| Claim/Topic | Evidence summary | Organism/context | Quantitative data | Source (with year, DOI URL) |
|---|---|---|---|---|
| Core enzymatic reaction of shikimate kinase | Shikimate kinase (SK; EC 2.7.1.71) phosphorylates shikimate at the C3 hydroxyl using ATP to form shikimate-3-phosphate (S3P), a required step toward EPSP and chorismate in the shikimate pathway (pqac-00000001) | General bacterial/plant/algal shikimate-pathway context | Reaction: shikimate + ATP -> shikimate-3-phosphate + ADP | Niraula et al., 2025, https://doi.org/10.3390/biotech14010006 |
| Bacterial isoenzymes AroK and AroL | In bacteria, two shikimate kinase isoenzymes are commonly recognized: shikimate kinase I (AroK) and shikimate kinase II (AroL). Engineering studies often repress/delete these enzymes to accumulate shikimate, but full loss causes auxotrophy and necessitates aromatic supplementation (pqac-00000002) | Engineered bacterial shikimate producers, especially E. coli | Example reported for engineered E. coli: 13.15 g/L shikimate in 5-L fed-batch using tunable aroK expression rather than permanent deletion | Gu et al., 2016, https://doi.org/10.1038/srep29745 |
| Shikimate-binding determinants in AroK | Structural/functional analysis of bacterial AroK identifies shikimate-binding subsites CX, OCORE, and OLID. Conserved residues contacting the substrate include positions corresponding to R60 and R140; mutating these severely impaired function/growth in engineering contexts (pqac-00000003) | Bacterial AroK, with cited structural work including Helicobacter pylori and other bacterial SKs | No single kinetic value in snippet; severe growth inhibition observed for key conserved-site mutants | Bo et al., 2023, https://doi.org/10.3390/metabo13060747 |
| KT2440-specific aroK manipulation in muconate engineering | In Pseudomonas putida KT2440-derived strains for muconate production, aroK was deliberately overexpressed from Ptac, either alone or together with aroB (e.g., ΔpykF::Ptac:aroK and ΔpykF::Ptac:aroK:aroB), indicating aroK is a practical flux-control point in the native shikimate pathway (pqac-00000004) | Pseudomonas putida KT2440 metabolic engineering for muconic acid | Muconate process in study reached 33.7 g/L muconate, 0.18 g/L/h, 46% molar yield (92% of maximum theoretical yield); snippet specifically highlights Ptac:aroK strain designs | Ling et al., 2022, https://doi.org/10.1038/s41467-022-32296-y |
| 2024 pABA combinatorial engineering identifies pathway bottleneck | A 2024 DoE study in P. putida varied shikimate- and pABA-pathway gene expression across 14 representative strains sampled from 512 possible combinations. The analysis identified aroB, not aroK, as a significant bottleneck for pABA production, while noting aroK overexpression had been beneficial in related production contexts (pqac-00000005) | Pseudomonas putida shikimate/pABA pathway engineering | Titers ranged from 2 to 186.2 mg/L initially; second-round designs reached ~232.1 mg/L | Campos-Magaña et al., 2024, https://doi.org/10.1101/2024.06.17.599342 |
| 2024 shikimate pathway-dependent catabolism (SDC) | A 2024 P. putida study rewired metabolism so shikimate-pathway-derived reactions supplied pyruvate, demonstrating strong metabolic plasticity of the pathway relevant to enzymes such as AroK that control flux into downstream aromatic intermediates (pqac-00000006) | Pseudomonas putida engineered for aromatic production | 89% of maximum theoretical yield for 4-hydroxybenzoate in minimal medium; adapted strains also produced salicylate and 3-hydroxybenzoate at high yields | dos Santos et al., 2024, https://doi.org/10.21203/rs.3.rs-4761679/v1 |
| 2023 real-world aromatic production application in KT2440 | P. putida KT2440 was engineered to produce gallic acid from glycerol using a synthetic operon (aroG4, quiC, pobA*) plus deletions blocking degradation (pcaHG and galTAPR). This shows the practical biotechnological value of controlling shikimate-pathway flux in KT2440, even though aroK itself was not directly manipulated in the snippet (pqac-00000007) | Pseudomonas putida KT2440 producing gallic acid from glycerol | 346.7 ± 0.004 mg/L gallic acid after 72 h in shake flasks | Dias et al., 2023, https://doi.org/10.1007/s10123-022-00282-5 |


*Table: This table summarizes the most relevant gathered evidence for annotating Pseudomonas putida KT2440 aroK as shikimate kinase, including core reaction chemistry, bacterial isoenzyme context, and KT2440-specific engineering evidence. It is useful for separating direct functional evidence from broader pathway and application-level evidence.*