All rights reserved. The synthesized gene of ribokinase from Thermus species 2.9 was cloned in the expression vector pET-23b+. The effective overproduction strain expressing this enzyme in soluble form was created. The ribokinase purification method was developed. The specific activity of this enzyme were determined and the influence of different factors on the activity was examined. The kinetic parameters for basic natural substrates were calculated and also the substrate specificity for different carbohydrates of D-series was examined.