Team:Uppsala Universitet/Part Collection
Parts Collection
This parts collection serves as a toolbox for the expression of lignin-degrading enzymes in Pichia pastoris. Their cloning in the pPICZαB-vector places the α-secretion factor directly upstream to the N-terminus of the respective coding sequence. Therefore, desired enzymes will be secreted in the supernatant when expressed. Also, a 2A self-cleaving peptide is utilised to coexpress two proteins from one mRNA. This can be done either to express two lignin-degrading enzymes in one culture or to easily evaluate enzyme production by means of an added reporter gene like eGFP.
The purpose of this collection is to enable future iGEM teams to effectively express lignin degrading enzymes in Pichia pastoris. Furthermore, these BioBricks can be used to assess the efficiency of various enzyme mixtures to degrade lignin. A major application of the system is that it can be generalised for other enzymes that need to be produced in a eukaryotic expression system.
| # | Name | Type | Description |
|---|---|---|---|
| 1 | BBa_K3105668 | Basic Part | Horseradish peroxidase |
| 2 | BBa_K3105667 | Basic part | Aryl-alcohol oxidase |
| 3 | BBa_K3105684 | Composite part | Expression construct HRP |
| 4 | BBa_K3105685 | Composite part | Expression construct AAO |
| 5 | BBa_K3105683 | Composite part | Expression construct Glox |
| 6 | BBa_K3105682 | Composite part | Expression construct MnP |
| 7 | BBa_K3105669 | Basic part | F2A |
| 8 | BBa_K3105670 | Basic part | eGFP |
| 9 | BBa_K3105677 | Composite part | Expression construct HRP-2A-eGFP |
| 10 | BBa_K3105680 | Composite part | Expression construct HRP-2A-AAO |