Part:BBa_K3196017

AOX1-Kozak-FLO10 pro-pelA-His tag-AOX1 Terminator

FLO10 αpro is a combined signal peptide, which enhance the enzyme activity 3 times. pelA can catalyze pectin.

Characterization

This is a four section for degrade and transfer pectin part.

Figure1. T--HUST--China--2019-FLO10-αpropelA

DNA Gel Electrophoretic

After we link FLO10 αpro and pelA successfully, we run the PCR with an intention to confirm the expression of pelA. As the figure shows, we get the genetic stripe about 1092 bp which means the PCR is successful. Electro-transformation to Pichia pastoris was all succeeded.

Figure1 Yeast Genome extraction and pcr

Enzyme Activity

The enzyme activity of the pelA group is obtained by titration and daily measurement of enzyme activity.

Figure2 Determination of pelA enzyme activity

Pectin is a hydrophilic polysaccharide substance present in the interstitial of plant cells. It is stable in acidic medium and insoluble in ethanol. It is a natural polymer compound. We understand that pectinase measures enzyme activity has a nationally standardized experimental procedure, so we have adopted the standard titration scheme given by the state. ^[3]

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1986
  Illegal BamHI site found at 937
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]