Improved broad-spectrum endonuclease and industrial production method thereof

Abstract

The invention relates to DNA which achieves efficient expression in yeast cells and is used for coding improved broad-spectrum endonuclease, the improved broad-spectrum endonuclease which is coded by the DNA, and a method for constructing the DNA and the broad-spectrum endonuclease by virtue of genetic engineering technology and protein engineering technology; and the invention also relates to an industrial fermentation method for producing the improved broad-spectrum endonuclease. By virtue of the industrial fermentation method, the DNA, which is used for coding the improved broad-spectrum endonuclease, is expressed in eukaryotic host yeast cells, and the produced improved broad-spectrum endonuclease is high in specific activity and yield; therefore, the problems of the prior art that yield is low and an operation of conducting purifying is difficult. In addition, the improved broad-spectrum endonuclease disclosed by the invention, which is free from bacterial endotoxin, is conducive to application of medical and bio-engineering fields. Meanwhile, the improved broad-spectrum endonuclease disclosed by the invention can be also prepared into a dosage form of freeze-dried powder, so that the broad-spectrum endonuclease, as a finished product, is more convenient for transportation, preservation and industrial application.

Description

technical field [0001] The present invention relates to a DNA encoding a broad-spectrum endonuclease, a broad-spectrum endonuclease encoded by the DNA, and a method for constructing the DNA and a broad-spectrum endonuclease by using genetic engineering and protein engineering techniques ; The present invention also relates to the industrial fermentation method of producing this broad-spectrum endonuclease. Specifically, the present invention relates to a DNA encoding an improved Prodigo nuclease, an improved Prodigo nuclease encoded by the DNA, and a method for constructing the DNA and Prodigo nuclease using genetic engineering and protein engineering techniques ; The present invention also relates to an industrial fermentation process for producing the improved Prodigo nuclease. Background technique [0002] As a broad-spectrum endonuclease, Prodigo nuclease is a non-restricted broad-spectrum endonuclease derived from Serratia marcescens (also known as Prodigy), which can ...

AI Technical Summary

Problems solved by technology

However, because Benzonase is toxic to E. coli host cells and the purification method is complicated, the yield is low and the price is high

The price of foreign products is as high as 30,000 to 80,000 yuan (Sigma and Merck) per 1 million units (equivalent to about 1 mg of protein), and the price of similar domestic products is also higher than 10,000 yuan. Such high prices limit the Wide application of Prodigobacterium nuclease

In addition, because it is not easy to remove the bacterial endotoxin of the prokaryotic expression system from the prodigy nuclease product, the application of prodigo nuclease in industrial production is limited.

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