Molecular cloning of the L-phenylalanine transaminase gene from Paracoccus denitrificans in Escherichia coli K-12.
The L-phenylalanine transaminase gene of Paracoccus denitrificans was cloned by a shotgun method using the Escherichia coli K-12 mutant DG30, which lacks three distinct transaminase genes. Plasmid pPAP142 was constructed by inserting a 2.2-kb fragment carrying the transaminase gene into pUC18. Strain E. coli K-12 HB101 cells harboring the plasmid produced 20-fold to 30-fold more transaminase than wild type P. denitrificans cells. The nucleotide sequence of the 2.2-kb fragment was determined, revealing that the deduced amino acid sequence of the transaminase of P. denitrificans is similar to that of other transaminases.
Takagi
T
T
Research Laboratory of Applied Biochemistry, Tanabe Seiyaku Co. Ltd, Osaka, Japan.
Taniguchi
T
T
Yamamoto
Y
Y
Shibatani
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000302:isolation & purification; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D010231:Paracoccus denitrificans / Q000201:enzymology / Q000235:genetics*; D010649:Phenylalanine / Q000096:biosynthesis; D010957:Plasmids / Q000235:genetics; D000637:Transaminases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification
D004269:DNA, Bacterial; D004274:DNA, Recombinant; D010649:Phenylalanine; D000637:Transaminases; C022340:phenylalanine aminotransferase