Abstract
This study aims to use neutral pH optimum arginase as the catalyst for high-efficiency l-ornithine production. Sulfobacillus acidophilus arginase was firstly cloned and overexpressed in Escherichia coli. The purified enzyme was obtained, and the molecular mass determination showed that this arginase was a hexamer. S. acidophilus arginase possessed similarities with the other arginases such as the conserved sequences, purification behavior, and the necessity for Mn2+ as a cofactor. The maximum enzyme activity was obtained at pH 7.5 and 70 °C. Thermostability and pH stability analysis showed that the arginase was stable at 30–60 °C and pH 7.0–8.5, respectively. The kinetic parameters suggested that S. acidophilus arginase could efficiently hydrolyze l-arginine. Bioconversion with this neutral pH optimum arginase had the advantages of avoiding producing by-product, high molar yield, and high-level production of l-ornithine. When the bioconversion was performed with a fed-batch strategy and a coupled-enzyme system involving S. acidophilus arginase and Jack bean urease, the final production of 2.87 mol/L was obtained with only 1.72 mmol/L l-arginine residue, and the molar yield was 99.9%. The highest production record suggests that S. acidophilus arginase has a great prospect in industrial l-ornithine production.
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (31701515) and Youth Scientific Talent Program of the National Grain Industry (LQ2018204).
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KH and XG conceived and designed research. KH and SRZ conducted experiments. SL contributed new reagents or analytical tools. KH, SRZ, HDS, and JL analyzed data. KH wrote the manuscript. All authors read and approved the manuscript.
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Highlights
1. Arginase from S. acidophilus was cloned and overexpressed in E. coli
2. This thermostable arginase had a neutral pH optimum
3. A urease-coupled system and the fed-batch strategy was used for l-ornithine production
4. This arginase and the conversion process have important industrial application potential
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Huang, K., Zhang, S., Guan, X. et al. Thermostable arginase from Sulfobacillus acidophilus with neutral pH optimum applied for high-efficiency l-ornithine production. Appl Microbiol Biotechnol 104, 6635–6646 (2020). https://doi.org/10.1007/s00253-020-10721-w
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DOI: https://doi.org/10.1007/s00253-020-10721-w