Construction Of PXJ128 Gene-Engineering Strain And Study On Characteristic Of Acetylornithine Deacetylase

This research is a part work of the "863" International project- "The Technique of Producing L-Methionine by Gene-Engineering Strain of Immobilization Cell" . Acetylornithine deacetylase is the key enzyme of producting L-methionine.we mainly do research work on the construction of Acetylornithine deacetylase gene-engineering strain and Characteristic of Proteinase.In order to get high expression Deacetylase strain,we obtain the gene by PCR ArgE gene.The product(2800bp) was cloned into PUC19 plasmid and confirmed with blue/white dot screening > restriction enzyme analysis and PCR.Then taking the nucleotide sequencing compared with the sequence at BLAST of U.S.A.We constructed a high expression of gene-engineering strain-PXJ 128 which containing the ArgE gene on the high expressing system of PXJI18 with activity of Acetylornithine deacetylase above 20000u/g. At mean time,we have studied on the characteristic of proteinase and make sure the suitable condition and influencing factor of deacetylase function. Using the technology of immobiolized cell,the utlization of deacetylase have been increased deeply and lower production cost.The half deline period is more than 20 days and the production of L-Methionine is above 75% ?Meanwhile, in order to improve the E.coli with ability of using sugar. We have recombined the vecter of PARG25 A which have ability of using sugar with the gene of Sucrase length of 7800bp .