Escherichia coliΔ¹-pyrroline-5-carboxylate reductase: gene sequence, protein overproduction and purification

Abstract

The sequence of the Escherichia coli proC gene which encodes for delta 1-pyrroline-5-carboxylate (PCA) reductase was determined. Overproduction of the proC gene product via an expression plasmid carrying the bacteriophage lambda PL promoter allowed the purification to homogeneity of PCA reductase by affinity adsorption chromatography. NH2 and COOH-terminal analysis and amino acid composition of the purified proC protein is consistent with the gene sequence reported. The molecular weight of the proC monomer is 28,112.

Molecular Cloning and Evidence for Osmoregulation of the Δ1-Pyrroline-5-Carboxylate Reductase (proC) Gene in Pea (Pisum sativum L.)

“…Second, the major ORF of the pea P5CR gene encodes a polypeptide with a predicted Mr of 28,242, and the soybean nodule P5CR antibody recognizes an Mr 28,000 polypeptide from pea tissues on SDS-PAGE immunoblots. Finally, the predicted size for the pea enzyme is also similar to that for P5CRs purified from other plant species (7,19) and from E. coli (13).…”

Osmoregulation of a Pyrroline-5-Carboxylate Reductase Gene in Arabidopsis thaliana

“…To investigate further whether Pro accumulation is a consequence of de novo transcription of genes in the biosynthetic pathway, AT-PSCI, a P5C reductase gene in A. thaliana, was cloned. Degenerate primers were chosen from amino acid stretches that were conserved among the deduced P5C reductase sequences of E. coli (Deutch et al, 1982), soybean (Delauney and Verma, 1990), and yeast (M.C. Brandriss, GenBank accession No.…”

“…A, A. thaliana; E, E. coli (Deutch et al, 1982); H, H. sap/ens (Dougherty et al, 1992); M, M. smithii (Hamilton and Reeve, 1985); P, P. aeruginosa (Savioz et al, 1990); S, G. max (Delauney and Verma, 1990) t e a a a a a a a a a a a a a a a c a a a a a a 6 6 a a a a a a a a a a a a a a a a a …”

“…Comparison of the deduced P5C reductase amino acid sequence of AT-P5C1 with that of E. coli (Deutch et al, 1982), soybean (Delauney and Verma, 1990), human (Dougherty et al, 1992), P. aeruginosa (Savioz et al, 1990), S. cerevisiae (M.C. Brandriss; GenBank No.…”

Overexpression of [delta]-Pyrroline-5-Carboxylate Synthetase Increases Proline Production and Confers Osmotolerance in Transgenic Plants

“…GSA is spontaneously cyclized to P5C, which is reduced to Pro by P5CR (encoded by the proC gene). The E. coli proBA and proC loci have been cloned and sequenced (Deutch et al, 1982(Deutch et al, ,1984. In animals and higher plants, Pro is synthesized not only from glutamate but also from Orn (Bryan, 1990;.…”