دورية أكاديمية
The identification of a robust leucine dehydrogenase from a directed soil metagenome for efficient synthesis of L-2-aminobutyric acid.
العنوان: | The identification of a robust leucine dehydrogenase from a directed soil metagenome for efficient synthesis of L-2-aminobutyric acid. |
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المؤلفون: | Liu Y; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China., Zhong X; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China., Luo Z; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China., Meng X; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China., Li R; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China., Zhong W; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China., Yang L; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China., Wang H; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China., Wei D; State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China. |
المصدر: | Biotechnology journal [Biotechnol J] 2023 Aug; Vol. 18 (8), pp. e2200590. Date of Electronic Publication: 2023 May 15. |
نوع المنشور: | Journal Article |
اللغة: | English |
بيانات الدورية: | Publisher: Wiley-VCH Verlag Country of Publication: Germany NLM ID: 101265833 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1860-7314 (Electronic) Linking ISSN: 18606768 NLM ISO Abbreviation: Biotechnol J Subsets: MEDLINE |
أسماء مطبوعة: | Original Publication: Weinheim : Wiley-VCH Verlag, c2006- |
مواضيع طبية MeSH: | Metagenome* , Aminobutyrates*/metabolism, Leucine Dehydrogenase/genetics ; Leucine Dehydrogenase/metabolism ; Biotechnology ; Leucine |
مستخلص: | L-2-aminobutyric acid (L-2-ABA) is a chiral precursor for the synthesis of anti-epileptic drug levetiracetam and anti-tuberculosis drug ethambutol. Asymmetric synthesis of L-2-ABA by leucine dehydrogenases has been widely developed. However, the limitations of natural enzymes, such as poor stability, low catalytic efficiency, and inhibition of high-concentration substrates, limit large-scale applications. Herein, by directed screening of a metagenomic library from unnatural amino acid-enriched environments, a robust leucine dehydrogenase, TvLeuDH, was identified, which exhibited high substrate tolerance and excellent enzymatic activity towards 2-oxobutyric acid. In addition, TvLeuDH has strong affinity for NADH. Subsequently, a three-enzyme co-expression system containing L-threonine deaminase, TvLeuDH, and glucose dehydrogenase was established. By optimizing reaction conditions, 1.5 M L-threonine could be converted to L-2-ABA with a 99% molar conversion rate and a space-time yield of 51.5 g·L -1 ·h -1 . In this process, no external coenzyme was added. The robustness of TvLeuDH allowed the reaction to be performed without the addition of extra salt as the buffer, demonstrating the simplest reaction system currently reported. These unique properties for the efficient and environmentally friendly production of chiral amino acids make TvLeuDH a particularly promising candidate for industrial applications, which reveals the great potential of directed metagenomics for industrial biotechnology. (© 2023 Wiley-VCH GmbH.) |
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فهرسة مساهمة: | Keywords: L-2-aminobutyric acid; leucine dehydrogenase; metagenomics; substrate tolerance |
المشرفين على المادة: | EC 1.4.1.9 (Leucine Dehydrogenase) 8306QPJ19P (alpha-aminobutyric acid) 0 (Aminobutyrates) GMW67QNF9C (Leucine) |
تواريخ الأحداث: | Date Created: 20230507 Date Completed: 20230817 Latest Revision: 20230817 |
رمز التحديث: | 20230817 |
DOI: | 10.1002/biot.202200590 |
PMID: | 37149736 |
قاعدة البيانات: | MEDLINE |
تدمد: | 1860-7314 |
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DOI: | 10.1002/biot.202200590 |