Site-specific endonuclease BbvBI from Bacillus brevis

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

Institute of Biological and Medical Chemistry
Russian Academy of Medical Sciences Russian, Moscow
*Centre «Bioengineering», V. A. Engelhardt Institute of Molecular Biology,
Russian Academy of Sciences, Moscow

Abstract: A new restriction endonuclease BbvBI free from contaminating nonspecific nucleases and phosphatases was isolated from the Bacillus brevis cells. The enzyme was purified by fractionating the sonicated cell-free extract in a two-phase PEG/dextran system and subsequent chromatographies on DEAE-sepharose, blue sepharose and heparin sepharose. The endonuclease BbvBI displayed the maximal activity at 45°C, pH between 8.0 and 8.5, MgCl₂ concentration in the range of 5–10 mM and at the low ionic strength. It is shown that the enzyme cleaves the sequence G'GYPC'C, with the preferential cleavage of GGTACC and GGCACC sites as compared with GGTGCC and GGCGCC. Thus, the restriction endonuclease BbvBI is a true isoschizomer of nuclease BanI.

Russian Journal of Bioorganic Chemistry 1992, 18 (1):47-51

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