Expression of arginine deiminase from Pseudomonas plecoglossicida CGMCC2039 in Escherichia coli and its anti-tumor activity.

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 degrees 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.

• Authors: Ye Ni, Zhenwei Li, Zhihao Sun, Pu Zheng, Yongmei Liu, Leilei Zhu, Ulrich Schwaneberg
• Journal: Current microbiology (Curr Microbiol) Vol. 58 Issue 6 Pg. 593-8 (Jun 2009) ISSN: 1432-0991 [Electronic] United States
• PMID: 19280262 (Publication Type: Journal Article)

Chemical References

• Antineoplastic Agents
• Bacterial Proteins
• Hydrolases
• arginine deiminase

Topics

• Amino Acid Sequence
• Animals
• Antineoplastic Agents (chemistry, metabolism, pharmacology)
• Bacterial Proteins (chemistry, genetics, metabolism, pharmacology)
• Cell Line, Tumor
• Cloning, Molecular
• Escherichia coli (genetics, metabolism)
• Female
• Gene Expression
• Humans
• Hydrolases (chemistry, genetics, metabolism, pharmacology)
• Mice
• Molecular Sequence Data
• Molecular Weight
• Neoplasms (drug therapy)
• Pseudomonas (chemistry, enzymology, genetics)
• Random Allocation
• Sequence Alignment
• Xenograft Model Antitumor Assays