Carrageenan oligosaccharides have shown promising bioavailability and possess a variety of physiological activities, making them highly suitable for use in the food, pharmaceutical, and agricultural industries. The preferred method for producing carrageenan oligosaccharides is using various carrageenolytic enzymes, as it offers mild reaction conditions, high efficiency, and product specificity. However, there is still a lack of specific applications for using these enzymes to prepare odd-numbered carrageenan-oligosaccharides (OCOSs). Our previous research identified a more convenient route for simultaneously preparing OCOSs and 3,6-anhydro-D-galactose (D-AHG) using only two types of carrageenolytic enzymes: κ-carrageenase and exo-α-3,6-anhydro-D-galactosidase (D-ADAGase). In this study, we utilized a CipA-based self-assembly system to cascade κ-carrageenase CaKC16A and D-ADAGase ZuGH129A for one-step preparation of β/κ-carrapentaose, G-(DA-G4S)₂, and D-AHG from degrading β/κ-carrageenan. This self-assembled enzyme, namely CipA-CaKC16A-ZuGH129A, can be easily obtained through a simple centrifugation process. The final optimized enzymatic process produced 0.74 g/L G-(DA-G4S)₂ and 0.13 g/L D-AHG. This cascade system of different types of carrageenolytic enzymes has the potential to achieve the preparation of various types of carrageenan oligosaccharides.

中文翻译：

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通过级联 κ-角叉菜酶和 Exo-α-3,6-Anhydro-D-半乳糖苷酶同时一步法制备 β/κ-Carrapentaose 和 3,6-Anhydro-D-半乳糖苷酶


卡拉胶低聚糖已显示出良好的生物利用度，并具有多种生理活性，使其非常适合用于食品、制药和农业工业。生产角叉菜胶低聚糖的首选方法是使用各种角叉菜胶溶解酶，因为它提供温和的反应条件、高效率和产品特异性。然而，使用这些酶制备奇数角叉菜胶寡糖 （OCOS） 仍然缺乏特异性应用。我们之前的研究确定了一种更方便的途径，可以同时使用两种类型的角叉菜酶同时制备 OCOS 和 3,6-脱水-D-半乳糖 （D-AHG）：κ-角叉菜酶和外α-3,6-脱水-D-半乳糖苷酶 （D-ADAGase）。在这项研究中，我们利用基于 CipA 的自组装系统级联 κ-角叉菜酶 CaKC16A 和 D-ADAGase ZuGH129A，用于从降解的 β/κ-角叉菜胶中一步制备 β/κ-角叉菜胶、G-（DA-G4S）₂ 和 D-AHG。这种自组装酶，即 CipA-CaKC16A-ZuGH129A，可以通过简单的离心过程轻松获得。最终优化的酶法工艺产生了 0.74 g/L G-（DA-G4S）₂ 和 0.13 g/L D-AHG。这种由不同类型的角叉菜胶分解酶组成的级联系统有可能实现制备各种类型的角叉菜胶低聚糖。