BmoI Restriction Endonuclease from the Bacterial Strain Brevundimonas mongoliensis 53 Recognizes the DNA Sequence 5’-GG^GWCCC-3’

D.A. Gonchar¹, M.A. Abdurashitov¹, V.A. Chernukhin¹, V.S. Dedkov¹, N.A. Mikhnenkova¹, V.E. Ryzhova¹, S.Kh. Degtyaryov¹

¹ SibEnzyme Ltd, Novosibirsk, Russia

* corresponding author: D.A. Gonchar, SibEnzyme Ltd, 2/12 Timakova Str.,
Novosibirsk 630117, Russia;
phone: +7(383)333-4991; fax: +7(383)333-6853;
E-mail: gonchar@sibenzyme.ru

We identified a bacterial strain, Brevundimonas mongoliensis 53, which produces a new Type II restriction endonuclease, designated BmoI.
BmoI recognizes the palindromic heptanucleotide sequence 5’-GG^GWCCC-3’ (W = A or T) and cleaves at the indicated position to generate three-nucleotide 5′ overhangs.
BmoI is an isoschizomer of the previously described restriction enzyme SanDI.
The producing strain was identified based on morphological and biochemical characteristics and by sequencing a fragment of the 16S rRNA gene.
A preparation of BmoI with an activity of 2000 U/ml was obtained using a three-step chromatographic purification workflow.
Optimal reaction conditions for BmoI were determined to be SE buffer W (10 mM Tris-HCl, pH 8.5; 10 mM MgCl₂; 100 mM NaCl; 1 mM DTT) and an incubation temperature of 37°C.

List of abbreviations:
BSA – bovine serum albumin;
DTT – dithiothreitol;
T7 DNA – bacteriophage T7 DNA;
λ DNA – bacteriophage λ DNA;
U – enzyme units;
Tris – tris(hydroxymethyl)aminomethane;
PAGE – polyacrylamide gel;
bp – base pairs;
RE – restriction endonuclease;
EDTA – ethylenediaminetetraacetic acid.

Keywords: producer strain, enzyme purification, restriction endonuclease, isoschizomer

DOI: 10.26213/3034-4301.2025.5.2.002

Citation:
Gonchar D.A., Abdurashitov M.A., Chernukhin V.A., Dedkov V.S., Mikhnenkova N.A., Ryzhova V.E., Degtyaryov S.Kh. (2025) BmoI restriction endonuclease from the bacterium Brevundimonas mongoliensis 53 recognizes the DNA sequence 5’-GG^GWCCC-3’. // DNA-Processing Enzymes, 2025(2), DOI: 10.26213/3034-4301.2025.5.2.002

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