Hyaluronan (HA) is a linear polysaccharide composed of repeating disaccharide units. It has been well documented to play an array of biological functions in cancer events. Here, we reported a sequential one-pot multienzyme (OPME) strategy for in vitro synthesis of HA and its derivatives. The strategy, which combined in situ sugar nucleotides generation with HA chain polymerization, could convert cheap monosaccharides into HA polymers without consuming exogenous sugar nucleotide donors. HA polymers (number-average molecular weight ranged from 1.5 × 10⁴ to 5.5 × 10⁵ Da) with over 70% yields were efficiently synthesized and purified from this one-pot system. More importantly, partial labeled HA derivative was further synthesized by metabolic incorporation of unnatural monosaccharide analogues into the sequential OPME system. Cross-linked HA hydrogel was achieved via copper (I)-catalyzed azide-alkyne cycloaddition and exhibited novel networks consisting of both inter- and intra-connected HA chains, which could facilitate the potential applications of this unique polysaccharide.

中文翻译：

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透明质酸及其衍生物的顺序一锅多酶合成

透明质酸（HA）是由重复的二糖单元组成的线性多糖。它已被充分证明在癌症事件中起着一系列生物学功能。在这里，我们报告了HA及其衍生物的体外合成的顺序一锅多酶（OPME）策略。该策略将原位糖核苷酸生成与HA链聚合相结合，可以在不消耗外源糖核苷酸供体的​​情况下将廉价的单糖转变为HA聚合物。HA聚合物（数均分子量范围为1.5×10 ⁴至5.5×10 ⁵ 从此一锅系统中高效合成并纯化了收率超过70％的Da）。更重要的是，通过将非天然单糖类似物代谢并入顺序的OPME系统中，进一步合成了部分标记的HA衍生物。交联的HA水凝胶是通过铜（I）催化的叠氮化物-炔烃环加成反应获得的，并展现出由相互连接的HA链和内部连接的HA链组成的新型网络，这可以促进这种独特多糖的潜在应用。