Restriction endonuclease Aor13HI from Acidiphilium organovorum 13H, a new isoschizomer of BspMII: purification and characterization.
A restriction endonuclease, Aor13HI, an isoschizomer of BspMII, was purified to homogeneity from cell extracts of Acidiphilium organovorum strain 13H. The enzyme has a molecular mass of 60,000 daltons and consists of two subunits identical in molecular mass of 30,000 daltons. Aor13HI endonuclease, like BspMII, recognizes the palindromic six-base sequence 5'-TCCGGA-3', and cleaves between the T and C to produce a four-base 5' extension. Aor13HI is not inhibited by dam-dependent methylation. The isoelectric point of the enzyme is 5.7. Aor13HI activity was maximum at pH 7.5, 100 mM KCl, 7.5-10 mM MgCl2, and 55 degrees C. The enzyme was stable up to 60 degrees C. The N-terminal amino acid sequence (30 residues) of Aor13HI did not show any similarity with the sequence of other restriction endonucleases reported.
Inagaki
K
K
Division of Bioresources Science, Faculty of Agriculture, Okayama University, Japan.
Hikita
T
T
Yanagidani
S
S
Nomura
Y
Y
Kishimoto
N
N
Tano
T
T
Tanaka
H
H
D000595:Amino Acid Sequence; D001665:Binding Sites; D002850:Chromatography, Gel; D002942:Circular Dichroism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006091:Gram-Negative Chemolithotrophic Bacteria / Q000201:enzymology* / Q000254:growth & development; D007526:Isoelectric Point; D008745:Methylation; D008969:Molecular Sequence Data; D008970:Molecular Weight
D015252:Deoxyribonucleases, Type II Site-Specific; C053919:TCCGGA-specific type II deoxyribonucleases