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streptomycin

kanamycin

→

L-lysine

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후처리

1. 1
   workup.ADDITIONasd, lysA, and ddh genes were introduced into the chromosome by the method

실험 절차

pAET7 was introduced into V12Sα obtained in Example 16, and the pMIV-FRTKmFRT-EABDAS#10 plasmid was also introduced to obtain the V12Sα strains where the lysE, dapA, dapB, asd, lysA, and ddh genes were introduced into the chromosome by the method described in Example 16. Randomly selected 200 strains were cultured in an SEII production medium containing 50 mg/L streptomycin and 20 mg/L kanamycin to select a strain which produced the most L-lysine, and was designated as Vmac1. A streptomycin-sensitive strain was selected form the Vmac1 strains to obtain a strain where pAET7 was eliminated. The pFLP31 plasmid shown in Example 9 was introduced into the strain, and a kanamycin-resistant gene region was eliminated from the mini-Mu cassette on the chromosome by the method described in Example 9. The kanamycin-sensitive strains obtained by the procedure were designated as Vmac1S strain. A streptomycin-sensitive strain was selected from the Vmac1S strain to obtain a strain where the pFLP31 plasmid was eliminated. The pAET7 plasmid was introduced again into this strain, and the plasmid pMIV-FRTKmFRT-EABDAS#10 was also introduced again. Then, the strain which produced the most L-lysine was selected and designated as the Vmac2 strain. In the same way as described above, the Vmac2S strain, Vmac3 strain, and Vmac3S strain were obtained. The amount of Lys produced by V12 was defined as 100, and the relative value of the amount of Lys produced by the Vmac3S strain was calculated and shown in Table 10.

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