Isabel Katja Sommerfeld, Esther Maria Dälken, Lothar Elling, Andrij Pich
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引用次数: 0
摘要
酶在合成透明质酸(HA)等复杂生物分子方面发挥着重要作用。将酶固定在支撑材料上对酶的高效利用和多次循环再利用至关重要。微凝胶由交联、高度膨胀的聚合物网络组成,因其孔隙率高而非常适合酶的吸收。本研究利用不同的二价离子(Ni2+、Co2+、Mn2+、Mg2+ 和 Fe2+),通过金属亲和性结合将 His6 标记的多杀性巴氏杆菌透明质酸合成酶(PmHAS)固定在氮基三乙酸功能化微凝胶上。结果表明,使用 Ni2+ 得到的微凝胶具有最高的酶吸收率和 HA 形成率。固定化 PmHAS 可实现重复酶解生产,生产出高分子量的 HA,且每一步的分散度都在降低。此外,据报道,固定化 PmHAS 的高分子量 HA 产量最高。该系统为连续 HA 的形成奠定了基础,未来的工作有可能通过蛋白质工程提高 PmHAS 的稳定性。
Nitrilotriacetic Acid Functionalized Microgels for Efficient Immobilization of Hyaluronan Synthase
Enzymes play a vital role in synthesizing complex biological molecules like hyaluronic acid (HA). Immobilizing enzymes on support materials is essential for their efficient use and reuse in multiple cycles. Microgels, composed of cross–linked, highly swollen polymer networks, are ideal for enzyme uptake owing to their high porosity. This study demonstrates the immobilization of His6-tagged hyaluronan synthase from Pasteurella multocida (PmHAS) onto nitrilotriacetic acid functionalized microgels using different bivalent ions (Ni2+, Co2+, Mn2+, Mg2+, and Fe2+) via metal affinity binding. The results indicate that using Ni2+ yields the microgels with the highest enzyme uptake and HA formation. The immobilized PmHAS enables repetitive enzymatic production, producing high molecular weight HAs with decreasing dispersities in each step. Furthermore, the highest reported yield of HA with high molecular weight for immobilized PmHAS is achieved. This system establishes a foundation for continuous HA formation, with future works potentially enhancing PmHAS stability through protein engineering.
期刊介绍:
Macromolecular Bioscience is a leading journal at the intersection of polymer and materials sciences with life science and medicine. With an Impact Factor of 2.895 (2018 Journal Impact Factor, Journal Citation Reports (Clarivate Analytics, 2019)), it is currently ranked among the top biomaterials and polymer journals.
Macromolecular Bioscience offers an attractive mixture of high-quality Reviews, Feature Articles, Communications, and Full Papers.
With average reviewing times below 30 days, publication times of 2.5 months and listing in all major indices, including Medline, Macromolecular Bioscience is the journal of choice for your best contributions at the intersection of polymer and life sciences.