Results

Biosynthesis and Optimization of 1,8-Cineole Production in Yeast

Verifying Transformation by Yeast Colony PCR and Gene Sequencing

Colony PCR verification
Gene sequencing results

Step 1: Synthesis of 1,8-Cineole

The gene of 1,8-cineole synthase was transferred into Escherichia coli, and then E. coli was transformed into yeast and colony culture was carried out.

Electrophoresis gel

Markers: 1000kb plus | 100bp

Electrophoresis Analysis

Our electrophoresis map shows the 1,8-cineole synthase gene with a gene fragment size of 1116bp. This is because it is placed between the Hindâ…¢ and EcoR1 sites of pYES2, with connectors added at both ends. The total length is 1128bp, corresponding to the positions of our two markers (100bp and 1kb plus) above the 1000 band.

Yeast colony culture
Colony selection

Step 2: Increasing the Production of 1,8-Cineole

We increased the yield of 1,8-cineole by replacing ordinary yeast cells with ERG20ww mutants and conducting SD-Ura screening.

Conclusion

We successfully obtained 1,8-cineole through biosynthesis and increased its yield using ERG20ww mutants.