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.

Basic information

Original name

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

Authors

DEMO, Gabriel (703 Slovakia, belonging to the institution), Veronika HORSKÁ (203 Czech Republic, belonging to the institution), Barbora FLIEDROVA (203 Czech Republic), Jakub ŠTĚPÁN (203 Czech Republic, belonging to the institution), Jaroslav KOČA (203 Czech Republic, belonging to the institution), Lenka WEIGNEROVA (203 Czech Republic), Vladimír KŘEN (203 Czech Republic) and Michaela WIMMEROVÁ (203 Czech Republic, guarantor, belonging to the institution)

Edition

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

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 3.147

RIV identification code

RIV/00216224:14740/14:00073772

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1093/glycob/cwu074

UT WoS

000347410300011

Keywords (in Czech)

dokovani beta-mannosidasa molekulova dynamika mutagenese proteinove inzenirstvi

Keywords in English

Docking beta-mannosidase molecular dynamics mutagenesis protein engineering

Abstract

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.

Links

ED1.1.00/02.0068, research and development project

Name: CEITEC - central european institute of technology

GAP207/10/0321, research and development project

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

Investor: Czech Science Foundation

Citovat

DEMO, Gabriel, Veronika HORSKÁ, Barbora FLIEDROVA, Jakub ŠTĚPÁN, Jaroslav KOČA, Lenka WEIGNEROVA, Vladimír KŘEN and Michaela WIMMEROVÁ. Protein engineering study of beta-mannosidase to set up a potential chemically efficient biocatalyst. Glycobiology. Oxford: Oxford University Press, 2014, vol. 24, No 12, p. 1301-1311. ISSN 0959-6658. Available from: https://dx.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 = {http://dx.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 and Michaela WIMMEROVÁ. Protein engineering study of beta-mannosidase to set up a potential chemically efficient biocatalyst. \textit{Glycobiology}. Oxford: Oxford University Press, 2014, vol.~24, No~12, p.~1301-1311. ISSN~0959-6658. Available from: https://dx.doi.org/10.1093/glycob/cwu074.

Detailed Information on Publication Record