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Molecular cloning of the L‐phenylalanine transaminase gene from Paracoccus denitrificans in Escherichia coli K‐12
Author(s) -
Takagi T.,
Taniguchi T.,
Yamamoto Y.,
Shibatani T.
Publication year - 1991
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1991.tb00146.x
Subject(s) - paracoccus denitrificans , escherichia coli , biology , transaminase , molecular cloning , plasmid , microbiology and biotechnology , nucleic acid sequence , aspartate transaminase , gene , peptide sequence , biochemistry , enzyme , alkaline phosphatase
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.