Part:BBa_K3819022
Lipase plasmid
The lipase plasmid expresses lipase which can not only assist cellulase and xylanase in deinking during deinking, but also effectively treat the deinking after deinking sewage In order to ensure that the lipase can be smoothly secreted from Pichia pastoris GS115. We also introduced SMT3, the SUMO fusion sequence is used to increase the expression of Pichia pastoris lipase, and the SMT3 and lipase sequences are connected by GS linker (GGGGS) 2. after the lipase sequence, we also added Myctag as a late purification tag.
Finally, we successfully expressed this recombinant plasmid and detected the activity of lipase, which means lipase can work normally. And the results are as follows.
- Figure 1.The Dot-Blot results of recombined GS115(pLiplac) fermentation broth. The sample was gained after fermentation in YPD for 96h. Lipase(Myc-Tag) was tested by adding 2ul fermentation supernatant on NC membrane and exposing 60s.
- Figure 2. Alkali Titration results of Lipase. 1ml concentrated protein solution mixed with 5ml 50mM, PH=5.0 phosphate buffer and 4ml emulsion, then reacted at 40℃ for 15min. In the experiment system. Produced fatty acid neutralizes 100mM NaOH solution so experiment system's color was stable during titration, while system turn red directly in control.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 727
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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