Arenas Natalia, Cataldo Vicente F, Agosin Eduardo
Department of Chemical and Bioprocess Engineering, School of Engineering, Pontificia Universidad Católica de Chile, Av. Vicuña Mackenna 4860, 7820436, Santiago, Chile.
Sticta Biologicals, Santiago, Chile.
Microb Cell Fact. 2025 Aug 1;24(1):176. doi: 10.1186/s12934-025-02789-8.
Xanthophylls, a subclass of oxygenated carotenoids, are highly valued for their wide range of applications in the food and pharmaceutical industries, particularly due to their antioxidant properties and potential health benefits. Among these, neoxanthin, a less studied xanthophyll, has demonstrated significant therapeutic potential, including antioxidant and anticancer activities. Neoxanthin is also the primary precursor for the synthesis of other valuable compounds, such as fucoxanthin and β-damascenone, which are important in the cosmetic and pharmaceutical sectors.
In this study, we report the first heterologous production of neoxanthin in Saccharomyces cerevisiae through a combination of metabolic and enzyme engineering. First, a S. cerevisiae strain was engineered to produce neoxanthin by expressing genes from the β-carotene and violaxanthin biosynthesis pathways. Following this, the VDL1 gene from Phaeodactylum tricornutum, responsible for converting violaxanthin into neoxanthin, was expressed, resulting in the production of 0.18 mg/g of neoxanthin. To further enhance production, a pulse-fed galactose strategy was employed during shake-flask growth, leading to a 2.5-fold increase in neoxanthin yield. Additionally, transmembrane peptides were incorporated into the yeast cells to improve the accumulation of carotenoids, generating an increase of 3.8-fold, achieving a final production of 0.7 mg/g of neoxanthin.
This is the highest reported yield of neoxanthin produced by engineered microorganisms, and the strategies employed here have considerable potential for scaling up production of this carotenoid.
叶黄素是氧化类胡萝卜素的一个亚类，因其在食品和制药行业的广泛应用而备受重视，特别是由于其抗氧化特性和潜在的健康益处。其中，新黄质作为一种研究较少的叶黄素，已显示出显著的治疗潜力，包括抗氧化和抗癌活性。新黄质也是合成其他有价值化合物的主要前体，如岩藻黄质和β-大马酮，它们在化妆品和制药领域很重要。
在本研究中，我们报告了通过代谢和酶工程相结合的方法首次在酿酒酵母中异源生产新黄质。首先，通过表达来自β-胡萝卜素和紫黄质生物合成途径的基因，构建了一株能生产新黄质的酿酒酵母菌株。随后，表达了来自三角褐指藻的负责将紫黄质转化为新黄质的VDL1基因，从而产生了0.18毫克/克的新黄质。为了进一步提高产量，在摇瓶培养过程中采用了脉冲补加半乳糖策略，使新黄质产量提高了2.5倍。此外，将跨膜肽整合到酵母细胞中以改善类胡萝卜素的积累，产量提高了3.8倍，最终新黄质产量达到0.7毫克/克。
这是工程微生物生产新黄质的最高报道产量，此处采用的策略在扩大这种类胡萝卜素的生产方面具有相当大的潜力。
