ABF DFO with Technology Holding, Inc.

Gregg Beckham, Mukund Karanjikar

Research output: NRELPresentation


This Agile BioFoundry Directed Funding Opportunity project with Technology Holding and partners focuses on the development of both a strain of Pseudomonas putida KT2440 and a corresponding bioprocess to convert cellulosic sugars to beta-ketoadipic acid, which can be used in performance nylons and polyesters. Our approach follows the Design-Build-Test-Learn cycle wherein we have transferred learnings from muconic acid production in P. putida to develop a glucose and xylose-utilizing beta-ketoadipic acid production strain. This strain achieves 65 g/L of beta-ketoadipic acid at 0.7 g/L/hr and a C-mol yield of 0.40. We are currently on-boarding arabinose utilization as well. To identify non-intuitive strain modifications as well, we are deploying a beta-ketoadipic acid biosensor and building randomly barcoded transposon insertion sequencing (RB-TnSeq) libraries and gene over-expression libraries in beta-ketoadipic acid production strains. Moreover, we are using global metabolomics and other systems biology tools to identify off-target pathways. Lastly, we are scaling up beta-ketoadipic acid production to kg-scale production for Technology Holding to evaluate in performance polymers with their partners.
Original languageAmerican English
Number of pages11
StatePublished - 2023

Publication series

NamePresented at the 2023 U.S. Department of Energy's Bioenergy Technologies Office (BETO) Project Peer Review, 3-7 April 2023, Denver, Colorado

NREL Publication Number

  • NREL/PR-2A00-85695


  • beta-ketoadipic acid
  • Pseudomonas putida KT2440


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