PMID- 19887741
OWN - NLM
STAT- MEDLINE
DCOM- 20091217
LR  - 20190606
IS  - 0916-7250 (Print)
IS  - 0916-7250 (Linking)
VI  - 71
IP  - 10
DP  - 2009 Oct
TI  - Growth inhibition of human melanoma cells by a recombinant arginine deiminase 
      expressed in Escherichia coli.
PG  - 1343-7
AB  - We have cloned the arginine deiminase (ADI) gene from Mycoplasma hominis PG21 
      genomic DNA by polymerase chain reaction, and changed four TGA tryptophan codons 
      (stop codon in E. coli) to TGG codons in the coding region by site-directed 
      mutagenesis in order to express in E. coli. The recombinant ADI (rADI) was 
      purified to apparent homogeneity by Ni-affinity chromatography after extraction 
      from inclusion bodies followed by refolding. The rADI expressed in E. coli was 
      estimated to be 50 kDa. Dimeric forms of rADI exerted enzymatic activity. We 
      found that high concentration of potassium dihydrogenphosphate (PDP) and 
      L-arginine addition in refolding reaction increases the enzyme activity. The 
      specific activity of rADl was calculated as 0.618 U/mg. In addition, the enzyme 
      activity of purified rADI remained for at least one month in 100 mM PDP solution 
      (pH 6.5), but diminished within one week in 100 mM PDP solution (pH 7.4). 
      Anti-tumor activity of the purified rADI was estimated to be 0.036 U/ml as 50% 
      growth inhibitory activity against human melanoma cell line G-361.
FAU - Kozai, Megumi
AU  - Kozai M
AD  - Department of Veterinary Microbiology, Faculty of Agriculture, Iwate University, 
      3-18-8 Ueda, Morioka 020-8550, Japan.
FAU - Sasamori, Eriko
AU  - Sasamori E
FAU - Fujihara, Masatoshi
AU  - Fujihara M
FAU - Yamashita, Tetsuro
AU  - Yamashita T
FAU - Taira, Hideharu
AU  - Taira H
FAU - Harasawa, Ryo
AU  - Harasawa R
LA  - eng
PT  - Journal Article
PL  - Japan
TA  - J Vet Med Sci
JT  - The Journal of veterinary medical science
JID - 9105360
RN  - 0 (Antineoplastic Agents)
RN  - 0 (Recombinant Proteins)
RN  - EC 3.- (Hydrolases)
RN  - EC 3.5.3.6 (arginine deiminase)
SB  - IM
MH  - Antineoplastic Agents/pharmacology
MH  - Cell Line, Tumor
MH  - Cloning, Molecular
MH  - Escherichia coli/genetics/metabolism
MH  - Gene Expression Regulation, Bacterial
MH  - Gene Expression Regulation, Enzymologic
MH  - Humans
MH  - Hydrolases/chemistry/genetics/*metabolism/*pharmacology
MH  - Melanoma/*metabolism
MH  - Mycoplasma hominis/*enzymology
MH  - *Recombinant Proteins
EDAT- 2009/11/06 06:00
MHDA- 2009/12/18 06:00
CRDT- 2009/11/06 06:00
PHST- 2009/11/06 06:00 [entrez]
PHST- 2009/11/06 06:00 [pubmed]
PHST- 2009/12/18 06:00 [medline]
AID - JST.JSTAGE/jvms/001343 [pii]
AID - 10.1292/jvms.001343 [doi]
PST - ppublish
SO  - J Vet Med Sci. 2009 Oct;71(10):1343-7. doi: 10.1292/jvms.001343.