The invention discloses a recombinant
Pasteurella multocida-sourced
uridine diphosphate
glucuronide transferase (PmHS2)
mutant and a preparation method thereof, and relates to the technical field of
biology, the
mutant is obtained by
cloning a coding
gene to a pET26b (+) vector, transforming BL21 (DE3)
Escherichia coli competent cells, and then transforming the recombinant
Pasteurella multocida-sourced
uridine diphosphate
glucuronide transferase (PmHS2) to a pET26b (+) vector. And carrying out IPTG (
isopropyl-beta-d-thiogalactoside) induced culture, ultrasonication and
nickel affinity chromatography purification to obtain high-purity
protein. Compared with
wild type recombinant PmHS2, the activity of N-acetyl glucosamino
transferase of the
mutant is improved by 70%, the
residual activity is 67.1% after the mutant is stored at
room temperature for 5 days, the soluble expression quantity is improved by 6 times, and the mutant and the preparation method have the advantages of high
catalytic efficiency, high stability and high
solubility, and the mutant and the preparation method are mature in process, controllable in cost and suitable for industrial production. The method can be efficiently used in the fields of
heparin and derivative synthesis,
traditional Chinese medicine active ingredient glycosylation and the like, is suitable for large-scale production and has a wide application prospect.