Rehman Esha, Birla Singh Hawaibam, Nguyen Minh Phuong, Wang Chonglong, Yoon Sang-Hwal, Kwon Moonhyuk, Kang Min-Kyoung, Kim Seon-Won
Anti-Aging Bio Cell Factory Regional Leading Research Center, Gyeongsang National University, Jinju, Republic of Korea.
Division of Applied Life Science (BK21 Four), Gyeongsang National University, Jinju, Republic of Korea.
Front Bioeng Biotechnol. 2025 Aug 29;13:1670015. doi: 10.3389/fbioe.2025.1670015. eCollection 2025.
a Generally Recognized As Safe (GRAS) microorganism, is an attractive chassis for producing high-value compounds in a safe and sustainable way. However, its potential for producing the C40 carotenoid lycopene has been limited by inefficient precursor supply and enzyme incompatibility. This study demonstrates that lycopene production in can be significantly enhanced through systematic metabolic engineering by rewiring the lycopene and methylerythritol phosphate (MEP) pathways. A synthetic lycopene biosynthesis pathway expressing the gene from , which is commonly used for microbial lycopene production, failed to yield lycopene production in . However, replacing with a multifunctional geranylgeranyl diphosphate synthase (GGPPS) from successfully enabled lycopene synthesis. The optimization of the fermentation medium demonstrated that a combined carbon supply of glucose and glycerol markedly enhanced both cell growth and lycopene production in comparison with separate carbon sources. To further boost production, the methylerythritol phosphate (MEP) pathway was engineered by overexpressing the rate-limiting enzyme, 1-deoxy-D-xylulose-5-phosphate synthase (), which resulted in a five-fold increase in lycopene titer after 72 h. Screening of various GGPPS enzymes revealed that from was the most efficient, further increasing the yield. The final engineered strain achieved a lycopene titer of 55 mg/L in shake-flask cultivation, a significant improvement over the previously reported level in . These results demonstrate that targeted GGPPS selection and precursor pathway engineering are critical strategies for developing into a robust and sustainable platform for carotenoid production.
一种普遍认为安全（GRAS）的微生物，是一种以安全和可持续的方式生产高价值化合物的有吸引力的底盘。然而，其生产C40类胡萝卜素番茄红素的潜力受到前体供应效率低下和酶不兼容性的限制。本研究表明，通过对番茄红素和甲基赤藓糖醇磷酸（MEP）途径进行重新布线的系统代谢工程，可以显著提高番茄红素的产量。一个表达常用于微生物番茄红素生产的来自[具体物种]的[基因名称]的合成番茄红素生物合成途径，在[具体物种]中未能产生番茄红素。然而，用来自[具体物种]的多功能牻牛儿基牻牛儿基二磷酸合酶（GGPPS）取代[具体基因]成功实现了番茄红素的合成。发酵培养基的优化表明，与单独的碳源相比，葡萄糖和甘油的组合碳供应显著提高了细胞生长和番茄红素产量。为了进一步提高产量，通过过表达限速酶1-脱氧-D-木酮糖-5-磷酸合酶（[酶名称]）对甲基赤藓糖醇磷酸（MEP）途径进行工程改造，这导致72小时后番茄红素滴度增加了五倍。对各种GGPPS酶的筛选表明，来自[具体物种]的[酶名称]是最有效的，进一步提高了产量。最终的工程菌株在摇瓶培养中达到了55mg/L的番茄红素滴度，比之前报道的[具体物种]水平有显著提高。这些结果表明，靶向GGPPS选择和前体途径工程是将[具体物种]发展成为一个强大且可持续的类胡萝卜素生产平台的关键策略。
