State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, P.R. China.
Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing Tech University, Nanjing 211800, P.R. China.
J Ind Microbiol Biotechnol. 2023 Feb 17;50(1). doi: 10.1093/jimb/kuad009.
β-Carotene is a kind of high-value tetraterpene compound, which shows various applications in medical, agricultural, and industrial areas owing to its antioxidant, antitumor, and anti-inflammatory activities. In this study, Yarrowia lipolytica was successfully metabolically modified through the construction and optimization of β-carotene biosynthetic pathway for β-carotene production. The β-carotene titer in the engineered strain Yli-C with the introduction of the carotenogenesis genes crtI, crtE, and crtYB can reach 34.5 mg/L. With the overexpression of key gene in the mevalonate pathway and the enhanced expression of the fatty acid synthesis pathway, the β-carotene titer of the engineered strain Yli-CAH reached 87 mg/L, which was 152% higher than that of the strain Yli-C. Through the further expression of the rate-limiting enzyme tHMGR and the copy number of β-carotene synthesis related genes, the β-carotene production of Yli-C2AH2 strain reached 117.5 mg/L. The final strain Yli-C2AH2 produced 2.7 g/L β-carotene titer by fed-batch fermentation in a 5.0-L fermenter. This research will greatly speed up the process of developing microbial cell factories for the commercial production of β-carotene.
ONE-SENTENCE SUMMARY: In this study, the β-carotene synthesis pathway in engineered Yarrowia lipolytica was enhanced, and the fermentation conditions were optimized for high β-carotene production.
β-胡萝卜素是一种高附加值的四萜化合物,由于其抗氧化、抗肿瘤和抗炎活性,在医学、农业和工业领域显示出各种应用。在这项研究中,通过构建和优化β-胡萝卜素生物合成途径,成功地对产脂酵母进行了代谢修饰,以生产β-胡萝卜素。引入类胡萝卜素生物合成基因 crtI、crtE 和 crtYB 的工程菌株 Yli-C 的β-胡萝卜素产量可达 34.5mg/L。通过过表达甲羟戊酸途径中的关键基因和增强脂肪酸合成途径的表达,工程菌株 Yli-CAH 的β-胡萝卜素产量达到 87mg/L,比菌株 Yli-C 提高了 152%。通过进一步表达限速酶 tHMGR 和β-胡萝卜素合成相关基因的拷贝数,菌株 Yli-C2AH2 的β-胡萝卜素产量达到 117.5mg/L。最终菌株 Yli-C2AH2 在 5.0L 发酵罐中通过分批补料发酵生产 2.7g/L 的β-胡萝卜素产量。这项研究将大大加快开发微生物细胞工厂用于商业生产β-胡萝卜素的进程。
本研究对工程化产脂酵母中的β-胡萝卜素合成途径进行了增强,并对发酵条件进行了优化,以实现高β-胡萝卜素产量。