Cloning, production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite Coptotermes curvignathus
Coptotermes curvignathus is a termite that, owing to its ability to digest living trees, serves as a gold mine for robust industrial enzymes. This unique characteristic reflects the presence of very efficient hydrolytic enzyme systems including cellulases. Transcriptomic analyses of the gut of C. cu...
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| Format: | Article |
| Language: | English |
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Humana Press
2017
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| Online Access: | http://psasir.upm.edu.my/id/eprint/61129/1/Cloning%2C%20production%20and%20characterization%20of%20a%20glycoside%20hydrolase%20family%207%20enzyme%20from%20the%20gut%20microbiota%20of%20the%20termite%20Coptotermes%20curvignathus.pdf |
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| author | Woon, James Sy-Keen King, Patricia Jie Hung Mackeen, Mukram Mohamed Mahadi, Nor Muhammad Wan Seman, Wan Mohd Khairulikhsan Broughton, William J. Abdul Murad, Abdul Munir Abu Bakar, Farah Diba |
| author_facet | Woon, James Sy-Keen King, Patricia Jie Hung Mackeen, Mukram Mohamed Mahadi, Nor Muhammad Wan Seman, Wan Mohd Khairulikhsan Broughton, William J. Abdul Murad, Abdul Munir Abu Bakar, Farah Diba |
| author_sort | Woon, James Sy-Keen |
| collection | UPM |
| description | Coptotermes curvignathus is a termite that, owing to its ability to digest living trees, serves as a gold mine for robust industrial enzymes. This unique characteristic reflects the presence of very efficient hydrolytic enzyme systems including cellulases. Transcriptomic analyses of the gut of C. curvignathus revealed that carbohydrate-active enzymes (CAZy) were encoded by 3254 transcripts and that included 69 transcripts encoding glycoside hydrolase family 7 (GHF7) enzymes. Since GHF7 enzymes are useful to the biomass conversion industry, a gene encoding for a GHF7 enzyme (Gh1254) was synthesized, sub-cloned and expressed in the methylotrophic yeast Pichia pastoris. Expressed GH1254 had an apparent molecular mass of 42 kDa, but purification was hampered by its low expression levels in shaken flasks. To obtain more of the enzyme, GH1254 was produced in a bioreactor that resulted in a fourfold increase in crude enzyme levels. The purified enzyme was active towards soluble synthetic substrates such as 4-methylumbelliferyl-β-d-cellobioside, 4-nitrophenyl-β-d-cellobioside and 4-nitrophenyl-β-d-lactoside but was non-hydrolytic towards Avicel or carboxymethyl cellulose. GH1254 catalyzed optimally at 35 °C and maintained 70% of its activity at 25 °C. This enzyme is thus potentially useful in food industries employing low-temperature conditions. |
| first_indexed | 2024-03-06T09:39:45Z |
| format | Article |
| id | upm.eprints-61129 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:39:45Z |
| publishDate | 2017 |
| publisher | Humana Press |
| record_format | dspace |
| spelling | upm.eprints-611292018-09-14T03:27:55Z http://psasir.upm.edu.my/id/eprint/61129/ Cloning, production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite Coptotermes curvignathus Woon, James Sy-Keen King, Patricia Jie Hung Mackeen, Mukram Mohamed Mahadi, Nor Muhammad Wan Seman, Wan Mohd Khairulikhsan Broughton, William J. Abdul Murad, Abdul Munir Abu Bakar, Farah Diba Coptotermes curvignathus is a termite that, owing to its ability to digest living trees, serves as a gold mine for robust industrial enzymes. This unique characteristic reflects the presence of very efficient hydrolytic enzyme systems including cellulases. Transcriptomic analyses of the gut of C. curvignathus revealed that carbohydrate-active enzymes (CAZy) were encoded by 3254 transcripts and that included 69 transcripts encoding glycoside hydrolase family 7 (GHF7) enzymes. Since GHF7 enzymes are useful to the biomass conversion industry, a gene encoding for a GHF7 enzyme (Gh1254) was synthesized, sub-cloned and expressed in the methylotrophic yeast Pichia pastoris. Expressed GH1254 had an apparent molecular mass of 42 kDa, but purification was hampered by its low expression levels in shaken flasks. To obtain more of the enzyme, GH1254 was produced in a bioreactor that resulted in a fourfold increase in crude enzyme levels. The purified enzyme was active towards soluble synthetic substrates such as 4-methylumbelliferyl-β-d-cellobioside, 4-nitrophenyl-β-d-cellobioside and 4-nitrophenyl-β-d-lactoside but was non-hydrolytic towards Avicel or carboxymethyl cellulose. GH1254 catalyzed optimally at 35 °C and maintained 70% of its activity at 25 °C. This enzyme is thus potentially useful in food industries employing low-temperature conditions. Humana Press 2017 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/61129/1/Cloning%2C%20production%20and%20characterization%20of%20a%20glycoside%20hydrolase%20family%207%20enzyme%20from%20the%20gut%20microbiota%20of%20the%20termite%20Coptotermes%20curvignathus.pdf Woon, James Sy-Keen and King, Patricia Jie Hung and Mackeen, Mukram Mohamed and Mahadi, Nor Muhammad and Wan Seman, Wan Mohd Khairulikhsan and Broughton, William J. and Abdul Murad, Abdul Munir and Abu Bakar, Farah Diba (2017) Cloning, production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite Coptotermes curvignathus. Molecular Biotechnology, 59 (7). 271 - 283. ISSN 1073-6085; ESSN: 1559-0305 https://link.springer.com/content/pdf/10.1007%2Fs12033-017-0015-x.pdf 10.1007/s12033-017-0015-x |
| spellingShingle | Woon, James Sy-Keen King, Patricia Jie Hung Mackeen, Mukram Mohamed Mahadi, Nor Muhammad Wan Seman, Wan Mohd Khairulikhsan Broughton, William J. Abdul Murad, Abdul Munir Abu Bakar, Farah Diba Cloning, production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite Coptotermes curvignathus |
| title | Cloning, production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite Coptotermes curvignathus |
| title_full | Cloning, production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite Coptotermes curvignathus |
| title_fullStr | Cloning, production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite Coptotermes curvignathus |
| title_full_unstemmed | Cloning, production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite Coptotermes curvignathus |
| title_short | Cloning, production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite Coptotermes curvignathus |
| title_sort | cloning production and characterization of a glycoside hydrolase family 7 enzyme from the gut microbiota of the termite coptotermes curvignathus |
| url | http://psasir.upm.edu.my/id/eprint/61129/1/Cloning%2C%20production%20and%20characterization%20of%20a%20glycoside%20hydrolase%20family%207%20enzyme%20from%20the%20gut%20microbiota%20of%20the%20termite%20Coptotermes%20curvignathus.pdf |
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