Phosphite synthetic auxotrophy as an effective biocontainment strategy for the industrial chassis Pseudomonas putida.
Biocontainment
NADH
Non-sterile conditions
Phosphite
Pseudomonas putida
Synthetic auxotrophy
Journal
Microbial cell factories
ISSN: 1475-2859
Titre abrégé: Microb Cell Fact
Pays: England
ID NLM: 101139812
Informations de publication
Date de publication:
08 Aug 2022
08 Aug 2022
Historique:
received:
09
03
2022
accepted:
26
07
2022
entrez:
7
8
2022
pubmed:
8
8
2022
medline:
10
8
2022
Statut:
epublish
Résumé
The inclusion of biosafety strategies into strain engineering pipelines is crucial for safe-by-design biobased processes. This in turn might enable a more rapid regulatory acceptance of bioengineered organisms in both industrial and environmental applications. For this reason, we equipped the industrially relevant microbial chassis Pseudomonas putida KT2440 with an effective biocontainment strategy based on a synthetic dependency on phosphite, which is generally not readily available in the environment. The produced PSAG-9 strain was first engineered to assimilate phosphite through the genome-integration of a phosphite dehydrogenase and a phosphite-specific transport complex. Subsequently, to deter the strain from growing on naturally assimilated phosphate, all native genes related to its transport were identified and deleted generating a strain unable to grow on media containing any phosphorous source other than phosphite. PSAG-9 exhibited fitness levels with phosphite similar to those of the wild type with phosphate, and low levels of escape frequency. Beyond biosafety, this strategy endowed P. putida with the capacity to be cultured under non-sterile conditions using phosphite as the sole phosphorous source with a reduced risk of contamination by other microbes, while displaying enhanced NADH regenerative capacity. These industrially beneficial features complement the metabolic advantages for which this species is known for, thereby strengthening it as a synthetic biology chassis with potential uses in industry, with suitability towards environmental release.
Identifiants
pubmed: 35934698
doi: 10.1186/s12934-022-01883-5
pii: 10.1186/s12934-022-01883-5
pmc: PMC9358898
doi:
Substances chimiques
Phosphates
0
Phosphites
0
Phosphorus
27YLU75U4W
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
156Subventions
Organisme : Nederlandse Organisatie voor Wetenschappelijk Onderzoek
ID : 15814
Informations de copyright
© 2022. The Author(s).
Références
J Bacteriol. 1998 Nov;180(21):5547-58
pubmed: 9791102
J Biosci Bioeng. 1999;87(3):273-9
pubmed: 16232467
Curr Protoc Mol Biol. 2007 Jul;Chapter 1:1.17.1-1.17.8
pubmed: 18265391
J Bacteriol. 1993 Jun;175(11):3430-42
pubmed: 8388873
ACS Synth Biol. 2018 Sep 21;7(9):2189-2198
pubmed: 30203964
Microbiology (Reading). 1996 Jan;142(1):155-163
pubmed: 33657738
Annu Rev Microbiol. 2007;61:379-400
pubmed: 18035609
J Microbiol Methods. 2006 Mar;64(3):391-7
pubmed: 15987659
Microb Cell Fact. 2019 Oct 22;18(1):179
pubmed: 31640713
Front Microbiol. 2015 Jul 29;6:742
pubmed: 26284036
Environ Microbiol. 2002 Dec;4(12):799-808
pubmed: 12534463
J Biol Eng. 2018 Aug 14;12:13
pubmed: 30123321
FEBS J. 2008 Oct;275(20):5173-90
pubmed: 18803670
Curr Opin Biotechnol. 2016 Apr;38:90-6
pubmed: 26874261
J Biosci Bioeng. 2012 Apr;113(4):445-50
pubmed: 22197497
Nat Protoc. 2014 Oct;9(10):2301-16
pubmed: 25188632
Science. 2003 Aug 1;301(5633):616-20
pubmed: 12893936
Front Microbiol. 2019 May 17;10:1025
pubmed: 31156576
Res Microbiol. 2004 Oct;155(8):623-9
pubmed: 15380549
Nat Commun. 2017 Nov 23;8(1):1746
pubmed: 29170493
ACS Synth Biol. 2022 Jan 21;11(1):135-143
pubmed: 34979802
Microb Biotechnol. 2021 Nov;14(6):2643-2658
pubmed: 33783967
Annu Rev Microbiol. 2002;56:743-68
pubmed: 12142492
Environ Microbiol. 2016 Oct;18(10):3535-3549
pubmed: 27233093
Microbiology (Reading). 2021 Feb;167(2):
pubmed: 33346729
Microb Biotechnol. 2019 Sep;12(5):845-848
pubmed: 31199068
Metab Eng. 2018 Nov;50:142-155
pubmed: 29758287
Front Bioeng Biotechnol. 2021 Apr 30;9:598087
pubmed: 33996776
Nat Rev Microbiol. 2014 May;12(5):368-79
pubmed: 24736795
PeerJ. 2015 Sep 15;3:e1247
pubmed: 26401457
Biotechnol J. 2021 Mar;16(3):e2000165
pubmed: 33085217
J Bacteriol. 1978 Jan;133(1):85-90
pubmed: 618850
Microb Biotechnol. 2020 Nov;13(6):1793-1806
pubmed: 32710525
Sci Rep. 2017 Mar 20;7:44748
pubmed: 28317852
ACS Synth Biol. 2015 Dec 18;4(12):1279-86
pubmed: 26072987
Microb Cell Fact. 2013 May 20;12:50
pubmed: 23687963
Appl Microbiol Biotechnol. 2012 Mar;93(6):2279-90
pubmed: 22350258
J Bacteriol. 2001 Sep;183(17):5008-14
pubmed: 11489853
Nature. 1951 Jul 21;168(4264):115-6
pubmed: 14852978
Metab Eng Commun. 2020 Mar 19;10:e00126
pubmed: 32215253
ACS Synth Biol. 2021 Oct 15;10(10):2672-2688
pubmed: 34547891
EMBO Rep. 2021 Jan 7;22(1):e51227
pubmed: 33369847
Nanoethics. 2017;11(3):297-306
pubmed: 29238409
ACS Synth Biol. 2020 Sep 18;9(9):2537-2545
pubmed: 32786356
Science. 2016 Aug 5;353(6299):583-6
pubmed: 27493184
N Biotechnol. 2021 Jan 25;60:44-51
pubmed: 32889152
BMC Microbiol. 2009 Dec 10;9:254
pubmed: 20003273
J Cell Biochem. 1993 Jan;51(1):47-54
pubmed: 8432742
ACS Synth Biol. 2016 Dec 16;5(12):1328-1330
pubmed: 27665861
Microb Biotechnol. 2020 Jan;13(1):233-249
pubmed: 30861315
Environ Microbiol. 2004 Dec;6(12):1264-86
pubmed: 15560824
BMC Microbiol. 2007 Dec 18;7:114
pubmed: 18088430
J Bacteriol. 2003 Jul;185(13):3863-70
pubmed: 12813080
Environ Microbiol. 2016 Oct;18(10):3403-3424
pubmed: 26913973
FEMS Microbiol Lett. 2011 Jul;320(1):25-32
pubmed: 21488939
Environ Microbiol. 2013 Oct;15(10):2658-71
pubmed: 23530902
Microb Biotechnol. 2009 Mar;2(2):287-94
pubmed: 21261922
Biosystems. 2019 Nov;185:104015
pubmed: 31408698
FEMS Microbiol Rev. 2010 Jul;34(4):476-95
pubmed: 20236330
Toxicol Sci. 2012 Jun;127(2):592-608
pubmed: 22454430
Microb Cell Fact. 2014 Nov 11;13:159
pubmed: 25384394
Metab Eng. 2018 Sep;49:257-266
pubmed: 30172686
ACS Omega. 2021 Apr 07;6(15):10160-10167
pubmed: 34056170
Methods Enzymol. 2013;529:329-36
pubmed: 24011059
J Bacteriol. 2004 Jul;186(14):4730-9
pubmed: 15231805
Curr Opin Biotechnol. 2018 Apr;50:195-205
pubmed: 29453021
Biotechnol Adv. 2021 Jul-Aug;49:107732
pubmed: 33785373
J Bacteriol. 1974 Sep;119(3):736-47
pubmed: 4604283
Trends Biotechnol. 2020 Dec;38(12):1308-1312
pubmed: 32402415