Enhanced metabolism and negative regulation of ER stress support higher erythropoietin production in HEK293 cells.

Recombinant protein production can cause severe stress on cellular metabolism, resulting in limited titer and product quality. To investigate cellular and metabolic characteristics associated with these limitations, we compare HEK293 clones producing either erythropoietin (EPO) (secretory) or GFP (non-secretory) protein at different rates. Transcriptomic and functional analyses indicate significantly higher metabolism and oxidative phosphorylation in EPO producers compared with parental and GFP cells.

A gene cluster for chlorate metabolism in Ideonella dechloratans.

Chlorate reductase has been isolated from the chlorate-respiring bacterium Ideonella dechloratans, and the genes encoding the enzyme have been sequenced. The enzyme is composed of three different subunits and contains molybdopterin, iron, probably in iron-sulfur clusters, and heme b. The genes (clr) encoding chlorate reductase are arranged as clrABDC, where clrA, clrB, and clrC encode the subunits and clrD encodes a specific chaperone.

Cloning, characterisation, and expression of a novel gene encoding chlorite dismutase from Ideonella dechloratans.

The gene for chlorite dismutase was isolated from a lambda ZAP genomic library of Ideonella dechloratans using peptide sequences obtained from the purified protein. A nucleotide sequence of 1200 bp was determined. The chlorite dismutase gene codes for a 285-residue polypeptide, with the experimentally determined N-terminus of the mature protein at residue 38.

Chlorite dismutase from Ideonella dechloratans.

Chlorite dismutase has been purified from the chlorate-metabolizing bacterium Ideonella dechloratans. The purified enzyme is tetrameric, with a relative molecular mass of 25,000 for the subunit, and contains about 0.6 heme/subunit as isolated.