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Revision as of 20:19, 16 October 2016

Chalmers Gothenburg iGEM 2016

ACHIEVEMENTS
Project results

Synechocystis growth trials

A growth trial was performed on both the WT and JA06 strains of Synechocystis given to us to study its growth rate, Figure 1, and the acetate production, Figure 2 & Figure 3. As can be seen in Figure 1 the wild type and mutant strain JA06 of Synechocystis followed a very similar growth curve during the 600 hours long growth trial. This is good since it indicates that the added features of the mutant strain does not put any major metabolic burden on the organism.

Fig 1.OD730 over time for Synechocystis wild type and JA06 in BG-11

When measuring the samples from the growth trial by using HPLC it was shown that the wild type had no acetate production, as was expected. The JA06 strain showed acetate levels of up to 0.22 g/l when grown in regular BG-11, Figure 2, and up to 0.74 g/l when grown in BG-11 with bicarbonate, Figure 3.

Fig 2.OD730 and acetate over time for Synechocystis JA06 in BG-11
Fig 3.OD730 and acetate over time for Synechocystis JA06 in BG-11 + NaHCO3

Although the JA06 strain already showed some promising numbers for acetate production it was believed that this number could have been increased further with the genetic modifications that were planned for the organism. None of the planned modifications were however successfully performed as there were major problems with the assembly and transformation of constructs during the project.

Growth trials of production organisms on acetate

A growth trial was performed to investigate the ability of the different organisms to naturally grow on acetate in BG-11 media. The organism that was most successfully grown under the desired conditions was Y. lipolytica, Figure 4. Some of the organisms, like B. subtilis, were not expected to grow on acetate as certain modifications are needed to make it compatible. But with all the issues that were encountered with the construct assemblies and transformations there was not time to get them working.

Fig 4.Yarrowia’s OD over time when grown in BG-11 media with 5 g/l acetate.

Y. lipolytica showed a fast stable growth at 5 g/l of acetate, but was able to grow in slightly lower concentrations as well. Considering that the current highest obtained concentration of acetate was 0.74 g/l a stable co-culture between the two organisms is not quite possible yet. With some of the modifications that were planned for the organisms it is possible that the acetate production would’ve been enough for a stable growth.

Co-culture growth trial

Promoter study

The results from the promoter study is presented in the page dedicated to describing the promoter study.

Future directions