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. 2023 Nov 6;11:1296132. doi: 10.3389/fbioe.2023.1296132

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This work is authored by Jinho Kim and Taek Soon Lee on behalf of the U.S. Government and as regards Dr. Kim, Dr. Lee and the U.S. Government, is not subject to copyright protection in the United States. Foreign and other copyrights may apply.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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(A) A pathway map of gene knockdown targets for redirecting metabolic flux toward isoprenol production using CRISPRi. Pathways and relevant genes are colored as follows: green box, glycolysis; purple box, amino acids pathway; violet box, propionic acid pathway; light blue box, nucleotides pathway; yellow box, organic acids pathway; orange box, isoprenoids pathway; red box, TCA cycle pathway. PRA, propionic acid; PEP, phosphoenolpyruvate; TCA, citric acid cycle. (B) Original (yellow box) and IPP-bypass (red box) MVA pathways for isoprenol production. The MVA pathway converts acetyl-CoA to IPP in 6 enzymatic steps with subsequent isomerization to dimethylallyl diphosphate (DMAPP). Dephosphorylation of IPP and DMAPP by nudB, a promiscuous E. coli phosphatase, produces isoprenol and prenol, respectively. The IPP-bypass pathway was proposed in this study for direct decarboxylation of mevalonate monophosphate (MVAP) to isopentenyl monophosphate (IP) followed by dephosphorylation by an endogenous phosphatase. ADP, adenosine diphosphate; AtoB, acetoacetyl-CoA thiolase; ATP, adenosine triphosphate; DMAPP, dimethylallyl diphosphate; HMG-CoA, 3-hydroxy-3-methyl-glutaryl-CoA; HMGR, HMG-CoA reductase; HMGS, HMG-CoA synthase; IDI, isopentenyl diphosphate isomerase; IP, isopentenyl monophosphateIPP, isopentenyl diphosphate; MK, mevalonate kinase; MVA, mevalonate; MVAP, mevalonate monophosphate; MVAPP, mevalonate diphosphate; PMD, mevalonate diphosphate kinase; PMK, phosphomevalonate kinase.