Difference between revisions of "Part:BBa K3241040"
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<partinfo>BBa_K3241040 short</partinfo>
 
<partinfo>BBa_K3241040 short</partinfo>
  
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    This part is a new part, which consists of BBa_K3241000 BBa_K3241022 BBa_K3241112 BBa_K2800032 BBa_K2800013. It is used to construct the PHB biosynthetic pathway in Zymomonas mobilis .
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This part is a new part, which consists of BBa_K3241000 BBa_K3241022 BBa_K3241112 BBa_K2800032 BBa_K2800013. It is used to construct the PHB biosynthetic pathway in Zymomonas mobilis .
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    Z. mobilis mainly metabolizes glucose by Entner-Doudoroff pathway to produce ethanol. Here，we introduced phbA, phbB, and phbC genes from Ralstonia eutropha to modify its metabolic pathway. The expression of these genes make acetyl-coA flows into PHB synthesis.
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    [[File:T--HUBU_WUHAN--HeterogenousPHB_biosynthesis_pathway.jpeg|300px|left|Fig.1 Heterogenous PHB biosynthesis pathway]]
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    [[File:T--HUBU_WUHAN--enzymes.jpg|300px|right|Fig.2 PHB synthesizes key enzymes]
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Part one:Basic Fermentation
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    In our basic fermentation Ptet was used to explore the growth of recombinant strains under the induction of different concentrations of tetracycline.For more information, please refer to BBa_K3241029.
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    [[File:T--HUBU_WUHAN--PHB_pathway_genes.png|400px|left|Fig.1 PHB pathway genes.]]
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==Design Notes==
  
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<i>Zymomonas mobilis </i>is the model strain of ethanol production. It's mainly metabolize glucose, fructose and sucrose through ED(entner-doudoroff) pathway to produce ethanol. Here，we introduced phbA, phbB, and phbC genes from Ralstonia eutropha to modify its metabolic pathway.When Z.mobilis disassemble sugar molecules to produce pyruvate,  One part of the Metabolic flux produces ethanol under the catalysis of pyruvate decarboxylase and ethanol dehydrogenase, and in another part  pyruvate oxidation decarboxylation to acetyl CoA.Acetyl coenzyme A translates into PHB under the catalysis of β-ketothiolase, acetoacetyl-CoA Reductase and PHB polymerase.
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[[File:T--HUBU_WUHAN--HeterogenousPHB_biosynthesis_pathway.jpeg|300px|thumb|center|Fig.1 Heterogenous PHB biosynthesis pathway]]
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[[File:T--HUBU_WUHAN--enzymes.jpg|300px|center|thumb|Fig.2 PHB synthesizes key enzymes]]
  
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===Part one: basic fermentation===
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We used the promter Ptet to explore the growth of recombinant strains under the induction of different concentrations of tetracycline.(the original can refer to BBa_K3241029)
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[[File:T--HUBU WUHAN--Original design.png|300px|center|thumb| Fig.3 Original design.]]
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[[File:T--HUBU WUHAN--Growth Curve aasd.jpg|500px|left|thumb| Fig.4 Growth Curve.]]
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[[File:T--HUBU WUHAN--PHB Profuction.jpg|500px|right|thumb| Fig.5 PHB Production.]]
  
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==Sequence and Features==
  
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<span class='h3bb'>Sequence and Features</span>
  
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
 
 
<partinfo>BBa_K3241040 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3241040 SequenceAndFeatures</partinfo>
  

Revision as of 22:01, 16 October 2019


Pgap-PhbC-PhbA-PhbB

This part is a new part, which consists of BBa_K3241000 BBa_K3241022 BBa_K3241112 BBa_K2800032 BBa_K2800013. It is used to construct the PHB biosynthetic pathway in Zymomonas mobilis .

Design Notes

Zymomonas mobilis is the model strain of ethanol production. It's mainly metabolize glucose, fructose and sucrose through ED(entner-doudoroff) pathway to produce ethanol. Here，we introduced phbA, phbB, and phbC genes from Ralstonia eutropha to modify its metabolic pathway.When Z.mobilis disassemble sugar molecules to produce pyruvate, One part of the Metabolic flux produces ethanol under the catalysis of pyruvate decarboxylase and ethanol dehydrogenase, and in another part pyruvate oxidation decarboxylation to acetyl CoA.Acetyl coenzyme A translates into PHB under the catalysis of β-ketothiolase, acetoacetyl-CoA Reductase and PHB polymerase.

Fig.1 Heterogenous PHB biosynthesis pathway
Fig.2 PHB synthesizes key enzymes

Part one: basic fermentation

We used the promter Ptet to explore the growth of recombinant strains under the induction of different concentrations of tetracycline.(the original can refer to BBa_K3241029)

Fig.3 Original design.
Fig.4 Growth Curve.
Fig.5 PHB Production.

Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 465
    Illegal NheI site found at 1341
    Illegal NheI site found at 3257
    Illegal NotI site found at 2
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 495
    Illegal NgoMIV site found at 2356
    Illegal NgoMIV site found at 2441
    Illegal NgoMIV site found at 2846
    Illegal NgoMIV site found at 2975
    Illegal AgeI site found at 600
    Illegal AgeI site found at 3379
    Illegal AgeI site found at 4051
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 421
    Illegal BsaI site found at 1142
    Illegal BsaI site found at 2272