Protein engineering study of beta-mannosidase to set up a
potential chemically efficient biocatalyst

J 2014

Protein engineering study of beta-mannosidase to set up a potential chemically efficient biocatalyst

DEMO, Gabriel; Veronika HORSKÁ; Barbora FLIEDROVA; Jakub ŠTĚPÁN; Jaroslav KOČA et al.

Základní údaje

Originální název

Protein engineering study of beta-mannosidase to set up a potential chemically efficient biocatalyst

Autoři

DEMO, Gabriel; Veronika HORSKÁ; Barbora FLIEDROVA; Jakub ŠTĚPÁN; Jaroslav KOČA; Lenka WEIGNEROVA; Vladimír KŘEN a Michaela WIMMEROVÁ

Vydání

Glycobiology, Oxford, Oxford University Press, 2014, 0959-6658

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.147

Kód RIV

RIV/00216224:14740/14:00073772

Organizační jednotka

Středoevropský technologický institut

DOI

https://doi.org/10.1093/glycob/cwu074

UT WoS

000347410300011

EID Scopus

2-s2.0-84940761366

Klíčová slova česky

dokovani beta-mannosidasa molekulova dynamika mutagenese proteinove inzenirstvi

Klíčová slova anglicky

Docking beta-mannosidase molecular dynamics mutagenesis protein engineering

Štítky

kontrola MP, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 10. 3. 2015 16:40, Martina Prášilová

Anotace

V originále

This study is focused on the analysis and mutagenesis of beta-mannosidase from Bacteroides thetaiotaomicron with the aim of broadening its substrate specificity to 2-acetamido-2-deoxy-beta-d-mannopyranosyl (beta-ManNAc) derivatives. Various conformations (4C1, 4H5, and 1S5) of native and modified ligands were docked to the binding site of the protein to determine the most suitable conformation of sugars for further hydrolysis. Key amino acid residues were mutated in silico focusing on stabilizing the acetamido group of beta-ManNAc as well as forming the oxazoline intermediate needed for hydrolysis. The results of large set of 5 ns molecular dynamic simulations showed that the majority of the active site residues are involved in substrate interaction and do not exhibit a higher flexibility except for Asn178. Mutations of Asn178 to alanine and Asp199 to serine could lead to a stabilisation of the acetamido group in the binding site. So far, in vitro mutagenesis and the screen of a large variety of biological sources were unable to extend beta-mannosidase's activity to include beta-ManNAc derivatives.

Návaznosti

ED1.1.00/02.0068, projekt VaV

Název: CEITEC - central european institute of technology

GAP207/10/0321, projekt VaV

Název: Enzymy metabolismu mannosidů v přípravě N-acetyl-mannosaminových struktur

Investor: Grantová agentura ČR, Enzymy metabolismu mannosidů v přípravě N-acetyl-mannosaminových struktur

Citovat

DEMO, Gabriel; Veronika HORSKÁ; Barbora FLIEDROVA; Jakub ŠTĚPÁN; Jaroslav KOČA; Lenka WEIGNEROVA; Vladimír KŘEN a Michaela WIMMEROVÁ. Protein engineering study of beta-mannosidase to set up a potential chemically efficient biocatalyst. Glycobiology. Oxford: Oxford University Press, 2014, roč. 24, č. 12, s. 1301-1311. ISSN 0959-6658. Dostupné z: https://doi.org/10.1093/glycob/cwu074.

@article{1194254,
   author = {Demo, Gabriel and Horská, Veronika and Fliedrova, Barbora and Štěpán, Jakub and Koča, Jaroslav and Weignerova, Lenka and Křen, Vladimír and Wimmerová, Michaela},
   article_location = {Oxford},
   article_number = {12},
   doi = {https://doi.org/10.1093/glycob/cwu074},
   keywords = {Docking beta-mannosidase molecular dynamics mutagenesis protein engineering},
   language = {eng},
   issn = {0959-6658},
   journal = {Glycobiology},
   title = {Protein engineering study of beta-mannosidase to set up a potential chemically efficient biocatalyst},
   url = {http://glycob.oxfordjournals.org/content/24/12/1301.long},
   volume = {24},
   year = {2014}
}

TY  - JOUR
ID  - 1194254
AU  - Demo, Gabriel - Horská, Veronika - Fliedrova, Barbora - Štěpán, Jakub - Koča, Jaroslav - Weignerova, Lenka - Křen, Vladimír - Wimmerová, Michaela
PY  - 2014
TI  - Protein engineering study of beta-mannosidase to set up a potential chemically efficient biocatalyst
JF  - Glycobiology
VL  - 24
IS  - 12
SP  - 1301-1311
EP  - 1301-1311
PB  - Oxford University Press
SN  - 09596658
KW  - Docking beta-mannosidase molecular dynamics mutagenesis protein engineering
UR  - http://glycob.oxfordjournals.org/content/24/12/1301.long
L2  - http://glycob.oxfordjournals.org/content/24/12/1301.long
N2  - This study is focused on the analysis and mutagenesis of beta-mannosidase from Bacteroides thetaiotaomicron with the aim of broadening its substrate specificity to 2-acetamido-2-deoxy-beta-d-mannopyranosyl (beta-ManNAc) derivatives. Various conformations (4C1, 4H5, and 1S5) of native and modified ligands were docked to the binding site of the protein to determine the most suitable conformation of sugars for further hydrolysis. Key amino acid residues were mutated in silico focusing on stabilizing the acetamido group of beta-ManNAc as well as forming the oxazoline intermediate needed for hydrolysis. The results of large set of 5 ns molecular dynamic simulations showed that the majority of the active site residues are involved in substrate interaction and do not exhibit a higher flexibility except for Asn178. Mutations of Asn178 to alanine and Asp199 to serine could lead to a stabilisation of the acetamido group in the binding site. So far, in vitro mutagenesis and the screen of a large variety of biological sources were unable to extend beta-mannosidase's activity to include beta-ManNAc derivatives.
ER  -

DEMO, Gabriel; Veronika HORSKÁ; Barbora FLIEDROVA; Jakub ŠTĚPÁN; Jaroslav KOČA; Lenka WEIGNEROVA; Vladimír KŘEN a Michaela WIMMEROVÁ. Protein engineering study of beta-mannosidase to set up a potential chemically efficient biocatalyst. \textit{Glycobiology}. Oxford: Oxford University Press, 2014, roč.~24, č.~12, s.~1301-1311. ISSN~0959-6658. Dostupné z: https://doi.org/10.1093/glycob/cwu074.

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