Abstract
The gene from Xanthomonas campestris pv. phaseoli for glutamate 1-semialdehyde (GSA) aminomutase, which is involved in the C5 pathway for synthesis of δ-aminolevulinic acid (ALA), was cloned onto a multicopy plasmid, pUC18, by the complementation of an ALA-deficient mutant (hemL) of Escherichia coli. Subcloning of deletion fragments from the initial 3.5-kb chromosomal fragment allowed the isolation of a 1.7-kb fragment which could complement the hemL mutation. Nucleotide sequence analysis of the 1.7-kb DNA fragment revealed an open reading frame (ORF) that is located downstream from a potential promoter sequence and a ribosome-binding site. The ORF encodes a polypeptide of 429 amino acid residues, and the deduced molecular mass of this polypeptide is 45,043 Da. The amino acid sequence shows a high degree of homology to the HemL proteins from other organisms, and a putative binding site for pyridoxal 5′-phosphate is conserved.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Avissar YJ, Ormerod JG, Beale SI (1989) Distribution of δ-aminolevulinic acid biosynthetic pathways among phototropic bacterial groups. Arch Microbiol 151:513–519
Beale SI, Castelfranco PA (1974) The biosynthesis of δ-aminolevulinic acid in higher plants. II. Formation of 14C δ-aminolevulinic acid from labeled precursors in greening plant tissues. Plant Physiol 53:297–303
Chou PY, Fasman GD (1978) Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol 47:45–148
Drolet M, Peloguin L, Escelard Y, Cousineau L, Sasarman A (1989) Isolation and nucleotide sequence of the hemA gene of Escherichia coli K-12. Mol Gen Genet 216:347–352
Elliott T, Avissar YJ, Rhie G-E, Beale SI (1990) Cloning and sequenced the Salmonella typhimurium hemL gene and identification of the missing enzyme in hemL mutants as glutamate 1-semialdehyde aminotransferase. J Bacteriol 172:7071–7084
Ford GC, Eichele G, Jansonius JN (1980) Three-dimensional structure of a pyridoxal-phosphate-dependent enzyme, mitochondrial aspartate aminotransferse. Proc Natl Acad Sci USA 77:2559–2563
Granick S, Beale SI (1978) Hemes, chlorophylls, and related compounds: biosynthesis and metabolic regulation. Adv Enzymol 46:33–203
Grimm B (1990) Primary structure of a key enzyme in plant tetrapyrrole synthesis: glutamate 1-semialdehyde aminotransferase. Proc Natl Acad Sci USA 87:4169–4173
Grimm B, Bull A, Breu V (1991) Structural genes of glutamate 1-semialdehyde aminotransferase for porphyrin synthesis in a cyanobacterium and Escherichia coli. Mol Gen Genet 225:1–10
Ikemi M, Murakami K, Hashimoto M, Murooka Y (1992) Cloning and characterization of genes involved in the biosynthesis of δ-aminolevulinic acid in Escherichia coli. Gene 59 (I) in press
Li JM, Russel CS, Cosloy D (1989) Cloning and structure of the hemA gene of Escherichia coli K-12. Gene 82:209–217
Maniatis T, Fritsch EF, Sambrook J (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N. Y.
Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J Mol Biol 3:208–218
Murooka Y, Iwamoto H, Hamamoto A, Yamauchi T (1987) Efficient transformation of phytopathogenic strains of Xanthomonous species. J Bacteriol 169:4406–4409
O'Neill GP, Soll D (1990) Transfer RNA and the formation of a heme and chlorophyll precursor, 5-aminolevulinic acid. Biofactors 2:227–234
Sanger F, Nickelson S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Verkamp E, Chelm BK (1989) Isolation, nucleotide sequence, and preliminary characterization of the Escherichia coli K-12 hemA gene. J Bacteriol 171:4728–4735
Williams DH (1980) Black rot; a continuing threat to world crucifers. Plant Dis 64:736–742
Yanisch-Perron c, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13mp18 and pUC19 vectors. Gene 33:103–119
Author information
Authors and Affiliations
Additional information
Correspondence to: Y. Murooka
Rights and permissions
About this article
Cite this article
Murakami, K., Korbsrisate, S., Asahara, N. et al. Cloning and characterization of the glutamate 1-semialdehyde aminomutase gene from Xanthomonas campestris pv. phaseoli . Appl Microbiol Biotechnol 38, 502–506 (1993). https://doi.org/10.1007/BF00242945
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00242945