Academic Journal
Protein engineering of NADH pyrophosphatase for efficient biocatalytic production of reduced nicotinamide mononucleotide
| العنوان: | Protein engineering of NADH pyrophosphatase for efficient biocatalytic production of reduced nicotinamide mononucleotide |
|---|---|
| المؤلفون: | Liu, Ye, Gong, Jin-Song, Marshall, George, Su, Chang, Hall, Michael, Li, Heng, Xu, Guo-Qiang, Shi, Jin-Song, Xu, Zheng-Hong |
| المصدر: | Frontiers in Bioengineering and Biotechnology ; volume 11 ; ISSN 2296-4185 |
| بيانات النشر: | Frontiers Media SA |
| سنة النشر: | 2023 |
| المجموعة: | Frontiers (Publisher - via CrossRef) |
| الوصف: | Introduction: NADH pyrophosphatase, a hydrolase catalyzing the phosphate bond of NADH to reduced nicotinamide mononucleotide, has potential applications in the food, cosmetic and pharmaceutical industry. Methods: Here, we investigated the effects of vector screening, promoter and RBS strategies on NADH pyrophosphatase expression and protein engineering on its enzymatic activity and thermal stability. Results: In this study, we describe a NADH pyrophosphatase derived from Escherichia coli ( EcNudc ). Strategies focusing on expression regulation including screening vectors, optimizing promoters and ribosome binding sites were utilized to enhance the productivity of EcNudc (1.8 U/mL). Moreover, protein engineering was adopted to further improve the catalytic properties of EcNudc , achieving 3.3-fold higher activity and 3.6-fold greater thermostability at 50°C. Furthermore, fermentation for the combined mutant R148A-H149E ( EcNudc-M ) production in a 7 L fermenter was implemented and the enzyme activity of EcNudc-M reached 33.0 U/mL. Finally, the EcNudc-M was applied in the catalysis of NADH with the highest NMNH yield of 16.65 g/L. Discussion: In conclusion, we constructed a commercially available genetically engineered strain with high activity and thermal stability of NADH pyrophosphatase, laying a broad foundation for the biocatalytic industrial production of NMNH and expand its application range. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | unknown |
| DOI: | 10.3389/fbioe.2023.1159965 |
| DOI: | 10.3389/fbioe.2023.1159965/full |
| الاتاحة: | http://dx.doi.org/10.3389/fbioe.2023.1159965 https://www.frontiersin.org/articles/10.3389/fbioe.2023.1159965/full |
| Rights: | https://creativecommons.org/licenses/by/4.0/ |
| رقم الانضمام: | edsbas.CB45673D |
| قاعدة البيانات: | BASE |
| DOI: | 10.3389/fbioe.2023.1159965 |
|---|