Cloning of the ATP phosphoribosyl transferase gene of Corynebacterium glutamicum and application of the gene to L-histidine production.
Corynebacterium glutamicum mutants lacking ATP phosphoribosyl transferase (PRT) were selected by complementation with the Escherichia coli PRT gene. The recombinant plasmid pCH13 carrying a wild type PRT gene from C. glutamicum T106 was obtained in one of the mutants, LH13. Transformants, LH13/pCH13 and T106/pCH13, had three times higher PRT specific activity than T106. The plasmid pCH99 specifying the PRT, which was desensitized to feedback inhibition by L-histidine fifty-fold higher than the wild type PRT, was derived from pCH13. L-Histidine productivity of C. glutamicum F81, was markedly decreased by pCH13, but increased twice by pCH99. In cultivation in jar fermentors, F81/pCH99 continued to accumulate L-histidine through fermentation and yielded to the titer of 22/5 g/liter, while F81 accumulated only 11.5 g/liter due to production retardation halfway through fermentation. Moreover, F81/pCH99 had a larger production rate than F81 even in its production phase. These results indicate that the yield improvement results from amplification of the highly desensitized PRT provided by pCH99.
Mizukami
T
T
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co, Ltd, Japan.
Hamu
A
A
Ikeda
M
M
Oka
T
T
Katsumata
R
R
D001276:ATP Phosphoribosyltransferase / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D003352:Corynebacterium / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D005798:Genes, Bacterial; D006639:Histidine / Q000096:biosynthesis*; D009154:Mutation; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism
D011994:Recombinant Proteins; D006639:Histidine; D001276:ATP Phosphoribosyltransferase