EC Number   |
|---|
   1.14.13.69 | expression in Xanthobacter autotrophicus |
| 1.14.13.69 | expression of wild-type and mutant enzymes |
| 1.14.13.69 | gene etnC, expression analysis under different culture conditions, overview |
| 1.14.13.69 | gene etnD, recombinant enyme expression of the gene cluster EtnABCD in Pseudomonas putida strain KT2440, cloning the EtnABCD gene cluster into Pseudomonas putida KT2440 yields detectable epoxidation of ethene, which is stimulated by the coexpression of cpn60 chaperonins from either Mycobacterium spp. or Escherichia coli, the complete EtnABCD enzyme cannot be expressed in Escherichia coli strains BL21(DE3)(pLysS) or Rosetta2 although recombinant EtnB and EtnD proteins can be purified individually in soluble form. Strong functional expression of EtnABCD is achieved in Mycobacterium smegmatis mc2155 from pUS116 plasmid, yielding high epoxidation activity. Commercially available codon-optimizing Escherichia coli strains, such as Rosetta2 (Novagen), cannot supply all of the required rare codons. Methods, overview. Gene etnH is not essential for MO activity |
| 1.14.13.69 | native and His-tagged protein expressed in Escherichia coli Rosetta(DE3)(pLysS) |
| 1.14.13.69 | recombinant functional expression of the PmoABCD gene cluster in Mycobacterium smegmatis strain mc2155 from pUS116 plasmid |
| 1.14.13.69 | the oxygenase component of IsoMO from Rhodococcus sp. strain AD45 is recombinantly expressed as functional protein in Rhodococcus sp. strain AD45-ID from plasmid pTipQC2S2:isoEx, which encodes the complete IsoMO gene cluster, including an N-terminal StrepII tag on IsoA. Recombinant expression of N-terminally His6-tagged IsoC in Escherichia coli. Recombinant expression of N-terminally StrepII-tagged IsoD in Escherichia coli strain Rosetta2/(pLysS) using plasmid pET51b:isoD. Expression of His-tagged, untagged, glutathione S-transferase (GST)-fusion and maltose binding protein (MBP)-fusion IsoF, via both homologous expression in Rhodococcus sp. AD45-ID and heterologous expression in Escherichia coli Rosetta2/(pLysS), results in protein which is either expressed entirely in inclusion bodies, expressed so poorly it is impossible to purify, or when purified does not contain the predicted cofactors or demonstrate the ability to reduce IsoC |
