Dataset for "A Function-Driven Single-Cell Approach to Unveiling and Cultivating Hidden Microplastic-Degrading Microbes from Insect Gut Microbiota"

Biodegradation is a sustainable strategy to address global microplastics (MPs) pollution but is constrained by the lack of efficient degrading microbes and effective tools to harness them. Here, we developed a function-driven single-cell approach to precisely identify and recover MPs-degrading microorganisms from complex microbiota by integrating isotope-labeled single-cell Raman spectroscopy with targeted cell sorting, sequencing and culturing. Using heavy water and MPs as the sole carbon source, Raman spectroscopy effectively identified active microbes capable of degrading multiple types of MPs from insect gut microbiota. Raman-guided single-cell sorting and sequencing revealed seven previously unrecognized polystyrene degraders and mapped key enzymes involved in each degradation stage. Furthermore, live-cell Raman-guided sorting enabled the cultivation of rare but highly active polystyrene degraders often missed by conventional methods. This “screen-first, culture-second” single-cell approach offers a powerful and scalable platform to accelerate MPs biodegradation and supports the development of microbial solutions to mitigate global plastic pollution.

生物降解（Biodegradation）是应对全球微塑料（microplastics, MPs）污染的可持续策略，但受限于缺乏高效降解微生物及利用这些微生物的有效工具。在此，我们开发了一种功能驱动的单细胞方法，通过整合同位素标记单细胞拉曼光谱（isotope-labeled single-cell Raman spectroscopy）、靶向细胞分选、测序及培养技术，从复杂微生物群落中精准识别并回收微塑料降解微生物。以重水和微塑料为唯一碳源，拉曼光谱可有效识别昆虫肠道微生物群落中能降解多种微塑料的活性微生物。拉曼引导的单细胞分选与测序揭示了7种此前未被发现的聚苯乙烯降解菌，并绘制了各降解阶段涉及的关键酶图谱。此外，活细胞拉曼引导的分选技术能够培养出常规方法常遗漏的稀有但高活性聚苯乙烯降解菌。这种“先筛选、后培养”的单细胞方法提供了一个强大且可扩展的平台，以加速微塑料生物降解，并助力开发微生物解决方案以缓解全球塑料污染。