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It could be enzymatically transformed into 10\u2010deacetyltaxol (DT), and the latter could be further converted into Taxol. The enzyme 10\u2010deacetylbaccatin III\u201010\u2010\u03b2\u2010O\u2010acetyltransferase (DBAT) can acetylate the non\u2010natural substrate DT into Taxol, but the conversion efficiency was extremely low. Herein, we globally redesigned DBAT\n                    <jats:sub>cus<\/jats:sub>\n                    from\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Taxus cuspidata<\/jats:italic>\n                    <\/jats:styled-content>\n                    to improve its efficiency in DT acetylation through combinatorial protein engineering strategies including virtual saturation mutagenesis, in silico screening, DNA shuffling, and iterative combinatorial mutagenesis. Several more active DBAT mutants against DT were obtained, among which the ICM9\u20106 exhibited 16.4 times higher activity than DBAT\n                    <jats:sub>cus<\/jats:sub>\n                    . The transient expression system of\n                    <jats:italic>Nicotiana benthamiana<\/jats:italic>\n                    was then established, and the ICM9\u20106 was functionally expressed in the system, with yield of 8.2\u2009\u03bcg\u2009g\n                    <jats:sup>\u22121<\/jats:sup>\n                    FW (129.3\u2009\u03bcg\u2009g\n                    <jats:sup>\u22121<\/jats:sup>\n                    DW) Taxol when the system was fed with DT. Specifically, the fungal glycoside hydrolase LXYL\u2010P1\u20102 that was responsible for converting XDT into DT was also functionally expressed in the system, and upon feeding XDT, the co\u2010expression of LXYL\u2010P1\u20102 and ICM9\u20106 yielded 3.6\u2009\u03bcg\u2009g\n                    <jats:sup>\u22121<\/jats:sup>\n                    FW (55.4\u2009\u03bcg\u2009g\n                    <jats:sup>\u22121<\/jats:sup>\n                    DW) Taxol. These results represent the highest reported Taxol productivity in the tobacco system to date and lay a foundation for the construction of the stable transgenic cell lines of tobacco and more efficiently converting DT or XDT into Taxol for the large\u2010scale pharmaceutical manufacturing.\n                  <\/jats:p>","DOI":"10.1111\/pbi.70566","type":"journal-article","created":{"date-parts":[[2026,1,31]],"date-time":"2026-01-31T07:22:28Z","timestamp":1769844148000},"page":"3308-3323","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Engineering of 10\u2010Deacetylbaccatin III\u201010\u2010\u03b2\u2010O\u2010Acetyltransferase From\n                    <i>Taxus<\/i>\n                    Species for Efficient Acetylating Non\u2010Natural Substrates Into Taxol in\n                    <i>Nicotiana benthamiana<\/i>"],"prefix":"10.1111","volume":"24","author":[{"given":"Tian\u2010Jiao","family":"Chen","sequence":"first","affiliation":[{"name":"State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs Institute of Materia Medica, Chinese Academy of Medical Sciences &amp; Peking Union Medical College  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing\u2010Jing","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs Institute of Materia Medica, Chinese Academy of Medical Sciences &amp; Peking Union Medical College  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao\u2010Yan","family":"Sun","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs Institute of Materia Medica, Chinese Academy of Medical Sciences &amp; Peking Union Medical College  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting","family":"Gong","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs Institute of Materia Medica, Chinese Academy of Medical Sciences &amp; Peking Union Medical College  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin\u2010Ling","family":"Yang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs Institute of Materia Medica, Chinese Academy of Medical Sciences &amp; Peking Union Medical College  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Zhu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs Institute of Materia Medica, Chinese Academy of Medical Sciences &amp; Peking Union Medical College  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,1,30]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.00218"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.2174\/1381612829666221102155212"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1191652"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1515826113"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jcim.3c01153"},{"key":"e_1_2_9_7_1","unstructured":"Chattopadhyay S. K. R. P.Sharma S.Kumar andK. P.Madhusudanan.2002.\u201cA Process for the Production of Taxol. European Patent 0905130 B1.\u201d"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.M113.030619"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molp.2021.04.015"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.20694"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja00225a063"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep13768"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/pca.2800040307"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1039\/c2np20021j"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1470-8744.1997.tb01323.x"},{"key":"e_1_2_9_16_1","unstructured":"Hanson R. L. R. N.Patel andL. J.Szarka.1997.\u201cEnzymatic Hydrolysis Method for the Conversion of C\u20107 Sugar to C\u20107 Hydroxyl Taxanes. 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