Part:BBa_K3402055

Over-expression UGTB cassette

This device is composed of 50bp-upPXA1(BBa_K3402037), Six site(BBa_K3402032), Pgalk(BBa_K3402033), Rec(BBa_K3402034), Tgki(BBa_K3402019), hph(BBa_K3402012), Ptef1(BBa_K3402007), UGTB(BBa_K3402011), Tsyn7(BBa_K3402001), 50bp-doPXA1(BBa_K3402038).

Usage and Biology

upPXA1 and doPXA1 are homologous arms, which means this device will edit the PXA1 site. The β-Rec/six self-excising system will help to excise the hygromycin resistance gene. UDP-glucosyltransferase B (UGTB) is the key gene in the synthesis of sophorolipids.
Choose PXA1 as the gene editing site. Three strong promoters Ptef1, Peno and Pgki were knocked in PXA1 site to over-express UDP-glucosyltransferase B (UGTB).
Then we transformed the corresponding over-expression fragments and the Cas9 and sgRNA expression cassette to the wild-type Starmerella bombicola.
After incubation and fermentation, test the yield and acid/lactone ratio of sophorolipids produced by different strains. After over-expressing UGTB, the yield could be increased a lot than that of the control.

Table. 1 The yield of sophorolipids produced by different strains

Chart.1 The yield of sophorolipids by over-expression of UGTB
(W: wild type; S1: ::Ptef1-UGTB; S2: ::Peno-UGTB; S3: ::Pgki-UGTB)

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 1076
  Illegal BglII site found at 5893
  Illegal BamHI site found at 3479
  Illegal XhoI site found at 1910
  Illegal XhoI site found at 4631
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 219
  Illegal AgeI site found at 5315
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 5474
  Illegal BsaI.rc site found at 5429