Difference between revisions of "Part:BBa K1469002"

(→‎Improvement)
 
(10 intermediate revisions by 2 users not shown)
Line 4: Line 4:
 
UDP-glucose 6-dehydrogenase of ''Bacillus megaterium''. The enzyme catalyses the NAD dependent oxidation of UDP-glucose to UDP-glucuronic acid, a key precursor molecule for hyaluronic acid production.  
 
UDP-glucose 6-dehydrogenase of ''Bacillus megaterium''. The enzyme catalyses the NAD dependent oxidation of UDP-glucose to UDP-glucuronic acid, a key precursor molecule for hyaluronic acid production.  
  
−
<!-- Add more about the biology of this part here
 
−
===Usage and Biology===
 
  
−
<!-- -->
 
−
<span class='h3bb'>Sequence and Features</span>
 
−
<partinfo>BBa_K1469002 SequenceAndFeatures</partinfo>
 
  
−
 
+
=====Characterization by SSTi-SZGD(2018)=====
−
<!-- Uncomment this to enable Functional Parameter display
+
−
===Functional Parameters===
+
−
<partinfo>BBa_K1469002 parameters</partinfo>
+
−
<!-- -->
+
−
=====Characterization by SSTi-SZG(2018)=====
+
 
<html>
 
<html>
 
<body>
 
<body>
−
<p>This is an improvement of this part and gtaB (BBa_K1469005), which were originally introduced by Saarland iGEM in 2014. We characterized this part by co-overexpressing it with gtaB (BBa_K1469005) together in an operon. UDP-GlcDH is also known as tuaD in B.subtilis, and its gene product may contribute positively to HA production.</p><br>
+
<p>We characterized this part by co-overexpressing it with gtaB (<a href="https://parts.igem.org/Part:BBa_K1469005">BBa_K1469005</a>) together in an operon. UDP-GlcDH is also known as tuaD in B.subtilis, and its gene product may contribute positively to HA production.</p><br>
  
 
<p>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. Gene products of tuaD and gtaB regulate the last two steps in the synthetic pathway of UDP-GlcUA, one of the two precursors of HA. </p><br>
 
<p>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. Gene products of tuaD and gtaB regulate the last two steps in the synthetic pathway of UDP-GlcUA, one of the two precursors of HA. </p><br>
  
−
<img src="https://static.igem.org/mediawiki/parts/c/cb/T--SSTi-SZGD--acid_pathway.png"style="width:50%">
+
<img src="https://static.igem.org/mediawiki/parts/c/cb/T--SSTi-SZGD--acid_pathway.png"style="width:40%">
−
<img src="https://static.igem.org/mediawiki/parts/7/7d/T--SSTi-SZGD--p43nmk_acid.png" style="width:50%">
+
<img src="https://static.igem.org/mediawiki/parts/7/7d/T--SSTi-SZGD--p43nmk_acid.png" style="width:40%">
  
 
<p>Fig1  the synthesis pathway of HA .  
 
<p>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 ). </p>
 
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 ). </p>
  
−
<img src="https://static.igem.org/mediawiki/parts/3/3f/T--SSTi-SZGD--CTAB_solution.png"style="width:50%">
+
<img src="https://static.igem.org/mediawiki/parts/e/e4/T--SSTi-SZGD--CTAB_solution.jpeg"style="width:40%">
 
<p>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. </p><br>
 
<p>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. </p><br>
−
<img src="https://static.igem.org/mediawiki/parts/4/4c/T--SSTi-SZGD--viscometer_analysis.png" style="width:50%">
+
<img src="https://static.igem.org/mediawiki/parts/4/4c/T--SSTi-SZGD--viscometer_analysis.png" style="width:40%">
 
<p>Figure 3: molecular weights of HA produced by overexpression of precursor genes in recombinant B. subtilis 168E strains using viscometer analysis.  </p>
 
<p>Figure 3: molecular weights of HA produced by overexpression of precursor genes in recombinant B. subtilis 168E strains using viscometer analysis.  </p>
  
 
</body>
 
</body>
 
</html>
 
</html>
 +
 +
<!-- Add more about the biology of this part here
 +
 +
 +
 +
===Usage and Biology===
 +
 +
<!-- -->
 +
<span class='h3bb'>Sequence and Features</span>
 +
<partinfo>BBa_K1469002 SequenceAndFeatures</partinfo>
 +
 +
 +
<!-- Uncomment this to enable Functional Parameter display
 +
===Functional Parameters===
 +
<partinfo>BBa_K1469002 parameters</partinfo>
 +
<!-- -->

Latest revision as of 01:59, 18 October 2018

UDP-GlcDH

UDP-glucose 6-dehydrogenase of Bacillus megaterium. The enzyme catalyses the NAD dependent oxidation of UDP-glucose to UDP-glucuronic acid, a key precursor molecule for hyaluronic acid production.


Characterization by SSTi-SZGD(2018)

We characterized this part by co-overexpressing it with gtaB (BBa_K1469005) together in an operon. UDP-GlcDH is also known as tuaD in B.subtilis, and its 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. Gene products of tuaD and gtaB regulate the last two steps in the synthetic pathway of UDP-GlcUA, one of the two precursors of HA.


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.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]