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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| Format: | Article |
| Language: | English English |
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Springer Berlin Heidelberg
2017
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| Online Access: | http://umpir.ump.edu.my/id/eprint/17568/ 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 |
| _version_ | 1848820461647953920 |
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| 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 |
| building | UMP Institutional Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-15T02:09:49Z |
| format | Article |
| id | ump-17568 |
| institution | Universiti Malaysia Pahang |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T02:09:49Z |
| publishDate | 2017 |
| publisher | Springer Berlin Heidelberg |
| recordtype | eprints |
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| spelling | ump-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). (Published) https://doi.org/10.1007/s00253-017-8198-4 DOI: 10.1007/s00253-017-8198-4 |
| 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 |
| title | 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_short | 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 |
| topic | TP Chemical technology |
| url | http://umpir.ump.edu.my/id/eprint/17568/ http://umpir.ump.edu.my/id/eprint/17568/ http://umpir.ump.edu.my/id/eprint/17568/ 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 |