Engineering Pseudomonas putida for efficient aromatic conversion to bioproduct using high throughput screening in a bioreactor

Tuning a high performing multiplexed-CRISPRi Pseudomonas putida strain to further enhance indigoidine production

“…S10 ). RB-TnSeq analysis previously indicated that no transposon mutants had been recovered in either of these gene loci, which provides indirect evidence that the genes may be essential ( Eng et al, 2021 ; Price et al, 2018 ; Thompson et al, 2019 ). However, colonies were obtained for each deletion, although the strains exhibited a severe growth defect in liquid minimal medium.…”

Optimizing a High Performing Multiplex-CRISPRi P. putida Strain with Integrated Metabolomics and 13C-Metabolic Flux Analyses

“…In contrast, deletion of mqo-III (∆PP_0751) or the succinyl-CoA synthetase subunits (∆PP_4185 ∆PP_4186) led to strains with substantial growth defects (reaching OD600 ~1-3 at 24 h) and had no improvement in production titers. RB-TnSeq analysis had previously indicated that no transposon mutants had been recovered in either of these gene loci (Eng et al, 2021;Price et al, 2018;Thompson et al, 2019). Colonies were obtained for each deletion, although in liquid media the strains exhibited a several growth defect in minimal medium.…”

Engineering Pseudomonas putida KT2440 for chain length tailored free fatty acid and oleochemical production

“…Pseudomonas putida KT2440 is a promising host for industrial oleochemical production due to its remarkable ability to utilize a wide variety of carbon sources found in lignocellulosic biomass 11 , high tolerance to oxidative stress 11 , strong redox potential 12 , and favorable growth characteristics 13 . Many genetic tools have become available for engineering P. putida [14][15][16] , and significant progress has been made in improving its host properties, such as enhanced fitness in bioreactors 17 , utilization of plant-derived hydrolysates 18 , and tolerance for ionic liquids found in these hydrolysates 19 . Although P. putida has shown itself to be a versatile and robust host for bioproduction 17,[20][21][22] , to date there has not been a reported strain capable of producing free fatty acids (FFAs), which are important intermediates in oleochemical metabolic pathways as well as valuable precursors for industrial processes 23 .…”

“…Many genetic tools have become available for engineering P. putida 14–16 , and significant progress has been made in improving its host properties, such as enhanced fitness in bioreactors 17 , utilization of plant-derived hydrolysates 18 , and tolerance for ionic liquids found in these hydrolysates 19 . Although P. putida has shown itself to be a versatile and robust host for bioproduction 17,20–22 , to date there has not been a reported strain capable of producing free fatty acids (FFAs), which are important intermediates in oleochemical metabolic pathways as well as valuable precursors for industrial processes 23 .…”