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| Content Provider | Springer Nature Link |
|---|---|
| Author | Li, Hongbo Zhang, Lanwei Cui, Yanhua Luo, Xue Xue, Chaohui Wang, Shumei |
| Copyright Year | 2013 |
| Abstract | Transglutaminases (TGases) catalyze post-translational protein modifications by ε-(γ-glutamyl) links and covalent amide bonds. In plant, this enzyme is poorly studied and only the Zea mays TGase gene (tgz) has been cloned. The tgz had been expressed in Escherichia coli, but the recombinant protein was mainly present in inclusion bodies. Therefore, to obtain active, soluble protein, we optimized its coding sequence according to the codon bias of Pichia pastoris and synthesized the sequence with SOEing-PCR. The optimized fragment was successfully transformed into P. pastoris GS115 by electroporation. The optimal conditions for expression were under a final concentration of 0.5 % methanol and a time-course of 96 h. The synthesized recombinant Zea mays transglutaminase (TGZs) was purified by affinity method, its production was 4.4 mg/L, and the specific activity was 0.889 U/mg under optimal expression condition. Optimal activity for TGZs was observed at 37 °C and a pH of 8.0, respectively. The cross-linking reaction of TGZs to the casein was studied, and the result was same as the reaction of casein by microbial transglutaminase. These results indicated that an effective procedure for expressing and purifying TGZs in P. pastoris GS115 was established. |
| Starting Page | 939 |
| Ending Page | 947 |
| Page Count | 9 |
| File Format | |
| ISSN | 09593993 |
| Journal | World Journal of Microbiology and Biotechnology |
| Volume Number | 29 |
| Issue Number | 5 |
| e-ISSN | 15730972 |
| Language | English |
| Publisher | Springer Netherlands |
| Publisher Date | 2013-01-08 |
| Publisher Place | Dordrecht |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Transglutaminase Zea mays Codon optimization Pichia pastoris GS115 Applied Microbiology Biotechnology Biochemistry Environmental Engineering/Biotechnology Microbiology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Physiology Medicine Applied Microbiology and Biotechnology Biotechnology |
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