Novel α-L-Fucosidases from a Soil Metagenome for Production of Fucosylated Human Milk Oligosaccharides

PloS One
Mateusz LezykJørn D Mikkelsen

Abstract

This paper describes the discovery of novel α-L-fucosidases and evaluation of their potential to catalyse the transglycosylation reaction leading to production of fucosylated human milk oligosaccharides. Seven novel α-L-fucosidase-encoding genes were identified by functional screening of a soil-derived metagenome library and expressed in E. coli as recombinant 6xHis-tagged proteins. All seven fucosidases belong to glycosyl hydrolase family 29 (GH 29). Six of the seven α-L-fucosidases were substrate-inhibited, moderately thermostable and most hydrolytically active in the pH range 6-7, when tested with para-nitrophenyl-α-L-fucopyranoside (pNP-Fuc) as the substrate. In contrast, one fucosidase (Mfuc6) exhibited a high pH optimum and an unusual sigmoidal kinetics towards pNP-Fuc substrate. When tested for trans-fucosylation activity using pNP-Fuc as donor, most of the enzymes were able to transfer fucose to pNP-Fuc (self-condensation) or to lactose. With the α-L-fucosidase from Thermotoga maritima and the metagenome-derived Mfuc5, different fucosyllactose variants including the principal fucosylated HMO 2'-fucosyllactose were synthesised in yields of up to ~6.4%. Mfuc5 was able to release fucose from xyloglucan and could also use i...Continue Reading

References

Sep 1, 1997·Nucleic Acids Research·S F AltschulD J Lipman
Mar 26, 2003·Glycobiology·Daniel J Becker, John B Lowe
Sep 3, 2004·The Journal of Pediatrics·Ardythe L MorrowDavid S Newburg
Jun 15, 2007·The Plant Journal : for Cell and Molecular Biology·Isabel MollerWilliam G T Willats
Dec 1, 2009·Biotechnology Journal·Marla TuffinDon A Cowan
Nov 27, 2010·Nucleic Acids Research·Aron Marchler-BauerStephen H Bryant
Dec 21, 2010·Applied and Environmental Microbiology·Carola Simon, Rolf Daniel
Nov 29, 2011·Biotechnology Advances·Nam Soo HanJin-Ho Seo
Dec 25, 2012·Genome Biology·Sébastien BoisvertJacques Corbeil
Jan 25, 2013·Journal of Agricultural and Food Chemistry·Hassan Ahmadi GavlighiJ Dalgaard Mikkelsen
Apr 2, 2013·Applied and Environmental Microbiology·Jesús Rodríguez-DíazMaría J Yebra
Oct 15, 2013·Genomics & Informatics·Myung Hwan Lee, Seon-Woo Lee
Oct 18, 2013·Molecular Biology and Evolution·Koichiro TamuraSudhir Kumar
Aug 3, 2014·The Journal of Biological Chemistry·Hongnan CaoGeorge N Phillips
Sep 11, 2014·Journal of Agricultural and Food Chemistry·Birgitte ZeunerAnne S Meyer

❮ Previous
Next ❯

Citations

Sep 12, 2020·Critical Reviews in Food Science and Nutrition·Emmanuel Pérez-EscalanteLuis Guillermo González-Olivares
Oct 14, 2017·FEMS Microbiology Letters·Francesca BeriniFlavia Marinelli
Apr 26, 2020·Cellular and Molecular Life Sciences : CMLS·Haiyang WuNathalie Juge
Dec 7, 2018·Biotechnology and Applied Biochemistry·Francisco Guzmán-RodríguezAlma Cruz-Guerrero
Dec 12, 2018·Zeitschrift Für Naturforschung. C, a Journal of Biosciences·Jan Muschiol, Anne S Meyer
May 3, 2018·Applied Biochemistry and Biotechnology·Francisco Guzmán-RodríguezAlma Cruz-Guerrero
May 2, 2020·Applied Microbiology and Biotechnology·Li WanWanmeng Mu
Mar 31, 2021·Biotechnology Advances·Zuzana MészárosKristýna Slámová
May 4, 2021·Extremophiles : Life Under Extreme Conditions·Francisco Guzmán-RodríguezAlma Cruz-Guerrero
May 4, 2021·Critical Reviews in Food Science and Nutrition·Yingying ZhuWanmeng Mu
Jun 19, 2021·Communications Biology·Maria Dalgaard MikkelsenPeter Ulvskov
Jul 28, 2021·Applied Biochemistry and Biotechnology·Mónica A Robles-AriasAlma E Cruz-Guerrero
Aug 28, 2021·International Journal of Molecular Sciences·José Antonio CurielJuan Luis Arqués
Sep 1, 2021·Biotechnology Advances·Zuzana MészárosKristýna Slámová

❮ Previous
Next ❯

Datasets Mentioned

BETA
KJ626336
KJ626337
KJ626338
KJ626339
KJ626340
KJ626341
KJ626342

Methods Mentioned

BETA
electrophoresis
protein assay
nuclear magnetic resonance
NMR
reverse phase chromatography

Software Mentioned

CAZYmes Analysis Toolkit
NCBI Conserved Domain
BLAST P
GeneArt AG
BLAST
Ray Meta
GraphPad Prism
MEGA6

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.