Part:BBa_K1469005

gtaB

UTP--glucose-1-phosphate uridylyltransferase of Bacillus megaterium. The enzyme catalyzes the conversion of glucose 1-phospahate to UDP-glucose 1-phosphate.

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 768
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 220
  

Characterization by SSTi-SZGD(2018)

This is an improvement of this part and tuaD(UDP-GlcDH )(BBa_K1469002), which were originally introduced by saarland iGEM in 2014. We characterized this part by co-overexpressing it with tuaD(BBa_K1469002)together in an operon, it’s gene product may contribute positively to HA production.

In our project, expression of the operon tuaD-gtaB was regulated under the control of a constitutive promoter P43, this operon is used for further increasing the production of the HA in B.subtilis, tuaD and gtaB gene, products regulate the last two steps in the synthetic pathway of UDP-GlcUA.

Fig1 the synthesis pathway of HA . In our experiment, by conducting CTAB experiments that form turbidity from a reaction between HA and CTAB solution, the results showed a remarkable increase in HA production when co-overexpressed tuaD-gtaB together (488mg/L, a 38% increase) (Figure2),In addition, Molecular weight analysis studies showed that HAs synthesized were high molecular weight(Figure3 ).

Figure 2: CTAB analysis of HA concentraton, a. Illustration of the turbidity by mixing different source of HA with CTAB solution. b: effects of overexpressing the precursor genes on HA production in recombinant B.

Figure 3: molecular weights of HA produced by overexpression of precursor genes in recombinant B. subtilis 168E strains using viscometer analysis.