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KMID : 0578319910010020157
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Molecules and Cells
1991 Volume.1 No. 2 p.157 ~ p.162
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Overexpression of E. coli proBA Genes in Azosprifillum and Enhanced Osmotolerance
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Cho, Moo Je
Shin, Gyun Shin/Choi, Young Ju/Ga, Sang Wan/Woon, Hae Won
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Abstract
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The proB(pro-74) mutant gene codes for a mutant y-glutamyl kinase which is less sensitive to the proline feedback inhibition than the wild type in proline biosynthetic pathway. The mutant operon proBA(pro-74) was cloned into pRK290, a derivative of plasmid RK2 (IncPl) which is stable in a wide range of hosts including Azospirillum. Two fragments of 48 kb and 10.6 kb containing the mutant operon were cloned each in two orientations. The recombinant plasmids containing pro genes, named pCMK25a, pCMK25b, pCMK30a and pCMK30b, were introduced into Azospirillum lipoferum KY6 by conjugation with E. soli SM10 harboring the plasmids. The A. lipoferum KY6(pCMK25a) transconjugant produced proline about 10 folds in the absence of NaCl and 25 folds in the presence of 0.4 M NaCI than the A. lipoferum KY6 wild-type did. The inhibition of nitrogenase activity by NaCI was partially overcome by the proline overproduction by the introduced proBA(pro-74) genes in A. lipoferum KY6.
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