Optimized 3'-Phosphoadenosine-5'-phosphosulfate Biosynthesis via Yeast-Powered ATP Regeneration and Biotin-Streptavidin Enzyme Immobilization.

IF 3 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry Biochemistry Pub Date : 2025-09-18 DOI:10.1021/acs.biochem.5c00373

Xin-Yu Li, Jian-Qun Deng, Yu-Han Zhao, Ya-Lin Cao, Yi Li, Jin Hou, Xue-Ping Guo, Ju-Zheng Sheng

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引用次数: 0

Abstract

3'-Phosphoadenosine-5'-phosphosulfate (PAPS), a universal sulfate donor for sulfation reactions, is indispensable for synthesizing bioactive molecules including therapeutic glycosaminoglycans and sulfolipids; however, its enzymatic production on an industrial scale is constrained by ATP overconsumption and the limited free enzyme reusability. We report an integrated biocatalytic platform combining ATP regeneration with affinity immobilization to enable sustainable PAPS biosynthesis. A polyphosphate kinase-driven ATP regeneration system achieved 86% PAPS conversion efficiency by regenerating ADP using low-cost polyphosphate. Biotin-streptavidin affinity immobilization enhanced operational stability, retaining >50% activity over six reuse cycles with a cumulative PAPS titer of 12.02 g/L. Coupling adenosine-converting Saccharomyces cerevisiae whole-cell catalysts with this system decreased substrate costs by 80.7% and delivered 96% molar PAPS yield from adenosine. This work provides a sustainable platform for industrial PAPS biosynthesis to promote sulfated biomolecule production, including glycosaminoglycans and other therapeutics.

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酵母驱动ATP再生和生物素-链亲和素酶固定化优化3'-磷酸腺苷-5'-硫酸磷酸生物合成。

3′-磷酸腺苷-5′-磷酸硫酸酯（PAPS）是磺化反应的普遍硫酸供体，是合成治疗性糖胺聚糖和磺化脂类等生物活性分子不可或缺的物质；然而，在工业规模上，它的酶生产受到ATP过度消耗和有限的游离酶可重用性的限制。我们报道了一个集成的生物催化平台，结合ATP再生和亲和固定，以实现可持续的PAPS生物合成。利用低成本的聚磷酸盐再生ADP，一种多磷酸激酶驱动的ATP再生系统实现了86%的PAPS转化效率。生物素-链亲和素亲和固定提高了操作稳定性，在6个重复使用周期内保持了50%的活性，累积PAPS滴度为12.02 g/L。将腺苷转化酿酒酵母全细胞催化剂与该体系偶联，使底物成本降低80.7%，腺苷的摩尔PAPS产率达到96%。这项工作为工业PAPS生物合成提供了一个可持续的平台，以促进硫酸生物分子的生产，包括糖胺聚糖和其他治疗药物。

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来源期刊

Biochemistry Biochemistry 生物-生化与分子生物学

CiteScore

5.50

自引率

3.40%

发文量

336

审稿时长

1-2 weeks

期刊介绍： Biochemistry provides an international forum for publishing exceptional, rigorous, high-impact research across all of biological chemistry. This broad scope includes studies on the chemical, physical, mechanistic, and/or structural basis of biological or cell function, and encompasses the fields of chemical biology, synthetic biology, disease biology, cell biology, nucleic acid biology, neuroscience, structural biology, and biophysics. In addition to traditional Research Articles, Biochemistry also publishes Communications, Viewpoints, and Perspectives, as well as From the Bench articles that report new methods of particular interest to the biological chemistry community.