Expression of a Thermostable Enzyme from Thermobifida fusca in Pichia pastoris
Author Information
Author(s): Yang Chao-Hsun, Lin Kun-I, Chen Gen-Hung, Chen Yu-Fen, Chen Cheng-Yu, Chen Wei-Lin, Huang Yu-Chun
Primary Institution: Providence University
Hypothesis
Can the AXE gene from Thermobifida fusca be expressed in Pichia pastoris for enhanced enzyme production?
Conclusion
The study successfully expressed the AXE gene in Pichia pastoris, resulting in high levels of enzyme production.
Supporting Evidence
- The AXE gene was successfully cloned and expressed in Pichia pastoris.
- High levels of AXE activity (526 U/mL) were achieved in the culture broth.
- The enzyme maintained about 70% of its activity after heat treatment at 60 °C for three hours.
- The GAP expression system was found to be more suitable for large-scale production than the AOX system.
Takeaway
Scientists took a gene from a heat-loving bacteria and put it into yeast to make a special enzyme that can work at high temperatures.
Methodology
The AXE gene was amplified, cloned into a vector, and expressed in Pichia pastoris, followed by enzyme activity assays.
Limitations
The thermostability of the AXE from P. pastoris was lower than that from the original strain, T. fusca NTU22.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website