{"doi":"10.1101/2020.07.27.222588","title":"Dynamic control over feedback regulatory mechanisms improves NADPH fluxes and xylitol biosynthesis in engineered <i>E. coli</i>","abstract":"Abstract We report improved NADPH flux and xylitol biosynthesis in engineered E. coli . Xylitol is produced from xylose via an NADPH dependent reductase. We utilize two-stage dynamic metabolic control to compare two approaches to optimize xylitol biosynthesis, a stoichiometric approach, wherein competitive fluxes are decreased, and a regulatory approach wherein the levels of key regulatory metabolites are reduced. The stoichiometric and regulatory approaches lead to a 16 fold and 100 fold improvement in xylitol production, respectively. Strains with reduced levels of enoyl-ACP reductase and glucose-6-phosphate dehydrogenase, led to altered metabolite pools resulting in the activation of the membrane bound transhydrogenase and a new NADPH generation pathway, namely pyruvate ferredoxin oxidoreductase coupled with NADPH dependent ferredoxin reductase, leading to increased NADPH fluxes, despite a reduction in NADPH pools. These strains produced titers of 200 g/L of xylitol from xylose at 86% of theoretical yield in instrumented bioreactors. We expect dynamic control over enoyl-ACP reductase and glucose-6-phosphate dehydrogenase to broadly enable improved NADPH dependent bioconversions. Highlights Decreases in NADPH pools lead to increased NADPH fluxes Pyruvate ferredoxin oxidoreductase coupled with NADPH-ferredoxin reductase improves NADPH production in vivo . Dynamic reduction in acyl-ACP/CoA pools alleviate inhibition of membrane bound transhydrogenase and improve NADPH flux Xylitol titers &gt; 200g/L in fed batch fermentations with xylose as a sole feedstock.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2020,"id":122902,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9515,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":564839,"name":"Eirik A. Moreb","orcid":"0000-0002-8235-494X","position":1,"is_corresponding":false},{"id":565497,"name":"Zhixia Ye","orcid":null,"position":2,"is_corresponding":false},{"id":565498,"name":"Jennifer N. Hennigan","orcid":null,"position":3,"is_corresponding":false},{"id":564840,"name":"Daniel Báez Castellanos","orcid":"0000-0003-1450-2653","position":4,"is_corresponding":false},{"id":564841,"name":"Tian Yang","orcid":"0000-0001-6923-4380","position":5,"is_corresponding":false},{"id":464906,"name":"Michael Lynch","orcid":"0000-0002-8858-9651","position":6,"is_corresponding":false},{"id":564838,"name":"Shuai Li","orcid":"0000-0002-1537-3956","position":0,"is_corresponding":true}],"reference_count":61,"raw_metadata":null,"created_at":"2026-07-18T23:14:55.385653Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}