BcoAI, a new site-specific endonuclease from Bacillus coagulans

N. N. Sokolov, M. A. Eldarov, N. V. Anikeitcheva, I. V. Karpychev, O. T. Samko, A. B. Fitzner, A. A. Kalugin, E. B. Choroshoutina, K. G. Skryabin

Institute of Biological and Medical Chemistry, Academy of Medical Sciences of the USSR, Moscow; Centre “Bioengineering”, V. A. Engelhardt Institute of Molecular Biology, Academy of Sciences of the USSR, Moscow

Abstract: A new site-specific endonuclease was detected in toluene lysates of Bacillus coagulans AUCM B-732 and designated as BcoAI. The enzyme was purified by fractionation of the cell-free extract in the two-phase PEG/dextran system followed by chromatography on DEAE-sepharose and phosphocellulose and shown to be free of nonspecific nucleases and phosphatases. BcoAI has three cleavage sites on λ DNA, but does not cleave SV40, pBR322 and pUC19 DNA. BcoAI recognizes the sequence 5' GAG ↓ GTG 3' on double-stranded DNA and cleaves it as indicated by the arrow to yield blunt-ended DNA fragments. Thus, BcoAI is a true isoschizomer of PmaCl from Pseudomonas maltophila C.

Russian Journal of Bioorganic Chemistry 1991, 17 (9):1188-1192

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