Profiling rhythmicity of bile salt hydrolase activity in the gut lumen with a rapid fluorescence assay.
Profiling rhythmicity of bile salt hydrolase activity in the gut lumen with a rapid fluorescence assay.
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用快速的荧光测定法中胆汁盐水解酶活性的概况分析。
DOI：
10.1039/d2ob02257e
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发表时间：
2023-05-17
影响因子：
3.2
通讯作者：
中科院分区：
文献类型：
--
作者：
细胞功能的昼夜节律性是地球上大多数生物生存的关键。许多昼夜节律功能是由大脑驱动的，但对一套单独的外周节律的调节仍然知之甚少。肠道微生物组是调节宿主外周节律的潜在候选者，本研究试图具体研究微生物胆盐的生物转化过程。为了能够进行这项工作，有必要对胆盐水解酶(BSH)进行一种可以用于少量粪便样本的检测。使用开启的荧光探针，我们开发了一种快速且廉价的方法来检测bsh酶的活性，其浓度低至6-25μM，比以前的方法更可靠。我们成功地应用这种基于罗丹明的方法检测了多种生物样本中的BSH活性，如重组蛋白、全细胞、粪便样本和小鼠肠道内容物。我们能够在2小时内从少量的小鼠粪便/肠道内容物(20-50毫克)中检测到显著的BSH活性，这表明它在各种生物/临床应用中的潜力。用该方法研究了小鼠大肠BSH活性的昼夜变化。利用时间限制的摄食条件，我们提供了微生物组BSH活性水平24小时节律性的直接证据，并表明这种节律性受摄食方式的影响。我们新的以功能为中心的方法有可能帮助发现治疗、饮食或生活方式干预，以纠正与胆汁代谢有关的昼夜节律扰动。胆盐水解酶活性水平的24小时节律性受摄食方式的影响，为肠道微生物群影响宿主外周昼夜节律性提供了一条途径。
Diurnal rhythmicity of cellular function is key to survival for most organisms on Earth. Many circadian functions are driven by the brain, but regulation of a separate set of peripheral rhythms remains poorly understood. The gut microbiome is a potential candidate for regulation of host peripheral rhythms, and this study sought to specifically examine the process of microbial bile salt biotransformation. To enable this work, an assay for bile salt hydrolase (BSH) that could work with small quantities of stool samples was necessary. Using a turn-on fluorescence probe, we developed a rapid and inexpensive assay to detect BSH enzyme activity with concentrations as low as 6–25 μM, which is considerably more robust than prior approaches. We successfully applied this rhodamine-based assay to detect BSH activity in a wide range of biological samples such as recombinant protein, whole cells, fecal samples, and gut lumen content from mice. We were able to detect significant BSH activity in small amounts of mouse fecal/gut content (20–50 mg) within 2 h, which illustrates its potential for use in various biological/clinical applications. Using this assay, we investigated the diurnal fluctuations of BSH activity in the large intestine of mice. By using time restricted feeding conditions, we provided direct evidence of 24 h rhythmicity in microbiome BSH activity levels and showed that this rhythmicity is influenced by feeding patterns. Our novel function-centric approach has potential to aid in the discovery of therapeutic, diet, or lifestyle interventions for correction of circadian perturbations linked to bile metabolism. 24-hour rhythmicity in bile salt hydrolase activity levels is influenced by feeding patterns, providing a pathway for the gut microbiome to affect host peripheral circadian rhythmicity.
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胆汁盐水解酶活性的无创成像和定量：从细菌到人。
影响因子：
13.6
作者：
Khodakivskyi PV;Lauber CL;Yevtodiyenko A;Bazhin AA;Bruce S;Ringel-Kulka T;Ringel Y;Bétrisey B;Torres J;Hu J;Chou CJ;Goun EA
通讯作者：
Goun EA
影响因子：
5.1
作者：
Flynn-Evans EE;Barger LK;Kubey AA;Sullivan JP;Czeisler CA
通讯作者：
Czeisler CA
DOI：
10.3109/09540261.2014.911149
发表时间：
2014-04
期刊：
International review of psychiatry (Abingdon, England)
影响因子：
--
作者：
Baron KG;Reid KJ
通讯作者：
Reid KJ
影响因子：
4
作者：
Adhikari, Arijit A.;Ramachandran, Deepti;Chaudhari, Snehal N.;Powell, Chelsea E.;Li, Wei;McCurry, Megan D.;Banks, Alexander S.;Devlin, A. Sloan
通讯作者：
Devlin, A. Sloan
影响因子：
4.6
作者：
Chellappa SL;Morris CJ;Scheer FAJL
通讯作者：
Scheer FAJL