RU93018174A - METHOD FOR PRODUCING DEPOXYRIBONUCLEOTIDES - Google Patents

Claims (1)

The invention relates to microbiology and engineering enzymology, and in particular to methods for producing deoxyribonucleotides used in medicine and chemical and enzymatic synthesis of specific oligodeoxyribonucleotides used in genetic and molecular biological work using microorganisms and their enzymes. The method is carried out by growing without additional aeration of bacteria Propionibacterium freundenreichii ssp. shermanii and differs in that, 2 to 2.5 hours before the end of the cultivation, the antibiotic bleomycetin is introduced, then the cells are separated from the medium, resuspended in potassium phosphate buffer, permeabilized with X-100 triton, frozen in liquid nitrogen and quickly thawed, treated with glutaraldehyde and after dialysis is placed in an incubation medium containing a specific ribonucleotide. After 2 - 3 hours of incubation, the corresponding 2 ^{'-dezoksiribonukleozid} 5'-diphosphates isolated from the medium. The method allows one-step transformation resulting in directionally receive individual 2 ^{'-dezoksiribonukleozid-5'} -difosfaty (to specify the base) using a not previously used for such purposes in biotechnology cheaper substrates - ribonucleotides (yield relative to the substrate 25 - 37%).