微生物组中聚糖转化的活性跟踪研究。

IF 14.4 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY
Conor J Crawford, Greta Reintjes, Vipul Solanki, Manuel G Ricardo, Jens Harder, Rudolf Amann, Jan-Hendrik Hehemann, Peter H Seeberger
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引用次数: 0

摘要

聚糖塑造海洋和肠道中的微生物群,推动全球碳循环和人类健康的关键步骤。然而,我们在微生物组中追踪微生物聚糖周转的能力是有限的,因为在没有事先基因组知识的情况下识别活性降解物仍然是一个关键的挑战。在这里,我们介绍了一种基于活性的荧光共振能量转移(FRET)探针,可以直接可视化和量化复杂微生物群落中的聚糖代谢。为了证明这一概念,我们研究了α-甘露聚糖的降解,这是一种在藻华中突出的多糖。利用自动聚糖组装,我们合成了一种带有荧光素-罗丹明FRET对的甘露聚糖六糖。利用Salegentibacter sp. Hel_I_6重组表达的内切-α-甘露聚糖酶(GH76)对探针进行了验证。它被证明在细胞裂解物,纯培养物和复杂微生物组中起作用(通过平板实验和显微镜)。该探针实现了海洋微生物群中α-甘露聚糖原位转换的时空可视化。聚糖FRET探针是多功能的工具,用于跟踪从单一酶到微生物组的生物尺度上的聚糖降解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Activity-Based Tracking of Glycan Turnover in Microbiomes.

Glycans shape microbiomes in the ocean and the gut, driving key steps in the global carbon cycle and human health. Yet, our ability to track microbial glycan turnover across microbiomes is limited, as identifying active degraders without prior genomic knowledge remains a key challenge. Here, we introduce an activity-based fluorescence resonance energy transfer (FRET) probe that enables direct visualization and quantification of glycan metabolism in complex microbial communities. As a proof of concept, we investigated α-mannan degradation, a prominent polysaccharide in algal blooms. Using automated glycan assembly, we synthesized a mannan hexasaccharide bearing a fluorescein-rhodamine FRET pair. The probe was validated using a recombinantly expressed endo-α-mannanase (GH76) from Salegentibacter sp. Hel_I_6. It was shown to function in cell lysates, pure cultures, and complex microbiomes (via plate assays and microscopy). This probe enabled spatiotemporal visualization of in situ α-mannan turnover in a marine microbiome. Glycan FRET probes are versatile tools for tracking glycan degradation across biological scales from single enzymes to microbiomes.

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来源期刊
CiteScore
24.40
自引率
6.00%
发文量
2398
审稿时长
1.6 months
期刊介绍: The flagship journal of the American Chemical Society, known as the Journal of the American Chemical Society (JACS), has been a prestigious publication since its establishment in 1879. It holds a preeminent position in the field of chemistry and related interdisciplinary sciences. JACS is committed to disseminating cutting-edge research papers, covering a wide range of topics, and encompasses approximately 19,000 pages of Articles, Communications, and Perspectives annually. With a weekly publication frequency, JACS plays a vital role in advancing the field of chemistry by providing essential research.
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