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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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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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
- Sulfobacillus acidophilus arginase
- l-ornithine production
- Neutral pH optimum
- Coupled-enzyme system