Characterization of a New Marine Leucine Dehydrogenase from <i>Pseudomonas balearica</i> and Its Application for L-<i>tert</i>-Leucine Production
Leucine dehydrogenase (LeuDH) has emerged as the most promising biocatalyst for L-<i>tert</i>-leucine (L-Tle) production via asymmetric reduction in trimethylpyruvate (TMP). In this study, a new LeuDH named <i>Pb</i>LeuDH from marine <i>Pseudomonas balearica</i> w...
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MDPI AG
2022-08-01
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| author | Zewang Guo Denghui Chen Qi Xiong Miao Liang Pengfei Li Zehui Gong Junzhi Qiu Liaoyuan Zhang |
| author_facet | Zewang Guo Denghui Chen Qi Xiong Miao Liang Pengfei Li Zehui Gong Junzhi Qiu Liaoyuan Zhang |
| author_sort | Zewang Guo |
| collection | DOAJ |
| description | Leucine dehydrogenase (LeuDH) has emerged as the most promising biocatalyst for L-<i>tert</i>-leucine (L-Tle) production via asymmetric reduction in trimethylpyruvate (TMP). In this study, a new LeuDH named <i>Pb</i>LeuDH from marine <i>Pseudomonas balearica</i> was heterologously over-expressed in <i>Escherichia coli</i>, followed by purification and characterization. <i>Pb</i>LeuDH possessed a broad substrate scope, displaying activities toward numerous L-amino acids and <i>α</i>-keto acids. Notably, compared with those reported LeuDHs, <i>Pb</i>LeuDH exhibited excellent catalytic efficiency for TMP with a <i>K</i><sub>m</sub> value of 4.92 mM and a <i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub> value of 24.49 s<sup>−1</sup> mM<sup>−1</sup>. Subsequently, L-Tle efficient production was implemented from TMP by whole-cell biocatalysis using recombinant <i>E. coli</i> as a catalyst, which co-expressed <i>Pb</i>LeuDH and glucose dehydrogenase (GDH). Ultimately, using a fed-batch feeding strategy, 273 mM (35.8 g L<sup>−1</sup>) L-Tle was achieved with a 96.1% yield and 2.39 g L<sup>−1</sup> h<sup>−1</sup> productivity. In summary, our research provides a competitive biocatalyst for L-Tle green biosynthesis and lays a solid foundation for the realization of large-scale L-Tle industrial production. |
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| language | English |
| last_indexed | 2024-03-10T00:28:49Z |
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| series | Catalysts |
| spelling | doaj.art-a025585997d541ada11fa7ab82a4bf042023-11-23T15:30:10ZengMDPI AGCatalysts2073-43442022-08-0112997110.3390/catal12090971Characterization of a New Marine Leucine Dehydrogenase from <i>Pseudomonas balearica</i> and Its Application for L-<i>tert</i>-Leucine ProductionZewang Guo0Denghui Chen1Qi Xiong2Miao Liang3Pengfei Li4Zehui Gong5Junzhi Qiu6Liaoyuan Zhang7Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaKey Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaKey Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaKey Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaKey Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaKey Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaKey Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaKey Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaLeucine dehydrogenase (LeuDH) has emerged as the most promising biocatalyst for L-<i>tert</i>-leucine (L-Tle) production via asymmetric reduction in trimethylpyruvate (TMP). In this study, a new LeuDH named <i>Pb</i>LeuDH from marine <i>Pseudomonas balearica</i> was heterologously over-expressed in <i>Escherichia coli</i>, followed by purification and characterization. <i>Pb</i>LeuDH possessed a broad substrate scope, displaying activities toward numerous L-amino acids and <i>α</i>-keto acids. Notably, compared with those reported LeuDHs, <i>Pb</i>LeuDH exhibited excellent catalytic efficiency for TMP with a <i>K</i><sub>m</sub> value of 4.92 mM and a <i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub> value of 24.49 s<sup>−1</sup> mM<sup>−1</sup>. Subsequently, L-Tle efficient production was implemented from TMP by whole-cell biocatalysis using recombinant <i>E. coli</i> as a catalyst, which co-expressed <i>Pb</i>LeuDH and glucose dehydrogenase (GDH). Ultimately, using a fed-batch feeding strategy, 273 mM (35.8 g L<sup>−1</sup>) L-Tle was achieved with a 96.1% yield and 2.39 g L<sup>−1</sup> h<sup>−1</sup> productivity. In summary, our research provides a competitive biocatalyst for L-Tle green biosynthesis and lays a solid foundation for the realization of large-scale L-Tle industrial production.https://www.mdpi.com/2073-4344/12/9/971leucine dehydrogenaseL-<i>tert</i>-leucinetrimethylpyruvatereductive aminationwhole-cell biocatalysis |
| spellingShingle | Zewang Guo Denghui Chen Qi Xiong Miao Liang Pengfei Li Zehui Gong Junzhi Qiu Liaoyuan Zhang Characterization of a New Marine Leucine Dehydrogenase from <i>Pseudomonas balearica</i> and Its Application for L-<i>tert</i>-Leucine Production Catalysts leucine dehydrogenase L-<i>tert</i>-leucine trimethylpyruvate reductive amination whole-cell biocatalysis |
| title | Characterization of a New Marine Leucine Dehydrogenase from <i>Pseudomonas balearica</i> and Its Application for L-<i>tert</i>-Leucine Production |
| title_full | Characterization of a New Marine Leucine Dehydrogenase from <i>Pseudomonas balearica</i> and Its Application for L-<i>tert</i>-Leucine Production |
| title_fullStr | Characterization of a New Marine Leucine Dehydrogenase from <i>Pseudomonas balearica</i> and Its Application for L-<i>tert</i>-Leucine Production |
| title_full_unstemmed | Characterization of a New Marine Leucine Dehydrogenase from <i>Pseudomonas balearica</i> and Its Application for L-<i>tert</i>-Leucine Production |
| title_short | Characterization of a New Marine Leucine Dehydrogenase from <i>Pseudomonas balearica</i> and Its Application for L-<i>tert</i>-Leucine Production |
| title_sort | characterization of a new marine leucine dehydrogenase from i pseudomonas balearica i and its application for l i tert i leucine production |
| topic | leucine dehydrogenase L-<i>tert</i>-leucine trimethylpyruvate reductive amination whole-cell biocatalysis |
| url | https://www.mdpi.com/2073-4344/12/9/971 |
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