4 results about "Recombinant escherichia coli" patented technology

The invention discloses a purification and renaturation method for matrix metalloproteinase 13. The purification and renaturation method comprises the steps of performing fermentation of recombinant escherichia coli and inducible expression of a target protein; then performing bacteria crushing, inclusion body washing, and dissolving to obtain an extracting liquid containing the matrix metalloproteinase 13; performing purification on the extracting liquid by Ni2+/- affinity chromatography; and next, performing renaturation by using DEAE anion exchange chromatography or Desalting dimension exclusion chromatography. The purification and renaturation method is simple in operation and short in time consumption; each one liter of fermenting liquid can obtain more active proteins, so that the production cost is lowered; and the selling price of MMP-13 is reduced from current 120 RMB/[mu]g to 8-10 RMB/[mu]g.

The invention relates to a preparation method of fusion protein expressed by gene recombinant escherichia coli, which belongs to the technical field of bioengineering. Aiming at the defects in the preparation process of the fusion protein expressed by various escherichia coli engineering bacteria at present, particularly the difficulty of thallus cell breaking, a solution suitable for industrial mass production is developed. The method is mainly characterized in that after fermentation is finished, on the premise that thalli do not need to be collected in a centrifugal mode, the efficient thalli cell breaking effect is achieved in the mode that fermentation liquor is directly subjected to high-temperature thermal cell breaking, and then a supernate containing the recombinant fusion proteinis obtained through centrifugation of a disc centrifuge, a concentrated recombinant fusion protein solution is obtained by utilizing conventional processes of membrane filtration and membrane concentration, and then enzyme digestion is carried out to obtain the target polypeptide. The method is low in equipment requirement, simple in process flow, and low in cost, and very suitable for industriallarge-scale production.