Table 3.
Effect of CRISPR activation of the up-regulated genes on growth and production of pinene in E. coli BW25113(PT5-dxs).
| Gene | Protein | Ratio of growtha | Ratio of pinene concentrationb |
|---|---|---|---|
| Carbohydrate metabolism | |||
| ulaE | l-xylulose 5-phosphate 3-epimerase | 1.24 ± 0.01 | 1.06 ± 0.02 |
| Metabolism of terpenoids and polyketides | |||
| ispH | 4-hydroxy-3-methylbut-2-enyl diphosphate reductase, 4Fe-4S protein | 1.07 ± 0.01 | 1.19 ± 0.01 |
| dxr | 1-deoxy-d-xylulose 5-phosphate reductoisomerase | – | 1.34 ± 0.02 |
| dxs | 1-deoxyxylulose-5-phosphate synthase, thiamine triphosphate-binding, FAD-requiring | – | 1.39 ± 0.02 |
| ispU | 1-deoxy-d-xylulose 5-phosphate reductoisomerase/undecaprenyl pyrophosphate synthase | – | 1.28 ± 0.01 |
| Genetic Information Processing | |||
| rpoA | RNA polymerase, alpha subunit | 1.50 ± 0.01 | 1.09 ± 0.01 |
| dosP | oxygen sensor, c-di-GMP phosphodiesterase, | 1.56 ± 0.02 | 1.07 ± 0.01 |
| mutS | methyl-directed mismatch repair protein | 1.10 ± 0.01 | 1.29 ± 0.01 |
| cbpA | DnaK co-chaperone; curved DNA-binding protein/uncharacterized protein | 1.30 ± 0.01 | 1.04 ± 0.01 |
| dusB | methyltransferase for 50S ribosomal subunit protein L11/tRNA-dihydrouridine synthase B | 1.46 ± 0.02 | 1.23 ± 0.01 |
| Membrane protein transport Processing | |||
| gsiA | glutathione ABC transporter ATPase | 1.09 ± 0.01 | 1.05 ± 0.01 |
| nlpA | cytoplasmic membrane lipoprotein-28 | 1.64 ± 0.02 | 1.03 ± 0.02 |
| sufBCDS | SufBCD Fe-S cluster assembly protein | 1.48 ± 0.02 | 1.32 ± 0.01 |
| opgB | OPG periplasmic biosynthetic phosphoglycerol transferases I | 1.10 ± 0.01 | 1.06 ± 0.01 |
| setC | putative arabinose efflux transporter | 1.14 ± 0.02 | 1.10 ± 0.01 |
| oppF | oligopeptide ABC transporter ATPase | 1.14 ± 0.01 | 1.13 ± 0.01 |
| ypjK | CP4-57 prophage; putative inner membrane protein | 1.16 ± 0.01 | 1.10 ± 0.02 |
| btuB | vitamin B12/cobalamin outer membrane transporter | 1.05 ± 0.02 | 1.11 ± 0.01 |
| pitA | phosphate transporter, low-affinity; tellurite importer | 1.15 ± 0.01 | 1.17 ± 0.01 |
| cusA | copper/silver efflux system, membrane component | 1.15 ± 0.01 | 1.05 ± 0.01 |
| Cellular Processes | |||
| yehA | putative fimbrial-like adhesin protein | 1.48 ± 0.03 | 1.24 ± 0.02 |
| motB | protein that enables flagellar motor rotation | 1.17 ± 0.02 | 1.16 ± 0.01 |
| flgFGH | flagellar protein | 1.56 ± 0.03 | 1.41 ± 0.03 |
| ypjA | adhesin-like autotransporter | 1.12 ± 0.02 | 1.07 ± 0.01 |
| tabA | transcriptional repressor for divergent bdcA/biofilm modulator regulated by toxins; DUF386 family protein, cupin superfamily protein | 1.05 ± 0.01 | 1.05 ± 0.01 |
| Stress response | |||
| hslU | molecular chaperone and ATPase component of HslUV protease | 1.53 ± 0.03 | 1.29 ± 0.02 |
| flgFGH-sufBCDS- dusB-rpoA-yehA-hslU | 3.55 ± 0.03 | 1.71 ± 0.02 | |
a
The ratio of OD600 with CRISPRa and without.
b
The ratio of pinene production with CRISPRa and without.-: means no change or negative effect.