Artificial Cascade Transformation Biosystem for One-Pot Biomanufacturing of Odd-Numbered Neoagarooligosaccharides and d -Tagatose through Wiser Agarose Utilization

作者：Xiaohan Xu, Jing Gao, Liwei Qing, Mingyue Zhang, Jianan Sun, Hong Jiang, Sai Wang, Hao Dong, Xiangzhao Mao · 发表于：Journal of Agricultural and Food Chemistry · 年份：2024 · DOI：10.1021/acs.jafc.4c08619 · 被引用次数：4 · 研究领域：Diet, Metabolism, and Disease、Pancreatic function and diabetes、Microbial Metabolites in Food Biotechnology

The application of agarose oligosaccharides has garnered great attention, with their biological activities varying among different structures. However, it still meets a great bottleneck for the targeted production of odd-numbered neoagarooligosaccharides (NAOSs), such as neoagarotriose (NA3), due to the lack of one-step hydrolases. In this work, the α-agarase AgaA33 and β-galactosidase BgaD were synergistically used to prepare NA3 with agarose as a substrate. Additionally, an l -arabinose isomerase CaLAI from Clostridium acetobutylicum was characterized to valorize low-value byproducts ( d -galactose) by forming d -tagatose, which exhibited good thermal stability without the need for additional metal ions. Under the optimal reaction conditions, the production of NA3 and d -galactose catalyzed by these three enzymes was 0.40 and 0.15 g/L, respectively. The artificial three-enzyme-based cascade transformation system not only achieved the highest production of NA3 until now but also allowed for the valorization of d -galactose, providing a wiser application route for agarose utilization.