Engineering of global transcription factor FruR to redirect the carbon flow in Escherichia coli for enhancing l-phenylalanine biosynthesis

Table 4 Main strains and plasmids used in this study

Strains	Characteristics	Sources
E. coli W3110MT	W3110 derived mutant, l-tyrosine auxotrophic	Our lab
PHE01	W3110MT p15A::pL-aroF-tyrA¹⁻¹²¹-pR-pheA^T326P, kanamycin resistance	Our lab
PHE02	PHE01/pCas9, spectinomycin resistance	This work
PHE03	PHE01ΔfurR	This work
PHE04	PHE01ΔaroF^MT:: aroF^WT	This work
PHE05	PHE01ΔpheA^MT:: pheA^WT	This work
PHE06	PHE01ΔfruR::Cm^R/pmtr-RFP	This work
PHE07	PHE01ΔfurR::furR^E173K	This work
PHE08	PHE01ΔfurR::furR^A18P	This work
PHE09	PHE01ΔfurR::furR^L3F−L170N	This work
PHE10	PHE01ΔfurR::furR^S75I−V160E	This work
PHE11	PHE01ΔfurR::furR^P219H	This work
PHE12	PHE01ΔfurR::furR^I144T	This work
PHE13	PHE01ΔfurR::furR^E72R−P128A	This work
PHE14	PHE01ΔfurR::furR^R312G	This work
Plasmids
pCas	expressing Cas9 protein and offering sgRNA for removing donor plasmid, Spectinomycin resistance	[33]
pGRB	plasmid for expressing sgRNA or with offering donor DNA, Ampicillin resistance	[33]
pGRB-ΔfruR	pGRB fruR-sgRNA^a	This work
pGRB-aroF^WT	pGRB aroF^MT-sgRNA ΔaroF^MT::aroF^WT	This work
pGRB-pheA^WT	pGRB pheA^MT-sgRNA ΔpheA^MT::pheA^WT	This work
pGRB-ΔfruR::Cm^R	pGRB fruR-sgRNA ΔfruR::Cm^R	This work
pGRB-fruR^MT	pGRB Cm^R-sgRNA ΔCm^R::fruR^MT	This work
pmtr-RFP	pBR332 pmtr-RFP (_L-PHE biosensor)	This work

ISSN: 1475-2859