Expression of Arginine Deiminase from Pseudomonas plecoglossicida CGMCC2039 in Escherichia coli and Its Anti-Tumor Activity |
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Authors: | Ye Ni Zhenwei Li Zhihao Sun Pu Zheng Yongmei Liu Leilei Zhu Ulrich Schwaneberg |
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Affiliation: | (1) School of Biotechnology, Jiangnan University, Wuxi, Jiansu, China;(2) School of Engineering and Science, Jacobs University Bremen, Bremen, Germany |
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Abstract: | Arginine deiminase (ADI), an arginine-degrading enzyme, has been studied as a potential anti-cancer agent in clinical trials for the treatment of arginine-auxotrophic tumors, such as hepatocellular carcinomas (HCCs) and melanomas. The arcA gene encoding ADI was cloned from a recently isolated strain Pseudomonas plecoglossicida CGMCC2039. The nucleotide sequence of ADI comprises an ORF of 1,254 bp encoding 417 amino acids. The deduced ADI protein sequence has a calculated molecular weight of 46.5 kDa and shows 97% and 85% identity to ADIs from P. putida and P. aeruginosa, respectively. The arcA from P. plecoglossicida CGMCC2039 was expressed in Escherichia coli BL21 with a N-terminal His6-tag, and purified to homogeneity. A molecular mass of approximate 49 kDa was confirmed by SDS-PAGE analysis and specific activity was determined to be 4.76 U/mg (pH 6.0 and 37°C). In vivo activity study showed that the rADI could effectively inhibit H22 tumor growth at a total dose of 5 U/mouse over a 2-week dosing period. |
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