{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T14:35:58Z","timestamp":1786977358525,"version":"build-2736575974"},"reference-count":39,"publisher":"Wiley","issue":"14","license":[{"start":{"date-parts":[[2024,6,27]],"date-time":"2024-06-27T00:00:00Z","timestamp":1719446400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2024,7,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    \u03c9\u2010Transaminases (\u03c9\u2010TAs) are attractive biocatalysts asymmetrically catalyzing ketones to chiral amines. However, poor non\u2010native catalytic activity and substrate promiscuity severely hamper its wide application in industrial production. Protein engineering efforts have generally focused on reshaping the substrate\u2010binding pockets of \u03c9\u2010TAs. However, hotspots around the substrate tunnel as well as distant sites outside the pockets may also affect its activity. In this study, the \u03c9\u2010TA from\n                    <jats:italic>Bacillus megaterium<\/jats:italic>\n                    (\n                    <jats:italic>Bme<\/jats:italic>\n                    TA) was selected for engineering. The tunnel mutation Y164F synergy with distant mutation A245T which was acquired through a multiple sequence alignment showed improved soluble expression, a 3.7\u2010fold higher specific activity and a 19.9\u2010fold longer half\u2010life at 45\u2009\u00b0C. Molecule Dynamics simulation explains the mechanism of improved catalytic activity, enhanced thermostability and improved soluble expression of\n                    <jats:italic>Bme<\/jats:italic>\n                    TA\n                    <jats:sup>Y164F\/A245T<\/jats:sup>\n                    (2\u2005M). Finally, the resting cells of 2\u2005M were used for biocatalytic processes. 450\u2005mM of\n                    <jats:italic>S<\/jats:italic>\n                    \u2010methoxyisopropylamine (\n                    <jats:italic>S<\/jats:italic>\n                    \u2010MOIPA) was obtained with an\n                    <jats:italic>ee<\/jats:italic>\n                    value of 97.3\u2009% and a conversion rate of 90\u2009%, laying the foundation for its industrial production. Mutant 2\u2005M was also found to be more advantageous in catalyzing the transamination of various ketones. These results demonstrated that sites that are far away from the active center also play an important role in the redesign of \u03c9\u2010TAs.\n                  <\/jats:p>","DOI":"10.1002\/cbic.202400285","type":"journal-article","created":{"date-parts":[[2024,5,16]],"date-time":"2024-05-16T10:18:49Z","timestamp":1715854729000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Sequence\u2010Guided Redesign of an Omega\u2010Transaminase from\n                    <i>Bacillus megaterium<\/i>\n                    for the Asymmetric Synthesis of Chiral Amines"],"prefix":"10.1002","volume":"25","author":[{"given":"Zhexian","family":"Xu","sequence":"first","affiliation":[{"name":"Institute of Bioengineering College of Chemical and Biological Engineering Zhejiang University  Hangzhou 310027 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7500-882X","authenticated-orcid":false,"given":"Jiaqi","family":"Xu","sequence":"additional","affiliation":[{"name":"Institute of Bioengineering College of Chemical and Biological Engineering Zhejiang University  Hangzhou 310027 China"},{"name":"Hangzhou Global Scientific and Technological Innovation Center Zhejiang University  Hangzhou 310027 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute of Bioengineering College of Chemical and Biological Engineering Zhejiang University  Hangzhou 310027 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyuan","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Bioengineering College of Chemical and Biological Engineering Zhejiang University  Hangzhou 310027 China"},{"name":"Hangzhou Global Scientific and Technological Innovation Center Zhejiang University  Hangzhou 310027 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianping","family":"Wu","sequence":"additional","affiliation":[{"name":"Institute of Bioengineering College of Chemical and Biological Engineering Zhejiang University  Hangzhou 310027 China"},{"name":"Hangzhou Global Scientific and Technological Innovation Center Zhejiang University  Hangzhou 310027 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lirong","family":"Yang","sequence":"additional","affiliation":[{"name":"Institute of Bioengineering College of Chemical and Biological Engineering Zhejiang University  Hangzhou 310027 China"},{"name":"Hangzhou Global Scientific and Technological Innovation Center Zhejiang University  Hangzhou 310027 China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,6,27]]},"reference":[{"key":"e_1_2_8_1_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201005721"},{"key":"e_1_2_8_2_1","first-page":"510","volume":"31","author":"Yang J.","year":"2014","journal-title":"Chin. 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