The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, College of Life Sciences, Nankai University, Tianjin, 300071, China.
College Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, China.
Microb Cell Fact. 2022 May 10;21(1):81. doi: 10.1186/s12934-022-01805-5.
Aromatic compounds, such as p-coumaric acid (p-CA) and caffeic acid, are secondary metabolites of various plants, and are widely used in diet and industry for their biological activities. In addition to expensive and unsustainable methods of plant extraction and chemical synthesis, the strategy for heterologous synthesis of aromatic compounds in microorganisms has received much attention. As the most abundant renewable resource in the world, lignocellulose is an economical and environmentally friendly alternative to edible, high-cost carbon sources such as glucose.
In the present study, carboxymethyl-cellulose (CMC) was utilized as the sole carbon source, and a metabolically engineered Saccharomyces cerevisiae strain SK10-3 was co-cultured with other recombinant S. cerevisiae strains to achieve the bioconversion of value-added products from CMC. By optimizing the inoculation ratio, interval time, and carbon source content, the final titer of p-CA in 30 g/L CMC medium was increased to 71.71 mg/L, which was 155.9-fold higher than that achieved in mono-culture. The de novo biosynthesis of caffeic acid in the CMC medium was also achieved through a three-strain co-cultivation. Caffeic acid production was up to 16.91 mg/L after optimizing the inoculation ratio of these strains.
De novo biosynthesis of p-CA and caffeic acid from lignocellulose through a co-cultivation strategy was achieved for the first time. This study provides favorable support for the biosynthesis of more high value-added products from economical substrates. In addition, the multi-strain co-culture strategy can effectively improve the final titer of the target products, which has high application potential in the field of industrial production.
芳香族化合物,如对香豆酸(p-CA)和咖啡酸,是各种植物的次生代谢物,由于其生物活性,被广泛应用于饮食和工业。除了昂贵且不可持续的植物提取和化学合成方法外,微生物中芳香族化合物的异源合成策略受到了广泛关注。木质纤维素作为世界上最丰富的可再生资源,是一种经济且环保的替代物,可以替代葡萄糖等昂贵的、不可持续的食用型高成本碳源。
在本研究中,羧甲基纤维素(CMC)被用作唯一的碳源,同时将代谢工程化的酿酒酵母 SK10-3 菌株与其他重组酿酒酵母菌株共培养,以实现 CMC 中附加值产品的生物转化。通过优化接种比例、间隔时间和碳源含量,最终在 30g/L CMC 培养基中 p-CA 的产量提高到 71.71mg/L,比单培养提高了 155.9 倍。通过三菌株共培养还在 CMC 培养基中实现了咖啡酸的从头生物合成。优化这些菌株的接种比例后,咖啡酸的产量达到 16.91mg/L。
首次通过共培养策略实现了木质纤维素中 p-CA 和咖啡酸的从头生物合成。该研究为从经济基质中生物合成更多高附加值产品提供了有利支持。此外,多菌株共培养策略可以有效提高目标产物的最终产量,在工业生产领域具有很高的应用潜力。