Difference between revisions of "Part:BBa K3585003"

 
 
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Continuous expression of butyrate synthesis pathway enzymes through a constitutive promoter, so the strain can degrade galactose and synthesize butyrate.
 
Continuous expression of butyrate synthesis pathway enzymes through a constitutive promoter, so the strain can degrade galactose and synthesize butyrate.
  
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[[File:T--Shanghai_HS_United-BBa_K3585003 fig 00.png|500px|thumb|center|]]
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
 
 
<partinfo>BBa_K3585003 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3585003 SequenceAndFeatures</partinfo>
  
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===Contribution===
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BBa_K3585003 is a composite part consisting of butyrate synthesis pathway enzymes under constitutive promoter (basic parts BBa_J23100). The butyrate biosynthesis gene cluster can degrade galactose and synthesize butyrate.
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===Engineering success===
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The BBa_K3585003 was designed to transform galactose to butyrate, that is beneficial for galactosemia patients. In this process, the transformation of galactose to acetic acid is a crucial stage.
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Following are the results of our functional tests of BBa_K3585003 in transforming galactose to acetic acid.
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[[File:T--Shanghai HS United BBa K3585003 GC figure.png|500px|thumb|center|figure 1]]
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                                            Figure 1 Chromatographic Peak of Acetic Acid
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[[File:T--Shanghai_HS_United-BBa_K3585003_acetic_acid_new.png|500px|thumb|center|figure 2]]
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                                      Figure 2 the Concentration of Acetic Acid while Fermentation
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Each of the strains we have a repeat. Gal represents that we use galactose as the carbon source while fermentation; GG represents that we use both glucose and galactose as the carbon source while fermentation; 1917 represents wild type strains; pButy represents that the host strain contained pMTL83151-J23100-butyrate plasmids and expressed butyrate synthesis gene cluster; placY represents that the host strain contained p15A-J23200-lacY plasmids and expressed lacY gene; 1 and 2 represent for sample repeats. The numbers in red in the table is abnormal may be caused by some reasons we did not find out.
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According to Figure 2, during fermentation, the content of acetic acid is slowly increased, verified that strains in our project can transfer carbon sources to other metabolites. Since acetic acid is a normal product of the metabolic cycle, we can detect an increase in the amount of acetic acid in the culture.
  
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The results above proved the function of the BBa_K3585003 in transforming galactose into acetic acid, through which fulfills the key stage of transforming galacose into butyrate.
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===Functional Parameters===
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<partinfo>BBa_K3585003 parameters</partinfo>
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Latest revision as of 13:58, 27 October 2020


J23100-butyrate

Continuous expression of butyrate synthesis pathway enzymes through a constitutive promoter, so the strain can degrade galactose and synthesize butyrate.

T--Shanghai HS United-BBa K3585003 fig 00.png



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1240
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 6272
    Illegal BglII site found at 7408
    Illegal BglII site found at 7549
    Illegal XhoI site found at 1245
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Contribution

BBa_K3585003 is a composite part consisting of butyrate synthesis pathway enzymes under constitutive promoter (basic parts BBa_J23100). The butyrate biosynthesis gene cluster can degrade galactose and synthesize butyrate.


Engineering success

The BBa_K3585003 was designed to transform galactose to butyrate, that is beneficial for galactosemia patients. In this process, the transformation of galactose to acetic acid is a crucial stage.

Following are the results of our functional tests of BBa_K3585003 in transforming galactose to acetic acid.

figure 1
                                            Figure 1 Chromatographic Peak of Acetic Acid
figure 2
                                     Figure 2 the Concentration of Acetic Acid while Fermentation

Each of the strains we have a repeat. Gal represents that we use galactose as the carbon source while fermentation; GG represents that we use both glucose and galactose as the carbon source while fermentation; 1917 represents wild type strains; pButy represents that the host strain contained pMTL83151-J23100-butyrate plasmids and expressed butyrate synthesis gene cluster; placY represents that the host strain contained p15A-J23200-lacY plasmids and expressed lacY gene; 1 and 2 represent for sample repeats. The numbers in red in the table is abnormal may be caused by some reasons we did not find out.

According to Figure 2, during fermentation, the content of acetic acid is slowly increased, verified that strains in our project can transfer carbon sources to other metabolites. Since acetic acid is a normal product of the metabolic cycle, we can detect an increase in the amount of acetic acid in the culture.

The results above proved the function of the BBa_K3585003 in transforming galactose into acetic acid, through which fulfills the key stage of transforming galacose into butyrate.