Noninvasive imaging and quantification of bile salt hydrolase activity: From bacteria to humans

Khodakivskyi, Pavlo; Lauber, Christian L.; Yevtodiyenko, Aleksey; Bazhin, Arkadiy A.; Bruce, Stephen J.; Ringel–Kulka, Tamar; Ringel, Yehuda; Bétrisey, Bertrand; Torres, Joana; Hu, Jianzhong; Chou, Chieh Jason; Goun, Elena A.

doi:10.1126/sciadv.aaz9857

Cited by 35 publications

(27 citation statements)

References 49 publications

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“…Hence, modulating BSH activity is an effective way to impact the composition of the gut ecosystem. Conversely, a damaged BSH activity has been reported in the chronic inflammatory bowel disorder Crohn's disease [61,62], which even led to the recent development of a non-invasive diagnostic tool to assess BSH activity in vivo [63]. Another therapeutic implication was recently demonstrated for recurrent Clostridioides difficile infections that are effectively treated by fecal microbial transplantation (FMT).…”

Section: Discussionmentioning

confidence: 99%

Identifying a Novel Bile Salt Hydrolase from the Keystone Gut Bacterium Christensenella minuta

Déjean¹,

Tudela²,

Bruno³

et al. 2021

Microorganisms

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Christensenella minuta are human gut dwelling bacteria that have been proposed as key members of the gut microbiome, regulating energy balance and adiposity of their host. We formerly identified that a novel strain of C. minuta (strain DSM33407) boosted microbiota diversity and stimulated deconjugation of the primary bile acid taurocholic acid in human samples. However, there is no description of a bile salt hydrolase (BSH) protein carried in the genome of C. minuta. Here, we identified and cloned a protein from C. minuta’s genome that carries a potent BSH activity, which preferentially deconjugates glycine-conjugated bile acids. We then retrieved 14,319 putative BSH sequences from the NCBI database and filtered them using the UHGP database to collect a total of 6701 sequences that were used to build the most comprehensive phylogenetic tree of BSH-related enzymes identified in the human microbiome so far. This phylogenetic tree revealed that C. minuta’s BSH amino acid sequence clusters away from others with a threshold of 70% identity. This is therefore the first description of C. minuta’s BSH protein, which may be involved in its unique role within the human gut microbial ecosystem.

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Section: Discussionmentioning

confidence: 99%

Identifying a Novel Bile Salt Hydrolase from the Keystone Gut Bacterium Christensenella minuta

Déjean¹,

Tudela²,

Bruno³

et al. 2021

Microorganisms

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“…An aminoluciferin-BA conjugated substrate for BSH was recently described. Its hydrolysis efficiency is comparable to that of naturally occurring glycine-conjugated and taurine-conjugated BAs [32].…”

Section: Bsh Bl Probesmentioning

confidence: 82%

“…The use of this method to define the complex microbiota along the entire int tract could complement the recent in vivo imaging approach. Due to the analytic formances in term of linear and dynamic range, it could potentially be used in vivo d colonoscopy intervention [32,46] with minimal invasivity to study the microbio BSH activity along the entire intestinal tract; or it can be used for the quantification o activity in clinical stool samples of patients [32]. The data show a linearity of the light signal at a low enzyme activity (0.2-1.8 U/mL) and the kinetics is faster for the natural substrate GCDCA than for aLuc-CDCA, in line with previous data [33].…”

Section: Bsh Activitymentioning

confidence: 99%

“…As reported in previous papers, the BL probes is highly specific toward BSH in biological samples. In fact, the amide bond of amidated BA is stable in duodenal fluid and bile and it can be hydrolyzed only by BSH enzyme [32,33]. The synthetic BL-probes should only present the requisite of a not hindered N position to allow the formation of amide bond, as it is for aLuc-CDCA, as described in Material and Methods.…”

Section: Bsh Activitymentioning

confidence: 99%

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Compact Miniaturized Bioluminescence Sensor Based on Continuous Air-Segmented Flow for Real-Time Monitoring: Application to Bile Salt Hydrolase (BSH) Activity and ATP Detection in Biological Fluids

et al. 2021

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A simple and versatile continuous air-segmented flow sensor using immobilized luciferase was designed as a general miniaturized platform based on sensitive biochemiluminescence detection. The device uses miniaturized microperistaltic pumps to deliver flows and compact sensitive light imaging detectors based on BI-CMOS (smartphone camera) or CCD technology. The low-cost components and power supply make it suitable as out-lab device at point of need to monitor kinetic-related processes or ex vivo dynamic events. A nylon6 flat spiral carrying immobilized luciferase was placed in front of the detector in lensless mode using a fiber optic tapered faceplate. ATP was measured in samples collected by microdialysis from rat brain with detecting levels as low as 0.4 fmoles. The same immobilized luciferase was also used for the evaluation of bile salt hydrolase (BSH) activity in intestinal microbiota. An aminoluciferin was conjugatated with chenodeoxycholic acid forming the amide derivative aLuc-CDCA. The hydrolysis of the aLuc-CDCA probe by BSH releases free uncaged aminoluciferin which is the active substrate for luciferase leading to light emission. This method can detect as low as 0.5 mM of aLuc-CDCA, so it can be used on real faecal human samples to study BSH activity and its modulation by diseases and drugs.

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“…This approach permitted the differentiation between humanized mice models harboring either a "healthy" or an IBD-affected human microbiome, since the proteins were tagged only in the IBD model and not in the "healthy" one (43). A versatile set of chemical probes for detecting bile salt hydrolase (BSH) activity was based on the bioluminescence of luciferin and has been employed to detect BSH activity in purified enzymes, bacterial cells and fecal slurries (44). They were also used to noninvasively detect BSH activity in mice and humans in the same study.…”

Section: Chemical Biology-based Approaches: Using Chemical Probes As ...mentioning

confidence: 99%

Exploration of Chemical Biology Approaches to Facilitate the Discovery and Development of Novel Antibiotics

2022

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Approximately 2.8 million people worldwide are infected with bacteria that are deemed resistant to clinically relevant antibiotics. This accounts for 700,000 deaths every year and represents a major public health threat that has been on the rise for the past two decades. In contrast, the pace of antibiotic discovery to treat these resistant pathogens has significantly decreased. Most antibiotics are complex natural products that were isolated from soil microorganisms during the golden era of antibiotic discovery (1940s to 1960s) employing the “Waksman platform”. After the collapse of this discovery platform, other strategies and approaches emerged, including phenotype- or target-based screenings of large synthetic compound libraries. However, these methods have not resulted in the discovery and/or development of new drugs for clinical use in over 30 years. A better understanding of the structure and function of the molecular components that constitute the bacterial system is of paramount importance to design new strategies to tackle drug-resistant pathogens. Herein, we review the traditional approaches as well as novel strategies to facilitate antibiotic discovery that are chemical biology-focused. These include the design and application of chemical probes that can undergo bioorthogonal reactions, such as copper (I)-catalyzed azide-alkyne cycloadditions (CuAAC). By specifically interacting with bacterial proteins or being incorporated in the microorganism’s metabolism, chemical probes are powerful tools in drug discovery that can help uncover new drug targets and investigate the mechanisms of action and resistance of new antibacterial leads.

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Noninvasive imaging and quantification of bile salt hydrolase activity: From bacteria to humans

Cited by 35 publications

References 49 publications

Identifying a Novel Bile Salt Hydrolase from the Keystone Gut Bacterium Christensenella minuta

Identifying a Novel Bile Salt Hydrolase from the Keystone Gut Bacterium Christensenella minuta

Compact Miniaturized Bioluminescence Sensor Based on Continuous Air-Segmented Flow for Real-Time Monitoring: Application to Bile Salt Hydrolase (BSH) Activity and ATP Detection in Biological Fluids

Exploration of Chemical Biology Approaches to Facilitate the Discovery and Development of Novel Antibiotics

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