Team Meeting 8
| Date | |
|---|---|
| Department | General |
| Description | Finalising Compound Round 1 |
| HP sub-branch | |
| Links/media | |
| Participants | |
| Property | |
| Property 1 | |
| Property 2 | |
| Type | Meetings |
- Ethanol and butanol - need too many modifications - butanol yield isn't very good, ethanol medium is expensive. Methods were very discrete, different methods were used. But butanol strains we can get from yazdani. Strain - MG1655
- 1,3 propanediol - microaerobic and aerobic settings, usually produced from glycerol, 1 paper did glucose to glycerol to propanediol and 1 was a new pathway (0.6 g/l in 72 h from 20 g of glucose) Microaerobic conditions - 0.7 mol per mol of glucose, for best strain - 2.1 g/l yield. Too many modifications - 6 knockouts + 2 plasmids for microaerobic paper. Main disadvantage - main use is for non-biodegradable plastic. Minimum - 3 knockouts (microaerobic) - 0.5 g/l. Strain - Bl21 (DE3). Will produce a lot of glycerol too.
- 2 methyl butanol - potential use - biofuel (hasn't been realised yet) - 8 overexpressions, 6 knockouts. Base strain - K12. yield comparable to succinate yields, has been directly produced in PCC 7942 too and yeast. Also used for flavoring agents and pesticides.
- 2,5 furan dicarboxylic acid, have to use pseudomonas putida and ecoli. don't engineer psuedomonas, put 1 gene in e.coli, 100% conversion in 140 minutes. Add horseradish peroxidase, and catalase externally. 2 setups required - cyano + e.coli and culture pseudomonas separately. (sucrose to HMF not known). Take ecoli's supernatant, and put into psuedomonas → higher yields. requires highly acidic conditions to extract.
- glucaric acid - sucrose → glucaric acid, sucrose into 6 fructose and glucose. Fructose is used for cell growth and energy, divert glucose into glucaric acid. 3 knockouts, 1 overexpression, 2 plasmids. yields - 1.2g/l. used in biodegradable cleaners, adipic acid, corrosion inhibitors. currently produced by chemical oxidation. Aerobic conditions. E.coli strain - BL21 (DE3)
- Sorbitol - 2 papers, lactobacillus, sorbitol 6 phosphate dehydrogenase enzyme required to convert fuctose 6 phosphate to sorbitol. In lactobacillus need to overexpress srld operon (2 of em required), and knockout lactate dehydrogenase. Anaerobic conditions. overexpression of both operons was done separately, not together. ph - 7, 30-35 C. Uses - food industry, pharma. theoretical yield is 67% of glucose (resting culture). higher yields were from maltose as sugar source. sucrose also gives good yield. can't do continuous culture, batch fermentation required.
- Zymomonas - no modifs, 10% toluene treatment - glucose → ethanol + fructose → sorbitol
- Succinic acid - anaerobic pathways - 4 knockouts + 2 plasmids (pepc and citrate synthase), excluding sucrose modifs. Mg1655 strain. yield - 3.3 mol per mol of sucrose. AFp111 has 5 modifications. + pyc overexpression.
- Lactate - anaerobic pathways, pH 7, temp 37C, 2/4/6 modifications, some have adaptive evolution
Finalised compounds:
Glucaric Acid, Succinate, Lactate, Sorbitol