Expression of Arginine Deiminase From Pseudomonas Plecoglossicida CGMCC2039 in Escherichia Coli and Its Anti-Tumor Activity

Curr Microbiol. 2009 Jun;58(6):593-8. doi: 10.1007/s00284-009-9376-0. Epub 2009 Mar 12.

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.

MeSH terms

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

Substances

  • Antineoplastic Agents
  • Bacterial Proteins
  • Hydrolases
  • arginine deiminase