Bacterial cells with improved tolerance to polyamines
AssigneeDanmarks Tekniske Universitet
InventorsRebecca Lennen, Alex Toftgaard Nielsen, Markus Herrgård, Morten Sommer, Adam Feist, Elsayed Tharwat Tolba Mohamed
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Abstract
Provided are bacterial cells genetically modified to improve their tolerance to certain commodity chemicals, such as polyamines, and methods of preparing and using such bacterial cells for production of polyamines and other compounds.
Claims20
A bacterial cell comprising a recombinant biosynthetic pathway for producing an aliphatic polyamine and at least one genetic modification which reduces expression of an endogenous gene selected from the group consisting of proV, proW, proX, cspC, ptsP, wbbK, yobF, nagC, nagA, rph, ybeX and mpl, or a combination of any thereof.
A bacterial cell comprising at least one mutation selected from YgaC-R43L, RpsG-L157*, MreB-A298V, MreB-N34K, MreB-E212A, MreB-I24M, MreB-H93N, NusA-L152R, NusA-M204R, SspA-F83C, SspA-V91F, MrdB-E254K, RpoD-E575A, RpoC-V401G, RpoC-V453I, RpoC-R1140C, RpoC-L120P, RpoB-R637L, RpoB-G181V, MurA-G141A, MurA-Y393S, RpsA-D310Y, RpsA-D310G, RpsA-D160V, RpsA-N313K, RpsA-N315K, RpsA-E427R, SpoT-R209H, SpoT-R467H, SpoT-R471H, SpoT-R488C, SpoT-G530C and argG-C324A.
A process for improving the tolerance of an E. coli cell to at least one aliphatic polyamine selected from putrescine, HMDA, 1,3-diaminopropane, spermidine, agmatine, cadaverine, ethylenediamine, citrulline, and ornithine, comprising
- a) genetically modifying the E. coli cell to knock-down or knock-out at least one endogenous gene selected from the group consisting of proV, proW, proX, cspC, ptsP, wbbK, yobF, nagC, nagA, rph, ybeX and mpl;
- b) preparing a population of E. coli cells comprising one or more mutations in at least one endogenous gene selected from ygaC, rpsG, mreB, nusA, sspA, mrdA, rpoD, rpoC, rpoB, murA, rpsA, spoT, and argG;
- and selecting any host cell which has an improved tolerance to at least one of putrescine, HMDA, 1,3-diaminopropane, spermidine, agmatine, cadaverine, ethylenediamine, citrulline, and ornithine;
- or
- c) both a) and b).
A composition comprising putrescine, HMDA, cadaverine, spermidine, agmatine, 1,3-diaminopropane, ethylenediamine, citrulline, or ornithine at a concentration of at least 10 g/L and a plurality of bacterial cells of the Escherichia genus which comprise
- a) at least one genetic modification which reduces expression of an endogenous gene selected from the group consisting of proV, proW, proX, cspC, ptsP, wbbK, yobF, nagC, nagA, rph, ybeX and mpl, or a combination of any thereof;
- b) a mutation in at least one of ygaC, rpsG, mreB, nusA, sspA, mrdA, rpoD, rpoC, rpoB, murA, rpsA, spoT and argG which improves the tolerance of the bacterial cell to putrescine, HMDA, cadaverine, 1,3-diaminopropane, spermidine, agmatine, ethylenediamine, citrulline, or ornithine;
- or
- c) a combination of a) and b).
Dependencies are inferred from the claim text.