High‐Throughput Colorimetric Detection and Quantification of Indoles and Pyrroloindoles for Enzymatic Activity Determination DOI Creative Commons
Diana A. Amariei, Mona Haase, Moritz K. T. Klischan

и другие.

ChemCatChem, Год журнала: 2024, Номер 16(11)

Опубликована: Фев. 20, 2024

Abstract Indoles and pyrroloindoles are structural motifs present in many biologically active natural products. Multiple classes of enzymes catalyze the transformation indoles into via group transfer followed by intramolecular cyclization, such as peroxydases, methyltransferases, prenyltransferases. Due to selective introduction a stereogenic center, these receive increasing attention catalytic tools for production pharmacologically relevant compounds. Two new colorimetric assays described this work, which allow quantification enzymatic reactions from perspective substrate product. For substrates, indole assay is based on modified version Ehrlich test, with use light driving force color formation. The pyrroloindole uses cerium sulfate reagent complementary both were successfully utilized activity determination C3‐indole methyltransferase. They can facilitate high‐throughput screening mutant libraries, offering support engineering enzymes, but also be used stand‐alone methods detection

Язык: Английский

Redox regulation: mechanisms, biology and therapeutic targets in diseases DOI Creative Commons
Bowen Li, Hui Ming, Siyuan Qin

и другие.

Signal Transduction and Targeted Therapy, Год журнала: 2025, Номер 10(1)

Опубликована: Март 7, 2025

Redox signaling acts as a critical mediator in the dynamic interactions between organisms and their external environment, profoundly influencing both onset progression of various diseases. Under physiological conditions, oxidative free radicals generated by mitochondrial respiratory chain, endoplasmic reticulum, NADPH oxidases can be effectively neutralized NRF2-mediated antioxidant responses. These responses elevate synthesis superoxide dismutase (SOD), catalase, well key molecules like nicotinamide adenine dinucleotide phosphate (NADPH) glutathione (GSH), thereby maintaining cellular redox homeostasis. Disruption this finely tuned equilibrium is closely linked to pathogenesis wide range Recent advances have broadened our understanding molecular mechanisms underpinning dysregulation, highlighting pivotal roles genomic instability, epigenetic modifications, protein degradation, metabolic reprogramming. findings provide foundation for exploring regulation mechanistic basis improving therapeutic strategies. While antioxidant-based therapies shown early promise conditions where stress plays primary pathological role, efficacy diseases characterized complex, multifactorial etiologies remains controversial. A deeper, context-specific signaling, particularly redox-sensitive proteins, designing targeted aimed at re-establishing balance. Emerging small molecule inhibitors that target specific cysteine residues proteins demonstrated promising preclinical outcomes, setting stage forthcoming clinical trials. In review, we summarize current intricate relationship disease also discuss how these insights leveraged optimize strategies practice.

Язык: Английский

Процитировано

5

Ferulic acid production in Escherichia coli by engineering caffeic acid O-methyltransferase DOI

Di Qiu,

Haifeng Ding, Chao Zhou

и другие.

International Journal of Biological Macromolecules, Год журнала: 2025, Номер 306, С. 141764 - 141764

Опубликована: Март 4, 2025

Язык: Английский

Процитировано

0

Fluorogenic, Subsingle-Turnover Monitoring of Enzymatic Reactions Involving NAD(P)H Provides a Generalized Platform for Directed Ultrahigh-Throughput Evolution of Biocatalysts in Microdroplets DOI Creative Commons
Matthew Penner, Oskar James Klein, Maximilian Gantz

и другие.

Journal of the American Chemical Society, Год журнала: 2025, Номер unknown

Опубликована: Март 24, 2025

Enzyme engineering and discovery are crucial for a sustainable future bioeconomy. Harvesting new biocatalysts from large libraries through directed evolution or functional metagenomics requires accessible, rapid assays. Ultrahigh-throughput screening formats often require optical readouts, leading to the use of model substrates that may misreport target activity necessitate bespoke synthesis. This is particular challenge when glycosyl hydrolases, which leverage molecular recognition beyond glycosidic bond, so complex chemical synthesis would have be deployed build fluoro- chromogenic substrate. In contrast, coupled assays represent modular "plug-and-play" system: any enzyme–substrate pairing can investigated, provided reaction produce common intermediate links catalytic detection cascade readout. Here, we establish producing fluorescent readout in response NAD(P)H via glutathione reductase subsequent thiol-mediated uncaging reaction, with low nanomolar limit plates. Further scaling down microfluidic droplet possible: fluorophore leakage-free report 3 orders magnitude-improved sensitivity compared absorbance-based systems, resolution 361,000 product molecules per droplet. Our approach enables nonfluorogenic droplet-based enrichments, applicability hydrolases imine reductases (IREDs). To demonstrate assay's readiness combinatorial experiments, one round was performed select glycosidase processing natural substrate, beechwood xylan, improved kinetic parameters pool >106 mutagenized sequences.

Язык: Английский

Процитировано

0

Quantitative detection and cellular imaging of hydrogen sulfide using a SERS probe based on AuAg nanocages DOI
Xiaoli Wang, Jiale Zhang, Yun Wang

и другие.

Biosensors and Bioelectronics, Год журнала: 2025, Номер unknown, С. 117580 - 117580

Опубликована: Май 1, 2025

Язык: Английский

Процитировано

0

High‐Throughput Colorimetric Detection and Quantification of Indoles and Pyrroloindoles for Enzymatic Activity Determination DOI Creative Commons
Diana A. Amariei, Mona Haase, Moritz K. T. Klischan

и другие.

ChemCatChem, Год журнала: 2024, Номер 16(11)

Опубликована: Фев. 20, 2024

Abstract Indoles and pyrroloindoles are structural motifs present in many biologically active natural products. Multiple classes of enzymes catalyze the transformation indoles into via group transfer followed by intramolecular cyclization, such as peroxydases, methyltransferases, prenyltransferases. Due to selective introduction a stereogenic center, these receive increasing attention catalytic tools for production pharmacologically relevant compounds. Two new colorimetric assays described this work, which allow quantification enzymatic reactions from perspective substrate product. For substrates, indole assay is based on modified version Ehrlich test, with use light driving force color formation. The pyrroloindole uses cerium sulfate reagent complementary both were successfully utilized activity determination C3‐indole methyltransferase. They can facilitate high‐throughput screening mutant libraries, offering support engineering enzymes, but also be used stand‐alone methods detection

Язык: Английский

Процитировано

1