Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli
The full length phytase gene of Mitsuokella jalaludinii was successfully cloned and was found to be 1047 bp in length, with 348 amino acids, and was designated as PHY7 phytase gene. A comparison of the sequence of PHY7 phytase gene of M. jalaludinii with various microbial phytase gene sequences show...
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| Format: | Article |
| Language: | English |
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Elsevier
2015
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| Online Access: | http://psasir.upm.edu.my/id/eprint/36797/ http://psasir.upm.edu.my/id/eprint/36797/1/Cloning%20of%20a%20novel%20phytase%20from%20an%20anaerobic%20rumen%20bacterium%2C%20Mitsuokella%20jalaludinii%2C%20and%20its%20expression%20in%20Escherichia%20coli.pdf |
| _version_ | 1848848432205135872 |
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| author | Tan, Wan Qin Phang, Chiun Yee Sieo, Chin Chin Yiap, Beow Chin Wong, Clemente Michael Vui Ling Abdullah, Norhani Radu, Son Ho, Yin Wan |
| author_facet | Tan, Wan Qin Phang, Chiun Yee Sieo, Chin Chin Yiap, Beow Chin Wong, Clemente Michael Vui Ling Abdullah, Norhani Radu, Son Ho, Yin Wan |
| author_sort | Tan, Wan Qin |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | The full length phytase gene of Mitsuokella jalaludinii was successfully cloned and was found to be 1047 bp in length, with 348 amino acids, and was designated as PHY7 phytase gene. A comparison of the sequence of PHY7 phytase gene of M. jalaludinii with various microbial phytase gene sequences showed that it was not similar to those from other bacteria except Selenomonas ruminatium, thus suggesting that they may both express a new class of phytase. The PHY7 phytase gene was subsequently subcloned into bacterial expression vector, pET32a, for expression in Escherichia coli strain Rosetta-gami. Expression of the recombinant phytase gene was optimized and characterized. The recombinant phytase was estimated to be approximately 55 kDa by SDS-PAGE analysis. The recombinant phytase exhibited optimum activity at 55°C, pH 4.5 and showed good pH stability from pH 3.5 to 5.5 (>78% relative activity). Metal ions such as Ca2+, Mg2+, and K+ were found to exert significant stimulatory effect on the recombinant phytase activity while Cu2+, Fe3+, and Zn2+ greatly inhibited the enzyme activity. The recombinant phytase showed moderate resistance to trypsin proteolysis, but susceptible to pepsin proteolysis. The results of the study showed that several characteristics of recombinant phytase were slightly different from the native enzyme. Unfavourable characteristics such as reduced pH stability and metal ion effects should be taken into consideration during feed enzyme formulation. |
| first_indexed | 2025-11-15T09:34:24Z |
| format | Article |
| id | upm-36797 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T09:34:24Z |
| publishDate | 2015 |
| publisher | Elsevier |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-367972015-12-15T03:39:35Z http://psasir.upm.edu.my/id/eprint/36797/ Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli Tan, Wan Qin Phang, Chiun Yee Sieo, Chin Chin Yiap, Beow Chin Wong, Clemente Michael Vui Ling Abdullah, Norhani Radu, Son Ho, Yin Wan The full length phytase gene of Mitsuokella jalaludinii was successfully cloned and was found to be 1047 bp in length, with 348 amino acids, and was designated as PHY7 phytase gene. A comparison of the sequence of PHY7 phytase gene of M. jalaludinii with various microbial phytase gene sequences showed that it was not similar to those from other bacteria except Selenomonas ruminatium, thus suggesting that they may both express a new class of phytase. The PHY7 phytase gene was subsequently subcloned into bacterial expression vector, pET32a, for expression in Escherichia coli strain Rosetta-gami. Expression of the recombinant phytase gene was optimized and characterized. The recombinant phytase was estimated to be approximately 55 kDa by SDS-PAGE analysis. The recombinant phytase exhibited optimum activity at 55°C, pH 4.5 and showed good pH stability from pH 3.5 to 5.5 (>78% relative activity). Metal ions such as Ca2+, Mg2+, and K+ were found to exert significant stimulatory effect on the recombinant phytase activity while Cu2+, Fe3+, and Zn2+ greatly inhibited the enzyme activity. The recombinant phytase showed moderate resistance to trypsin proteolysis, but susceptible to pepsin proteolysis. The results of the study showed that several characteristics of recombinant phytase were slightly different from the native enzyme. Unfavourable characteristics such as reduced pH stability and metal ion effects should be taken into consideration during feed enzyme formulation. Elsevier 2015-09 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/36797/1/Cloning%20of%20a%20novel%20phytase%20from%20an%20anaerobic%20rumen%20bacterium%2C%20Mitsuokella%20jalaludinii%2C%20and%20its%20expression%20in%20Escherichia%20coli.pdf Tan, Wan Qin and Phang, Chiun Yee and Sieo, Chin Chin and Yiap, Beow Chin and Wong, Clemente Michael Vui Ling and Abdullah, Norhani and Radu, Son and Ho, Yin Wan (2015) Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli. Journal of Integrative Agriculture, 14 (9). pp. 1816-1826. ISSN 2095-3119; ESSN: 2352-3425 http://www.sciencedirect.com/science/article/pii/S2095311914609606 10.1016/S2095-3119(14)60960-6 |
| spellingShingle | Tan, Wan Qin Phang, Chiun Yee Sieo, Chin Chin Yiap, Beow Chin Wong, Clemente Michael Vui Ling Abdullah, Norhani Radu, Son Ho, Yin Wan Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli |
| title | Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli |
| title_full | Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli |
| title_fullStr | Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli |
| title_full_unstemmed | Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli |
| title_short | Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli |
| title_sort | cloning of a novel phytase from an anaerobic rumen bacterium, mitsuokella jalaludinii, and its expression in escherichia coli |
| url | http://psasir.upm.edu.my/id/eprint/36797/ http://psasir.upm.edu.my/id/eprint/36797/ http://psasir.upm.edu.my/id/eprint/36797/ http://psasir.upm.edu.my/id/eprint/36797/1/Cloning%20of%20a%20novel%20phytase%20from%20an%20anaerobic%20rumen%20bacterium%2C%20Mitsuokella%20jalaludinii%2C%20and%20its%20expression%20in%20Escherichia%20coli.pdf |