Zhong Jialing, Wang Yushu, Chen Zhuoyang, Yalikun Yaliqin, He Lin, Liu Tiangang, Ma Gang
Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), Shanghai Jiao Tong University, No. 800 Dongchuan Rd., Shanghai, 200240, People's Republic of China.
School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
Biotechnol Biofuels Bioprod. 2024 Jul 16;17(1):99. doi: 10.1186/s13068-024-02555-9.
Taxol serves as an efficient natural anticancer agent with extensive applications in the treatment of diverse malignancies. Although advances in synthetic biology have enabled the de novo synthesis of taxol precursors in various microbial chassis, the total biosynthesis of taxol remains challengable owing to the restricted oxidation efficiency in heterotrophic microbes. Here, we engineered Synechocystis sp. PCC 6803 with modular metabolic pathways consisting of the methylerythritol phosphate pathway enzymes and taxol biosynthetic enzymes for production of taxadiene-5α-ol (T5α-ol), the key oxygenated intermediate of taxol. The best strain DIGT-P560 produced up to 17.43 mg/L of oxygenated taxanes and 4.32 mg/L of T5α-ol. Moreover, transcriptomic analysis of DIGT-P560 revealed that establishing a oxygenated taxane flux may enhance photosynthetic electron transfer efficiency and central metabolism in the engineered strain to ameliorate the metabolic disturbances triggered by the incorporation of exogenous genes. This is the first demonstration of photosynthetic production of taxadiene-5α-ol from CO in cyanobacteria, highlighting the broad prospects of engineered cyanobacteria as bio-solar cell factories for valuable terpenoids production and expanding the ideas for further rational engineering and optimization.
紫杉醇是一种高效的天然抗癌药物，在多种恶性肿瘤的治疗中有着广泛应用。尽管合成生物学的进展已使紫杉醇前体能够在各种微生物底盘中从头合成，但由于异养微生物中氧化效率受限，紫杉醇的全生物合成仍然具有挑战性。在此，我们对集胞藻PCC 6803进行了工程改造，构建了由甲基赤藓糖醇磷酸途径酶和紫杉醇生物合成酶组成的模块化代谢途径，用于生产紫杉二烯-5α-醇（T5α-ol），这是紫杉醇的关键氧化中间体。最佳菌株DIGT-P560产生了高达17.43毫克/升的氧化紫杉烷和4.32毫克/升的T5α-醇。此外，对DIGT-P560的转录组分析表明，建立氧化紫杉烷通量可能会提高工程菌株的光合电子传递效率和中心代谢，以改善由外源基因整合引发的代谢紊乱。这是首次证明蓝藻能从二氧化碳光合生产紫杉二烯-5α-醇，突出了工程化蓝藻作为生产有价值萜类化合物的生物太阳能电池工厂的广阔前景，并拓展了进一步进行合理工程设计和优化的思路。
