Department of Chemical Engineering, Osaka Metropolitan University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka, 599-8531, Japan.
World J Microbiol Biotechnol. 2024 Jun 3;40(7):230. doi: 10.1007/s11274-024-04037-4.
β-Carotene is an attractive compound and that its biotechnological production can be achieved by using engineered Saccharomyces cerevisiae. In a previous study, we developed a technique for the efficient establishment of diverse mutants through the introduction of point and structural mutations into the yeast genome. In this study, we aimed to improve β-carotene production by applying this mutagenesis technique to S. cerevisiae strain that had been genetically engineered for β-carotene production. Point and structural mutations were introduced into β-carotene-producing engineered yeast. The resulting mutants showed higher β-carotene production capacity than the parental strain. The top-performing mutant, HP100_74, produced 37.6 mg/L of β-carotene, a value 1.9 times higher than that of the parental strain (20.1 mg/L). Gene expression analysis confirmed an increased expression of multiple genes in the glycolysis, mevalonate, and β-carotene synthesis pathways. In contrast, expression of ERG9, which functions in the ergosterol pathway competing with β-carotene production, was decreased in the mutant strain. The introduction of point and structural mutations represents a simple yet effective method for achieving mutagenesis in yeasts. This technique is expected to be widely applied in the future to produce chemicals via metabolic engineering of S. cerevisiae.
β-胡萝卜素是一种有吸引力的化合物，其生物技术生产可以通过使用工程化的酿酒酵母来实现。在之前的研究中，我们开发了一种通过向酵母基因组中引入点突变和结构突变来有效建立多种突变体的技术。在这项研究中，我们旨在通过将这种诱变技术应用于已经过基因工程改造以生产β-胡萝卜素的酿酒酵母菌株，来提高β-胡萝卜素的产量。我们向生产β-胡萝卜素的工程化酵母中引入了点突变和结构突变。与亲本菌株相比，产生的突变体显示出更高的β-胡萝卜素生产能力。表现最好的突变体 HP100_74 产生了 37.6mg/L 的β-胡萝卜素，比亲本菌株（20.1mg/L）高 1.9 倍。基因表达分析证实，在糖酵解、甲羟戊酸和β-胡萝卜素合成途径中，多个基因的表达增加。相比之下，在突变菌株中，与β-胡萝卜素生产竞争的功能在麦角固醇途径中的 ERG9 的表达降低。点突变和结构突变的引入代表了一种简单而有效的方法，可以在酵母中实现诱变。该技术有望在未来被广泛应用于通过酿酒酵母的代谢工程来生产化学品。
