Synthesis and evaluation of a series of 6-chloro-4-methylumbelliferyl glycosides as fluorogenic reagents for screening metagenomic libraries for glycosidase activity

Carbohydrate Research
Hong-Ming ChenStephen G Withers

Abstract

Screening of large enzyme libraries such as those derived from metagenomic sources requires sensitive substrates. Fluorogenic glycosides typically offer the best sensitivity but typically must be used in a stopped format to generate good signal. Use of fluorescent phenols of pKa < 7, such as halogenated coumarins, allows direct screening at neutral pH. The synthesis and characterisation of a set of nine different glycosides of 6-chloro-4-methylumbelliferone are described. The use of these substrates in a pooled format for screening of expressed metagenomic libraries yielded a "hit rate" of 1 in 60. Hits were then readily deconvoluted with the individual substrates in a single plate to identify specific activities within each clone. The use of such a collection of substrates greatly accelerates the screening process.

References

Dec 23, 2006·Trends in Biotechnology·David J Pollard, John M Woodley
Jan 30, 2007·Organic Letters·Sungjin Park, Injae Shin
Sep 4, 2008·Organic Letters·Jean-Alexandre RichardAnthony Romieu
Mar 29, 2011·Current Opinion in Biotechnology·Marcus TauppSteven J Hallam
Oct 29, 2011·Chemical Society Reviews·Roger A Sheldon
Jun 29, 2012·Bioscience, Biotechnology, and Biochemistry·Liangbin ZhouRongmin Yu
Jan 17, 2013·Bioorganic & Medicinal Chemistry·Hao-Wen KaoHsin-Ell Wang
Aug 3, 2013·Journal of Biotechnology·Keith MewisSteven J Hallam
Jun 10, 2014·Annual Review of Chemical and Biomolecular Engineering·Nidhi Tibrewal, Yi Tang
Mar 24, 2015·Frontiers in Bioengineering and Biotechnology·Susan Anne BaldwinHamed Sanei
Aug 2, 2015·Current Opinion in Chemical Biology·Zachary ArmstrongSteven J Hallam

❮ Previous
Next ❯

Citations

Sep 30, 2017·Chembiochem : a European Journal of Chemical Biology·Tianyuan PengNicola L B Pohl
Jul 13, 2018·Angewandte Chemie·Seyed Amirhossein NasseriStephen G Withers
Oct 6, 2017·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Hirak SaxenaWarren Wakarchuk
Sep 20, 2020·Applied and Environmental Microbiology·Maria João Maurício da FonsecaYves Briers

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

American Biotechnology Laboratory
E Koller
American Journal of Hematology
Andreas Greinacher
Methods in Molecular Biology
A P Corfield, N Myerscough
Trends in Biochemical Sciences
L Holm, C Sander
© 2022 Meta ULC. All rights reserved