Academic Journal
Construction and characterization of a novel glucose dehydrogenase-leucine dehydrogenase fusion enzyme for the biosynthesis of l-tert-leucine
| العنوان: | Construction and characterization of a novel glucose dehydrogenase-leucine dehydrogenase fusion enzyme for the biosynthesis of l-tert-leucine |
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| المؤلفون: | Liao, Langxing, Zhang, Yonghui, Wang, Yali, Fu, Yousi, Zhang, Aihui, Qiu, Ruodian, Yang, Shuhao, Fang, Baishan |
| المساهمون: | National Natural Science Foundation of China, Key Programme |
| المصدر: | Microbial Cell Factories ; volume 20, issue 1 ; ISSN 1475-2859 |
| بيانات النشر: | Springer Science and Business Media LLC |
| سنة النشر: | 2021 |
| الوصف: | Background Biosynthesis of l - tert -leucine ( l -tle), a significant pharmaceutical intermediate, by a cofactor regeneration system friendly and efficiently is a worthful goal all the time. The cofactor regeneration system of leucine dehydrogenase (LeuDH) and glucose dehydrogenase (GDH) has showed great coupling catalytic efficiency in the synthesis of l -tle, however the multi-enzyme complex of GDH and LeuDH has never been constructed successfully. Results In this work, a novel fusion enzyme (GDH–R3–LeuDH) for the efficient biosynthesis of l -tle was constructed by the fusion of LeuDH and GDH mediated with a rigid peptide linker. Compared with the free enzymes, both the environmental tolerance and thermal stability of GDH–R3–LeuDH had a great improved since the fusion structure. The fusion structure also accelerated the cofactor regeneration rate and maintained the enzyme activity, so the productivity and yield of l -tle by GDH–R3–LeuDH was all enhanced by twofold. Finally, the space–time yield of l -tle catalyzing by GDH–R3–LeuDH whole cells could achieve 2136 g/L/day in a 200 mL scale system under the optimal catalysis conditions (pH 9.0, 30 ° C, 0.4 mM of NAD + and 500 mM of a substrate including trimethylpyruvic acid and glucose). Conclusions It is the first report about the fusion of GDH and LeuDH as the multi-enzyme complex to synthesize l -tle and reach the highest space–time yield up to now. These results demonstrated the great potential of the GDH–R3–LeuDH fusion enzyme for the efficient biosynthesis of l -tle. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1186/s12934-020-01501-2 |
| DOI: | 10.1186/s12934-020-01501-2.pdf |
| DOI: | 10.1186/s12934-020-01501-2/fulltext.html |
| الاتاحة: | http://dx.doi.org/10.1186/s12934-020-01501-2 http://link.springer.com/content/pdf/10.1186/s12934-020-01501-2.pdf http://link.springer.com/article/10.1186/s12934-020-01501-2/fulltext.html |
| Rights: | http://creativecommons.org/licenses/by/4.0/ ; http://creativecommons.org/licenses/by/4.0/ |
| رقم الانضمام: | edsbas.134C8539 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1186/s12934-020-01501-2 |
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