac\u5316\u6708\u6842\u9178\u7684\u7f9f\u5316\u6d3b\u6027\u63d0\u5347\u81f313\u500d\u4ee5\u4e0a\uff08\u56fe2D-2E\uff09\uff0c\u5c06\u4e0a\u8ff0\u5173\u952e\u6c28\u57fa\u9178\u8fdb\u884c\u7ec4\u5408\u7a81\u53d8\uff0c\u53d1\u73b0\u53cc\u7a81\u53d8\u4f53D266A\/V456A\u548cD266T\/V456G\u7684TON\u5206\u522b\u63d0\u9ad8\u81f3161\u00b5M\u548c167\u00b5M(\u00b5M P450)<sup>-1<\/sup>\uff0c\u6bd4WT\u63d0\u9ad815\u500d\uff08\u56fe2E\uff09\u3002<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-44661 size-full\" src=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/ebe19a97f50fc839e68b131d154b011b.png\" alt=\"\" width=\"499\" height=\"496\" srcset=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/ebe19a97f50fc839e68b131d154b011b.png 499w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/ebe19a97f50fc839e68b131d154b011b-300x298.png 300w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/ebe19a97f50fc839e68b131d154b011b-150x150.png 150w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/ebe19a97f50fc839e68b131d154b011b-120x120.png 120w\" sizes=\"(max-width: 499px) 100vw, 499px\" \/> <img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-44662 size-full\" src=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/c9387e5d1fb837b85607535f5fabf4c8.png\" alt=\"\" width=\"491\" height=\"495\" srcset=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/c9387e5d1fb837b85607535f5fabf4c8.png 491w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/c9387e5d1fb837b85607535f5fabf4c8-298x300.png 298w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/c9387e5d1fb837b85607535f5fabf4c8-150x150.png 150w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/c9387e5d1fb837b85607535f5fabf4c8-120x120.png 120w\" sizes=\"(max-width: 491px) 100vw, 491px\" \/><\/p>\n<p style=\"text-align: left;\">Figure 2. Engineering the peroxygenase activity of CYP199A4 and CYP153A<em><sub><i>M.aq<\/i><\/sub><\/em>.<\/p>\n<p>\u540e\u7eed\uff0c\u4e3a\u4e86\u63a2\u7a76\u96a7\u9053\u5de5\u7a0b\u63d0\u9ad8P450\u8fc7\u52a0\u6c27\u9176\u6d3b\u6027\u7684\u50ac\u5316\u673a\u5236\uff0c\u7814\u7a76\u4eba\u5458\u5206\u522b\u89e3\u6790\u4e86\u9ad8\u5206\u8fa8\u7387\u7684CYP199A4 F182T\u00a0\u548c\u4e0e4-\u7532\u6c27\u57fa\u82ef\u7532\u9178\u7684\u6676\u4f53\u7ed3\u6784\uff08\u56fe3A\uff09\u3002\u4e0e\u65e9\u671f\u7814\u7a76\u7684CYP199A4\u7a81\u53d8\u4f53\u5982244D\u3001D251N\u548cT252A (PDB: 7N60\u30015KYD\u548c5KDZ)\u76f8\u6bd4\uff0c\u5728CYP199A4 F182T\u6d3b\u6027\u53e3\u888b\u5185\u89c2\u5bdf\u5230\u56db\u4e2a\u65b0\u7684\u6c34\u5206\u5b50(w1-w4)\uff0c\u5e76\u4e0eF182T\u5468\u56f4\u7684\u6c34\u5206\u5b50(w5-w8)\u5f62\u6210\u4e00\u6761\u901a\u5f80\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\u7684\u96a7\u9053\uff0c\u5e76\u4e14F182T\u5bfc\u81f4D251\u4fa7\u94fe\u5411\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\u53d1\u751f\u7ffb\u8f6c\uff0c\u4e0e\u6d3b\u6027\u4e2d\u5fc3\u7684\u6c34\u5206\u5b50\u5f62\u6210\u6c22\u952e\u7f51\u7edc\uff0c\u4fc3\u8fdbH<sub>2<\/sub>O<sub>2<\/sub>\u7684\u6d3b\u5316\u3002\u53e6\u5916\uff0c\u5bf9\u4e8eCYP153A<em><sub>M.aq <\/sub><\/em>V456A\u4e0e\u6708\u6842\u9178\u7684\u6676\u4f53\u7ed3\u6784\uff0c\u5176\u5bf9\u5e94\u6c34\u96a7\u9053\u7684\u6c34\u5206\u5b50\u6bd4WT\u591a\u4e865\u4e2a\uff0c\u8fdb\u800c\u518d\u6b21\u8bc1\u660e\u4e86\u6c34\u96a7\u9053\u5de5\u7a0b\u7b56\u7565\u7684\u786e\u6709\u5229\u4e8e\u6c34\u6216\u8fc7H<sub>2<\/sub>O<sub>2<\/sub>\u8fdb\u5165\u6d3b\u6027\u4e2d\u5fc3\uff0c\u6709\u529b\u652f\u6301\u4e86\u6700\u521d\u7684\u5047\u8bbe\u3002<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-44663 size-full\" src=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/568ac2dec6da68bd59b9184842e9ab02.png\" alt=\"\" width=\"494\" height=\"395\" srcset=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/568ac2dec6da68bd59b9184842e9ab02.png 494w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/568ac2dec6da68bd59b9184842e9ab02-300x240.png 300w\" sizes=\"(max-width: 494px) 100vw, 494px\" \/> <img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-44664 size-full\" src=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/6b3326d87e4dedd82a9b7b39b8148d81.png\" alt=\"\" width=\"519\" height=\"409\" srcset=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/6b3326d87e4dedd82a9b7b39b8148d81.png 519w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/6b3326d87e4dedd82a9b7b39b8148d81-300x236.png 300w\" sizes=\"(max-width: 519px) 100vw, 519px\" \/><\/p>\n<p style=\"text-align: left;\">Figure 3. (A) Active site of CYP199A4 F182T and\u00a04-MBA (PDB: 8HGC), where hydrogen bonds are drawn as black dashed lines; (B) Active site of CYP153A<em><sub><i>M.aq<\/i><\/sub><\/em>\u00a0V456A (PDB: 8HGT), where the substrate binding site is indicated with a yellow stick model of the product molecule based on a solved structure containing the product 12-HDA (PDB: 5FYG).<\/p>\n<p>\u6700\u540e\uff0c\u7814\u7a76\u4eba\u5458\u5c06\u96a7\u9053\u5de5\u7a0b\u7b56\u7565\u4e0e\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u534f\u540cP450BM3\u8fc7\u52a0\u6c27\u9176\u4f53\u7cfb\u76f8\u7ed3\u5408\uff0c\u9009\u7528\u5432\u54da\u4e3a\u6a21\u5f0f\u53cd\u5e94\uff0c\u4ee5F87G\/T268V (GV) \u4e3a\u7236\u672c\u9176\uff0c\u7b5b\u9009\u83b7\u5f97\u5173\u952e\u6c28\u57fa\u9178D182G\u548cL272S(\u56fe4A)\u3002\u7ed3\u679c\u663e\u793a\uff1a\u76f8\u6bd4GV\uff0cGV\/D182G\u548cGV\/L272S\u7684\u50ac\u5316\u6d3b\u6027\u5206\u522b\u63d0\u9ad8\u4e86114%\u548c153%\uff0c\u4e24\u4e2a\u4f18\u52bf\u7a81\u53d8\u4f53\u540c\u6837\u5bf9\u82ef\u4e59\u70ef\u73af\u6c27\u5316\u3001\u4e59\u82ef\u7f9f\u5316\u548c\u82ef\u7532\u786b\u919a\u4e9a\u78fa\u5316\u5e95\u7684\u50ac\u5316\u6d3b\u6027\u663e\u8457\u63d0\u9ad8\uff08\u56fe4C\uff09\u3002\u5176\u4e2dGV\/L272S\u5c06H<sub>2<\/sub>O<sub>2<\/sub>\u6d53\u5ea6\u964d\u4f4e\u81f3\u63a5\u8fd1\u5929\u7136\u8fc7\u52a0\u6c27\u9176\u6c34\u5e73\uff0c\u5176\u50ac\u5316\u5432\u54da\u751f\u6210\u4ea7\u7269\u6240\u9700H<sub>2<\/sub>O<sub>2<\/sub>\u6d53\u5ea6\u88ab\u964d\u4f4e\u4e8695%\u4ee5\u4e0a\uff0cH<sub>2<\/sub>O<sub>2<\/sub>\u4e0e\u4ea7\u7269\u7684\u8026\u5408\u6548\u7387\u8fbe\u523095%\u3002\u6b64\u5916\uff0c\u7814\u7a76\u4eba\u5458\u5229\u7528Caver\u5206\u6790\u4e8621B3\u7684\u7236\u672c\u9176P450BM3 F87A\u7684\u6c34\u96a7\u9053\uff0c\u7ed3\u679c\u53d1\u73b021B3\u4e2d4\u4e2a\u7a81\u53d8\u4f4d\u70b9\u4f4d\u4e8e\u6c34\u96a7\u9053\u7684\u74f6\u9888\u5904\uff08\u56fe4B\uff09\u3002\u8fdb\u800c\u518d\u6b21\u8bc1\u660e\u4e86\u6c34\u96a7\u9053\u5de5\u7a0b\u7b56\u7565\u53ef\u4fc3\u8fdbP450\u8fc7\u52a0\u6c27\u9176\u6d3b\u6027\u3002<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-44665 size-full\" src=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/344dabd9a368469c9b195fce541be1e1.png\" alt=\"\" width=\"585\" height=\"270\" srcset=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/344dabd9a368469c9b195fce541be1e1.png 585w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/344dabd9a368469c9b195fce541be1e1-300x138.png 300w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/344dabd9a368469c9b195fce541be1e1-370x170.png 370w\" sizes=\"(max-width: 585px) 100vw, 585px\" \/> <img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-44666 size-full\" src=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/9d54bd1d24120f7ac2400da83caa23cb.png\" alt=\"\" width=\"446\" height=\"270\" srcset=\"https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/9d54bd1d24120f7ac2400da83caa23cb.png 446w, https:\/\/assets.www.gsbet888.com\/uploads\/2023\/02\/9d54bd1d24120f7ac2400da83caa23cb-300x182.png 300w\" sizes=\"(max-width: 446px) 100vw, 446px\" \/><\/p>\n<p>Figure 4. Engineered water tunnels improve the catalytic activity of DFSM-facilitated P450BM3 peroxygenase.<\/p>\n<h4><strong><b>\u603b\u7ed3<\/b><\/strong><strong><b>:<\/b><\/strong><\/h4>\n<p>\u672c\u7814\u7a76\u63d0\u51fa\u4e86\u57fa\u4e8e\u96a7\u9053\u5de5\u7a0b\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u8fc7\u52a0\u6c27\u9176\u7684\u65b0\u7b56\u7565\uff0c\u8be5\u7b56\u7565\u6210\u529f\u5e94\u7528\u4e8eNADH\u4f9d\u8d56\u7684\u5929\u7136P450\u5355\u52a0\u6c27\u9176(CYP199A4\u548cCYP153A<em><sub>M.aq<\/sub><\/em>)\u548c\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u4fc3\u8fdb\u7684P450BM3\u8fc7\u52a0\u6c27\u9176\uff0c\u5e76\u4e14\u8fdb\u4e00\u6b65\u589e\u5f3a\u5176\u8fc7\u52a0\u6c27\u9176\u6d3b\u6027\u3002\u6709\u671b\u4e3a\u4eba\u5de5P450\u8fc7\u52a0\u6c27\u9176\u9ad8\u6548\u5229\u7528H<sub>2<\/sub>O<sub>2<\/sub>\u63d0\u4f9b\u4e00\u79cd\u6709\u524d\u666f\u3001\u53ef\u63a8\u5e7f\u7684\u7b56\u7565\uff0c\u8fdb\u4e00\u6b65\u62d3\u5c55\u4e86P450\u5728\u5408\u6210\u5316\u5b66\u548c\u5408\u6210\u751f\u7269\u5b66\u4e2d\u7684\u5e94\u7528\u8303\u56f4\u3002<\/p>\n<p>\u8be5\u5de5\u4f5c\u7531\u9752\u5c9b\u80fd\u6e90\u6240\u535a\u58eb\u751f<strong><b>\u8d75\u76fc\u971e\u3001\u5b54\u7e41\u6167<\/b><\/strong>\u548c<strong><b>\u59dc\u8c0a\u5e73<\/b><\/strong>\u535a\u58eb\u5171\u540c\u5b8c\u6210\uff0c\u4e3a\u8bba\u6587\u5171\u540c\u7b2c\u4e00\u4f5c\u8005\uff0c\u9752\u5c9b\u80fd\u6e90\u6240<strong><b>\u4e1b\u5fd7\u5947\u7814\u7a76\u5458<\/b><\/strong>\u4e3a\u901a\u8baf\u4f5c\u8005\u3002\u672c\u7814\u7a76\u5f97\u5230\u56fd\u5bb6\u81ea\u7136\u79d1\u5b66\u57fa\u91d1\u548c\u56fd\u5bb6\u91cd\u70b9\u7814\u53d1\u8ba1\u5212\u9879\u76ee\u7684\u5927\u529b\u652f\u6301\u3002<\/p>\n<p>\u672c\u6587\u7248\u6743\u5c5e\u4e8e\u00a0Chem-Station\u5316\u5b66\u7a7a\u95f4\uff0c \u6b22\u8fce\u70b9\u51fb\u6309\u94ae\u5206\u4eab\uff0c\u672a\u7ecf\u8bb8\u53ef\uff0c\u8c22\u7edd\u8f6c\u8f7d<\/p>\n","protected":false},"excerpt":{"rendered":"\u4f5c\u8005\uff1a\u77f3\u6cb9\u919a \u5bfc\u8bed\uff1a \u8fd1\u65e5\uff0c\u4e2d\u56fd\u79d1\u5b66\u9662\u9752\u5c9b\u751f\u7269\u80fd\u6e90\u4e0e\u8fc7\u7a0b\u7814\u7a76\u6240\u4e1b\u5fd7\u5947\u7814\u7a76\u5458\u56e2\u961f\u901a\u8fc7\u7406\u6027\u8bbe\u8ba1P450\u9176\u7684\u6c34\u5206\u5b50\u96a7\u9053\uff08water tunnel\uff09\u6765\u4fc3\u8fdb\u8fc7\u6c27\u5316\u6c22\uff08H2O2\uff09\u8fdb\u5165\u5176\u5185\u90e8\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\uff0c\u6210\u529f\u5c06\u4e24\u79cdNADH\u4f9d\u8d56\u578bP450\u5355\u52a0\u6c27\u9176\uff08CYP199A4\u548cCYP153AM.aq\uff09\u6539\u9020\u4e3a\u5177\u6709\u9ad8\u50ac\u5316\u6d3b\u6027\u7684P450\u8fc7\u52a0\u6c27\u9176\uff0c\u5e76\u4e14\u6210\u529f\u5e94\u7528\u5230\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u534f\u540cP450BM3\u8fc7\u52a0\u6c27\u9176\u4f53\u7cfb\u3002\u96a7\u9053\u5de5\u7a0b\u6709\u671b\u6210\u4e3a\u4e00\u4e2a\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u8fc7\u52a0\u6c27\u9176\u7684\u666e\u9002\u6027\u65b0\u7b56\u7565\uff0c\u8be5\u6210\u679c\u4e8e\u8fd1\u671f\u53d1\u8868\u4e8eJ. Am. Chem. Soc.\u4e0a\u3002 \u201cEnabling Peroxygenase Activity in Cytochrome P450 Monooxygenases by Engineering Hydrogen Peroxide Tunnel. Panxia Zhao,\u2020\u00a0Fanhui Kong,\u2020\u00a0Yiping Jiang,\u2020\u00a0Xiangquan Qin, Xiaoxia Tian, and Zhiqi Cong*. J. Am. Chem. Soc.2023,ASAP.DOI 10.1021\/jacs.3c00195.\u201d \u6b63\u6587\uff1a \u7ec6\u80de\u8272\u7d20P450\u5355\u52a0\u6c27\u9176\u662f\u81ea\u7136\u754c\u4e2d\u6700\u91cd\u8981\u7684\u591a\u529f\u80fd\u751f\u7269\u6c27\u5316\u50ac\u5316\u5242\u4e4b\u4e00\uff0c\u5e7f\u6cdb\u53c2\u4e0e\u836f\u7269\u4ee3\u8c22\u3001\u5929\u7136\u4ea7\u7269\u5408\u6210\u3001\u5f02\u6e90\u7269\u8d28\u964d\u89e3\u548c\u753e\u4f53\u6fc0\u7d20\u7684\u751f\u7269\u5408\u6210\uff0c\u5e76\u4e14\u5176\u5bf9\u6781\u5177\u6311\u6218\u6027\u7684\u60f0\u6027\u78b3-\u6c22\u952e\u9009\u62e9\u6c27\u5316\u53cd\u5e94\u5177\u6709\u4f18\u5f02\u7684\u50ac\u5316\u80fd\u529b\u3002\u5927\u591a\u6570P450\u5355\u52a0\u6c27\u9176\u8981\u901a\u8fc7\u590d\u6742\u7684\u6c27\u5316\u8fd8\u539f\u4f53\u7cfb\u5b9e\u73b0\u5176\u50ac\u5316\u529f\u80fd\uff0c\u5bfc\u81f4\u5176\u5b9e\u9645\u5e94\u7528\u53d7\u5230\u4e00\u5b9a\u7684\u9650\u5236\u3002\u5229\u7528H2O2\u4ee3\u66ffO2\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u8fc7\u52a0\u6c27\u9176\uff0c\u88ab\u8ba4\u4e3a\u662f\u5f00\u53d1\u7b80\u5355\u5b9e\u7528\u578bP450\u751f\u7269\u50ac\u5316\u5242\u7684\u6709\u6548\u9014\u5f84\uff0c\u4f46\u662f\u7edd\u5927\u591a\u6570P450\u5355\u52a0\u6c27\u9176\u5229\u7528H2O2\u8fdb\u884c\u50ac\u5316\u53cd\u5e94\u4ec5\u6709\u6781\u5f31\u3001\u751a\u81f3\u6ca1\u6709\u50ac\u5316\u6d3b\u6027\u3002 \u8fd1\u5e74\u6765\uff0c\u9752\u5c9b\u80fd\u6e90\u6240\u4e1b\u5fd7\u5947\u7814\u7a76\u5458\u56e2\u961f\u81f4\u529b\u4e8eP450\u8fc7\u52a0\u6c27\u9176\u7684\u5206\u5b50\u8bbe\u8ba1\u548c\u521b\u5236\uff0c\u5728\u56fd\u9645\u4e0a\u9996\u6b21\u63d0\u51fa\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u534f\u540cP450\u9176\u50ac\u5316\u6982\u5ff5\uff0c\u6210\u529f\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u80fd\u9ad8\u6548\u5229\u7528H2O2\u7684P450\u8fc7\u52a0\u6c27\u9176\uff08Angew. Chem. Int. Ed. 2018,\u00a07628 Very Important Paper\uff09\uff0c\u5e76\u901a\u8fc7\u5408\u4f5c\u9610\u660e\u4e86\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u534f\u540cP450\u8fc7\u52a0\u6c27\u9176\u7684\u7ed3\u6784\u57fa\u7840\u548c\u50ac\u5316\u673a\u7406\uff08ACS Catal.\u00a02021, 8774\uff09\u3002\u7814\u7a76\u4eba\u5458\u8fdb\u800c\u7ed3\u5408\u86cb\u767d\u8d28\u5de5\u7a0b\u6280\u672f\uff0c\u5148\u540e\u5f00\u53d1\u51fa\u4e86H2O2\u9a71\u52a8\u7684P450\u4e19\u70f7\u7f9f\u5316\u9176\uff08ACS Catal. 2019, 7350\uff09\u3001\u9ad8\u5ea6R-\u5bf9\u6620\u9009\u62e9\u6027\u7684P450\u82ef\u4e59\u70ef\u73af\u6c27\u5316\u9176\uff08Chem. Sci. 2021, 6307\uff09\u3001\u9ad8\u6548P450\u8fc7\u6c27\u5316\u7269\u9176\uff08ACS Catal.\u00a02021, 8449\uff09\u3001\u78b3-\u6c22\u952e\u7684\u533a\u57df\u548c\u7acb\u4f53\u591a\u6837\u6027\u7f9f\u5316P450\u5de5\u7a0b\u9176\uff08Angew. Chem. Int. Ed. 2023, e202215088. Hot\u00a0Paper\uff09\u3001\u4e0d\u9971\u548c\u70c3\u76f4\u63a5\u785d\u5316\u7684P450\u785d\u5316\u9176\uff08Angew. Chem. Int. Ed. 2023, e202217678.\uff09\u3002\u4f46\u662f\u7814\u7a76\u4eba\u5458\u53d1\u73b0\u9700\u8981\u9ad8\u51fa\u5e95\u7269\u6d53\u5ea6\u6570\u500d\u7684H2O2\u624d\u80fd\u5b9e\u73b0\u9ad8\u50ac\u5316\u6d3b\u6027\uff0c\u8fd9\u5bf9\u66f4\u5177\u5b9e\u7528\u6027P450\u9176\u50ac\u5316\u5242\u7684\u5f00\u53d1\u63d0\u51fa\u4e86\u65b0\u7684\u6311\u6218\u3002 \u53e6\u5916\uff0c\u7814\u7a76\u4eba\u5458\u5229\u7528\u5b9a\u70b9\u7a81\u53d8\u3001\u5b9a\u5411\u8fdb\u5316\u548c\u53cc\u529f\u80fd\u5c0f\u5206\u5b50\u7b56\u7565\uff08\u56fe1\uff09\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u8fc7\u52a0\u6c27\u9176\uff0c\u5c06\u8fc7\u52a0\u6c27\u9176\u4e0e\u5929\u7136\u8fc7\u6c27\u5316\u7269\u9176\u7684\u7ed3\u6784\u6bd4\u5bf9\u5206\u6790\u53d1\u73b0\uff1a\u5929\u7136\u8fc7\u6c27\u5316\u7269\u9176\u7684\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\u66f4\u591a\u7684\u66b4\u9732\u4e8e\u86cb\u767d\u8868\u9762\uff0c\u6216\u8005\u6709\u8f83\u4e3a\u5f00\u653e\u7684\u5e95\u7269\u901a\u9053\u4e0e\u86cb\u767d\u8868\u9762\u76f8\u901a\uff0c\u6240\u4ee5H2O2\u5206\u5b50\u80fd\u591f\u66f4\u5bb9\u6613\u5230\u8fbe\u6d3b\u6027\u4e2d\u5fc3\u3002\u76f8\u53cd\uff0c\u70df\u9170\u80fa\u8f85\u56e0\u5b50NAD(P)H\u4f9d\u8d56\u578bP450\u9176\u7684\u6d3b\u6027\u4e2d\u5fc3\u5927\u90fd\u6df1\u57cb\u5728\u86cb\u767d\u5185\u90e8\uff0cH2O2\u5206\u5b50\u53ef\u80fd\u4e0d\u5bb9\u6613\u901a\u8fc7\u758f\u6c34\u6027\u5e95\u7269\u901a\u9053\u8fdb\u5165\u6d3b\u6027\u4e2d\u5fc3\uff0c\u5bfc\u81f4\u5de5\u7a0b\u5316P450\u8fc7\u52a0\u6c27\u9176\u5728\u8fdb\u884c\u50ac\u5316\u53cd\u5e94\u65f6\u901a\u5e38\u9700\u8981\u9ad8\u6d53\u5ea6\u7684H2O2\u3002 Figure 1. Schematic diagram of NAD(P)H-dependent P450 monooxygenases and H2O2-driven P450 peroxygenases and strategies for modifying P450 monooxygenase into its peroxygenase mode. \u9274\u4e8eP450\u9176\u7ed3\u6784\u4e2d\u5b58\u5728\u8fde\u63a5\u86cb\u767d\u8868\u9762\u4e0e\u6d3b\u6027\u4e2d\u5fc3\u7684H2O\u5206\u5b50\u96a7\u9053\uff08water tunnel\uff09\uff0c\u7814\u7a76\u4eba\u5458\u63d0\u51fa\u901a\u8fc7P450\u9176\u7684\u6c34\u96a7\u9053\u8bbe\u8ba1\u6765\u4fc3\u8fdbH2O2\u8fdb\u5165\u5176\u5185\u90e8\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\uff0c\u589e\u5f3aP450\u8fc7\u52a0\u6c27\u9176\u7684\u50ac\u5316\u6d3b\u6027\u3002\u7814\u7a76\u4eba\u5458\u5229\u7528Caver web 1.0\u9884\u6d4b\u51faP450\u9176\u8840\u7ea2\u7d20\u6d3b\u6027\u4e2d\u5fc3\u4e0e\u6eb6\u5242\u73af\u5883\u4e4b\u95f4\u7684\u6c34\u96a7\u9053\uff0c\u901a\u8fc7\u8ba1\u7b97\u5206\u6790\u9274\u5b9a\u51fa\u53ef\u80fd\u5f71\u54cdH2O2\u6d41\u5165\u6d3b\u6027\u4e2d\u5fc3\u7684\u96a7\u9053\u5165\u53e3\u6216\u74f6\u9888\u5904\u7684\u5173\u952e\u6c28\u57fa\u9178\uff0c\u7ed3\u5408\u5b9a\u70b9\u7a81\u53d8\u548c\u591a\u8f6e\u53cd\u5e94\u7b5b\u9009\uff0c\u6210\u529f\u5c06P450\u5355\u52a0\u6c27\u9176\u6539\u9020\u4e3a\u5de5\u7a0b\u5316\u7684P450\u8fc7\u52a0\u6c27\u9176\u3002 \u9996\u5148\uff0c\u7814\u7a76\u4eba\u5458\u5229\u7528\u96a7\u9053\u5de5\u7a0b\u5bf9\u6765\u81eaRhodopseudomonas palustris HaA2\u00a0NADH\u4f9d\u8d56\u7684CYP199A4\u8fdb\u884c\u7814\u7a76\u3002\u7ed3\u679c\u8868\u660eH2O\u5206\u5b50\u96a7\u9053\u74f6\u9888\u6216\u5165\u53e3\u7684\u5173\u952e\u6c28\u57fa\u9178P90\u3001F298\u548cF182\u7684\u7a81\u53d8\u6709\u8f83\u4f18\u50ac\u5316\u6027\u80fd\uff08\u56fe2A\uff09\uff0cF182A\u50ac\u53164-\u7532\u6c27\u57fa\u82ef\u7532\u9178O-\u8131\u7532\u57fa\u5316\u6d3b\u6027\u76f8\u6bd4\u91ce\u751f\u578b\u63d0\u5347\u4e86163\u500d\uff08\u56fe2B\uff09\uff0c\u5176\u50ac\u5316\u8f6c\u5316\u6570\uff08TON\uff09\u4e3a980\u00b5M(\u00b5M P450)-1\u3002\u968f\u540e\uff0c\u7814\u7a76\u4eba\u5458\u5c06\u6709\u76ca\u7a81\u53d8\u4f53\u8fdb\u884c\u7ec4\u5408\u7a81\u53d8\uff0c\u53d1\u73b0\u53cc\u7a81\u53d8F182A\/P90A\u7684TON\u8fbe\u52301100\u00b5M(\u00b5M P450)-1(\u589e\u52a0183\u500d)\uff08\u56fe2B\uff09\u3002 \u5728\u4e0a\u8ff0\u57fa\u7840\u4e0a\uff0c\u7814\u7a76\u4eba\u5458\u4e3a\u4e86\u8fdb\u4e00\u6b65\u9a8c\u8bc1\u8be5\u7b56\u7565\u7684\u666e\u9002\u6027\uff0c\u5c06\u4e0a\u8ff0\u7b56\u7565\u5e94\u7528\u4e8e\u6765\u81eaMarinobacter aquaeolei\u00a0\u7684NADH\u4f9d\u8d56\u7684CYP153AM.aq\u3002\u7ed3\u679c\u8868\u660e\uff1a\u4e0e\u91ce\u751f\u578b\u76f8\u6bd4\uff0c\u76f8\u540c\u96a7\u9053\u5185\u7684\u5173\u952e\u6c28\u57fa\u9178D226\u3001D310\u548cV456\u50ac\u5316\u6708\u6842\u9178\u7684\u7f9f\u5316\u6d3b\u6027\u63d0\u5347\u81f313\u500d\u4ee5\u4e0a\uff08\u56fe2D-2E\uff09\uff0c\u5c06\u4e0a\u8ff0\u5173\u952e\u6c28\u57fa\u9178\u8fdb\u884c\u7ec4\u5408\u7a81\u53d8\uff0c\u53d1\u73b0\u53cc\u7a81\u53d8\u4f53D266A\/V456A\u548cD266T\/V456G\u7684TON\u5206\u522b\u63d0\u9ad8\u81f3161\u00b5M\u548c167\u00b5M(\u00b5M P450)-1\uff0c\u6bd4WT\u63d0\u9ad815\u500d\uff08\u56fe2E\uff09\u3002 Figure 2. Engineering the peroxygenase activity of CYP199A4 and CYP153AM.aq. 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