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Characterization of Two Novel Bacterial Type A exo-chitobiose Hydrolases Having C-terminal 5/12-type Carbohydrate-binding Modules

Type A chitinases (EC 3.2.1.14), GH family 18, attack chitin ((1 → 4)-2-acetamido-2-deoxy-β-d-glucan) and chito-oligosaccharides from the reducing end to catalyze release of chitobiose (N,N′-diacetylchitobiose) via hydrolytic cleavage of N-acetyl-β-d-glucosaminide (1 → 4)-β-linkages and are thus “ex...

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Main Authors: Shariza, Jamek, Nyffenegger, Christian, Muschiol, Jan, Holck, Jesper, Meyer, Anne S., Mikkelsen, Jørn D.
Format: Article
Language:English
English
Published: Springer Berlin Heidelberg 2017
Subjects:
TP Chemical technology
Online Access:http://umpir.ump.edu.my/id/eprint/17568/1/Characterization%20of%20two%20novel%20bacterial%20type%20A%20exo-chitobiose%20hydrolases%20having%20C-terminal%205-12-type%20carbohydrate-binding%20modules.pdf
http://umpir.ump.edu.my/id/eprint/17568/6/Characterization%20of%20two%20novel%20bacterial%20type%20A%20exo-chitobiose%20hydrolases%20having%20C-terminal%205-12-type%20carbohydrate-binding%20modules%201.pdf
http://umpir.ump.edu.my/id/eprint/17568/
https://doi.org/10.1007/s00253-017-8198-4
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spelling my.ump.umpir.175682018-05-15T07:18:19Z http://umpir.ump.edu.my/id/eprint/17568/ Characterization of Two Novel Bacterial Type A exo-chitobiose Hydrolases Having C-terminal 5/12-type Carbohydrate-binding Modules Shariza, Jamek Nyffenegger, Christian Muschiol, Jan Holck, Jesper Meyer, Anne S. Mikkelsen, Jørn D. TP Chemical technology Type A chitinases (EC 3.2.1.14), GH family 18, attack chitin ((1 → 4)-2-acetamido-2-deoxy-β-d-glucan) and chito-oligosaccharides from the reducing end to catalyze release of chitobiose (N,N′-diacetylchitobiose) via hydrolytic cleavage of N-acetyl-β-d-glucosaminide (1 → 4)-β-linkages and are thus “exo-chitobiose hydrolases.” In this study, the chitinase type A from Serratia marcescens (SmaChiA) was used as a template for identifying two novel exo-chitobiose hydrolase type A enzymes, FbalChi18A and MvarChi18A, originating from the marine organisms Ferrimonas balearica and Microbulbifer variabilis, respectively. Both FbalChi18A and MvarChi18A were recombinantly expressed in Escherichia coli and were confirmed to exert exo-chitobiose hydrolase activity on chito-oligosaccharides, but differed in temperature and pH activity response profiles. Amino acid sequence comparison of the catalytic β/α barrel domain of each of the new enzymes showed individual differences, but ~69% identity of each to that of SmaChiA and highly conserved active site residues. Superposition of a model substrate on 3D structural models of the catalytic domain of the enzymes corroborated exo-chitobiose hydrolase type A activity for FbalChi18A and MvarChi18A, i.e., substrate attack from the reducing end. A main feature of both of the new enzymes was the presence of C-terminal 5/12 type carbohydrate-binding modules (SmaChiA has no C-terminal carbohydrate binding module). These new enzymes may be useful tools for utilization of chitin as an N-acetylglucosamine donor substrate via chitobiose. Springer Berlin Heidelberg 2017-06 Article PeerReviewed application/pdf en http://umpir.ump.edu.my/id/eprint/17568/1/Characterization%20of%20two%20novel%20bacterial%20type%20A%20exo-chitobiose%20hydrolases%20having%20C-terminal%205-12-type%20carbohydrate-binding%20modules.pdf application/pdf en http://umpir.ump.edu.my/id/eprint/17568/6/Characterization%20of%20two%20novel%20bacterial%20type%20A%20exo-chitobiose%20hydrolases%20having%20C-terminal%205-12-type%20carbohydrate-binding%20modules%201.pdf Shariza, Jamek and Nyffenegger, Christian and Muschiol, Jan and Holck, Jesper and Meyer, Anne S. and Mikkelsen, Jørn D. (2017) Characterization of Two Novel Bacterial Type A exo-chitobiose Hydrolases Having C-terminal 5/12-type Carbohydrate-binding Modules. Applied Microbiology and Biotechnology, 101 (11). pp. 4533-4546. ISSN 0175-7598(print); 1432-0614(online) https://doi.org/10.1007/s00253-017-8198-4 DOI: 10.1007/s00253-017-8198-4
institution Universiti Malaysia Pahang
building UMP Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Pahang
content_source UMP Institutional Repository
url_provider http://umpir.ump.edu.my/
language English
English
topic TP Chemical technology
spellingShingle TP Chemical technology
Shariza, Jamek
Nyffenegger, Christian
Muschiol, Jan
Holck, Jesper
Meyer, Anne S.
Mikkelsen, Jørn D.
Characterization of Two Novel Bacterial Type A exo-chitobiose Hydrolases Having C-terminal 5/12-type Carbohydrate-binding Modules
description Type A chitinases (EC 3.2.1.14), GH family 18, attack chitin ((1 → 4)-2-acetamido-2-deoxy-β-d-glucan) and chito-oligosaccharides from the reducing end to catalyze release of chitobiose (N,N′-diacetylchitobiose) via hydrolytic cleavage of N-acetyl-β-d-glucosaminide (1 → 4)-β-linkages and are thus “exo-chitobiose hydrolases.” In this study, the chitinase type A from Serratia marcescens (SmaChiA) was used as a template for identifying two novel exo-chitobiose hydrolase type A enzymes, FbalChi18A and MvarChi18A, originating from the marine organisms Ferrimonas balearica and Microbulbifer variabilis, respectively. Both FbalChi18A and MvarChi18A were recombinantly expressed in Escherichia coli and were confirmed to exert exo-chitobiose hydrolase activity on chito-oligosaccharides, but differed in temperature and pH activity response profiles. Amino acid sequence comparison of the catalytic β/α barrel domain of each of the new enzymes showed individual differences, but ~69% identity of each to that of SmaChiA and highly conserved active site residues. Superposition of a model substrate on 3D structural models of the catalytic domain of the enzymes corroborated exo-chitobiose hydrolase type A activity for FbalChi18A and MvarChi18A, i.e., substrate attack from the reducing end. A main feature of both of the new enzymes was the presence of C-terminal 5/12 type carbohydrate-binding modules (SmaChiA has no C-terminal carbohydrate binding module). These new enzymes may be useful tools for utilization of chitin as an N-acetylglucosamine donor substrate via chitobiose.
format Article
author Shariza, Jamek
Nyffenegger, Christian
Muschiol, Jan
Holck, Jesper
Meyer, Anne S.
Mikkelsen, Jørn D.
author_facet Shariza, Jamek
Nyffenegger, Christian
Muschiol, Jan
Holck, Jesper
Meyer, Anne S.
Mikkelsen, Jørn D.
author_sort Shariza, Jamek
title Characterization of Two Novel Bacterial Type A exo-chitobiose Hydrolases Having C-terminal 5/12-type Carbohydrate-binding Modules
title_short Characterization of Two Novel Bacterial Type A exo-chitobiose Hydrolases Having C-terminal 5/12-type Carbohydrate-binding Modules
title_full Characterization of Two Novel Bacterial Type A exo-chitobiose Hydrolases Having C-terminal 5/12-type Carbohydrate-binding Modules
title_fullStr Characterization of Two Novel Bacterial Type A exo-chitobiose Hydrolases Having C-terminal 5/12-type Carbohydrate-binding Modules
title_full_unstemmed Characterization of Two Novel Bacterial Type A exo-chitobiose Hydrolases Having C-terminal 5/12-type Carbohydrate-binding Modules
title_sort characterization of two novel bacterial type a exo-chitobiose hydrolases having c-terminal 5/12-type carbohydrate-binding modules
publisher Springer Berlin Heidelberg
publishDate 2017
url http://umpir.ump.edu.my/id/eprint/17568/1/Characterization%20of%20two%20novel%20bacterial%20type%20A%20exo-chitobiose%20hydrolases%20having%20C-terminal%205-12-type%20carbohydrate-binding%20modules.pdf
http://umpir.ump.edu.my/id/eprint/17568/6/Characterization%20of%20two%20novel%20bacterial%20type%20A%20exo-chitobiose%20hydrolases%20having%20C-terminal%205-12-type%20carbohydrate-binding%20modules%201.pdf
http://umpir.ump.edu.my/id/eprint/17568/
https://doi.org/10.1007/s00253-017-8198-4
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score 13.211869

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