METABOLIC ENGINEERING OF CUPRIAVIDUS NECATOR FOR IMPROVED FORMATE UTILIZATION
Disclosed herein are compositions and methods to improve Cupriavidus sp. as a host for formate conversion. Also disclosed herein are compositions and methods to improve growth of non-naturally occurring Cupriavidus sp. on carbon dioxide, succinate, formate and fructose as sole carbon sources.
1 . A non-naturally occurring Cupriavidus sp. comprising at least one genetic deletion wherein the at least one genetic deletion improves growth on formate as a sole carbon source by up to 24 percent over a naturally occurring Cupriavidus sp.
2 . The non-naturally occurring Cupriavidus sp. of claim 1 wherein the Cupriavidus sp. genotype comprises ΔhoxFUYHWI ΔhypA2B2F2.
3 . The non-naturally occurring Cupriavidus sp. of claim 1 wherein the Cupriavidus sp. genotype comprises ΔhoxKGZMLOQRTV ΔhypA1B1F1CDEX ΔhoxABCJ.
4 . The non-naturally occurring Cupriavidus sp. of claim 1 wherein the Cupriavidus sp. genotype comprises ΔcbbR′ ΔcbbLpSpXpYpEpFpPpTpZpGpKpAp.
5 . The non-naturally occurring Cupriavidus sp. of claim 1 wherein the Cupriavidus sp. genotype comprises ΔpHG1.
6 . The non-naturally occurring Cupriavidus sp. of claim 1 wherein the Cupriavidus sp. genotype comprises ΔphcA.
7 . The non-naturally occurring Cupriavidus sp. of claim 1 wherein the Cupriavidus sp. genotype comprises ΔpHG1 ΔphcA.
8 . The non-naturally occurring Cupriavidus sp. of claim 1 wherein the Cupriavidus sp. grows in minimal salt media supplemented with 50 mM sodium formate at a growth rate of up to 2.18 times greater than a wildtype Cupriavidus sp. grown in minimal salt media supplemented with 50 mM sodium formate.
9 . The non-naturally occurring Cupriavidus sp. of claim 1 wherein the Cupriavidus sp. grows in minimal salt media supplemented with 50 mM sodium formate up to a 34 percent greater optical density at 600 nm compared to a wildtype Cupriavidus sp. grown in minimal salt media supplemented with 50 mM sodium formate.
10 . The non-naturally occurring Cupriavidus sp. of claim 1 wherein the Cupriavidus sp. is Cupriavidus necator.
11 . A non-naturally occurring Cupriavidus sp. comprising at least one genetic deletion wherein the at least one genetic deletion improves growth on fructose as a sole carbon source by up to 19 percent over a naturally occurring Cupriavidus sp.
12 . The non-naturally occurring Cupriavidus sp. of claim 11 wherein the Cupriavidus sp. genotype comprises ΔpHG1.
13 . The non-naturally occurring Cupriavidus sp. of claim 11 wherein the Cupriavidus sp. genotype comprises ΔphcA.
14 . The non-naturally occurring Cupriavidus sp. of claim 11 wherein the Cupriavidus sp. genotype comprises ΔpHG1 ΔphcA.
15 . A non-naturally occurring Cupriavidus sp. comprising at least one genetic deletion wherein the at least one genetic deletion improves growth on succinate as the sole carbon source by up to 7 percent over a naturally occurring Cupriavidus sp.
16 . The non-naturally occurring Cupriavidus sp. of claim 15 wherein the Cupriavidus sp. genotype is selected from the group consisting of ΔpHG1 ΔphcA and ΔphcA.
17 . A non-naturally occurring Cupriavidus sp. comprising at least one genetic deletion wherein the at least one genetic deletion improves growth on carbon dioxide as a sole carbon source when compared to a naturally occurring Cupriavidus sp.
18 . The non-naturally occurring Cupriavidus sp. of claim 17 wherein the Cupriavidus sp. genotype comprises a deletion of at least one copy of the CBB operon.
19 . The non-naturally occurring Cupriavidus sp. of claim 17 wherein the Cupriavidus sp. genotype comprises a deletion of a CBB operon within a megaplasmid.
20 . The non-naturally occurring Cupriavidus sp. of claim 17 wherein the Cupriavidus sp. genotype comprises a deletion of a chromosomal CBB operon.
21 . A method for deleting a megaplasmid within an organism comprising deleting a gene on the megaplasmid that encodes for a toxin; and further comprising deleting a replication region of the megaplasmid.
22 . The method of claim 21 wherein the organism is a Cupriavidus sp.
23 . The method of claim 21 wherein the megaplasmid is pHG1.