10-Deacetylbaccatin III (10-DAB) C10 acetylation is an indispensable procedure for Taxol semi-synthesis, which often requires harsh conditions. 10-Deacetylbaccatin III-10-β-O-acetyltransferase (DBAT) catalyzes the acetylation but acetyl-CoA supply remains a key limiting factor. Here we refactored the innate biosynthetic pathway of acetyl-CoA in Escherichia coli and obtained a chassis with acetyl-CoA productivity over three times higher than that of the host cell. Then, we constructed a microbial cell factory by introducing DBAT gene into this chassis for efficiently converting 10-DAB into baccatin III. We found that baccatin III could be efficiently deacetylated into 10-DAB by DBAT with CoASH and K⁺ under alkaline condition. Thus, we fed acetic acid to the engineered strain both for serving as a substrate of acetyl-CoA biosynthesis and for alleviating the deacetylation of baccatin III. The fermentation conditions were optimized and the baccatin III titers reached 2, 3 and 4.6 g/L, respectively, in a 3-L bioreactor culture when 2, 3 and 6 g/L of 10-DAB were supplied. Our study provides an environment-friendly approach for the large scale 10-DAB acetylation without addition of acetyl-CoA in the industrial Taxol semi-synthesis. The finding of DBAT deacetylase activity may broaden its application in the structural modification of pharmaceutically important lead compounds.

10-脱乙酰浆果赤霉素 III (10-DAB) C10 乙酰化是紫杉醇半合成必不可少的过程，通常需要苛刻的条件。10-脱乙酰浆果赤霉素III-10- β - O-乙酰转移酶 (DBAT) 催化乙酰化，但乙酰辅酶 A 供应仍然是一个关键的限制因素。在这里，我们重构了大肠杆菌中乙酰辅酶A的先天生物合成途径，并获得了乙酰辅酶A生产力比宿主细胞高出三倍以上的底盘。然后，我们通过将 DBAT 基因引入该底盘构建了微生物细胞工厂，以将 10-DAB 高效转化为浆果赤霉素 III。我们发现 baccatin III 可以通过 DBAT 与 CoASH 和 K ⁺有效脱乙酰成 10-DAB在碱性条件下。因此，我们将乙酸喂给工程菌株，作为乙酰辅酶 A 生物合成的底物和减轻浆果赤霉素 III 的脱乙酰作用。当提供 2、3 和 6 g/L 的 10-DAB 时，优化发酵条件，在 3-L 生物反应器培养物中，浆果赤霉素 III 滴度分别达到 2、3 和 4.6 g/L。我们的研究为在工业紫杉醇半合成中不添加乙酰辅酶 A 的大规模 10-DAB 乙酰化提供了一种环境友好的方法。DBAT 脱乙酰酶活性的发现可能会拓宽其在药学上重要的先导化合物的结构修饰中的应用。