Difference between revisions of "Part:BBa K5416030"

 
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<partinfo>BBa_K5416030 short</partinfo>
 
<partinfo>BBa_K5416030 short</partinfo>
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<img src="https://static.igem.wiki/teams/5416/logo-name-t.png"  alt="Description of image" width="200" />
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<figcaption><strong>Imperial-College 2024</strong></figcaption>
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This part is designed by Team Imperial_College in iGEM 2024. Aiming to form an artificial organelle which serves as an enclosed chamber for natrual rubber production.
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This composite part encodes a polyhydroxyalkanoate(PHA) phasin protein PhaP ([[Part:BBa_K208001|BBa_K208001]]) and a fusion protein of PHA synthase PhaC from C. necators fused with rubber synthase HRT2trunc ([[Part:BBa_K5416000|BBa_K5416000]]). The CDS were placed in between a t7-lacO promoter ([[Part:BBa_K1624002|BBa_K1624002]]) and a t7 terminator ([[Part:BBa_K731721|BBa_K731721]]) for IPTG inducible expression. This part is designed to enable the formation of rubber particles as an artificial organelle through a mechanism inspired by the formation of PHA granules.
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This composite part, with two CDS [[Part:BBa_K208001|BBa_K208001]] and [[Part:BBa_K5416031|BBa_K5416031]] placed downstream of a pT7-LacO promoter [[Part:BBa_K2406020|BBa_K2406020]], is designed to produce an artificial organelle of rubber particle inspired by the formation of polyhydroxyalkanoate (PHA) carboxysomes.
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=Background=
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  <img src="https://static.igem.wiki/teams/5416/parts/phap-phac-hrt2trunc/fig1-background.png" width="400px">
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  <p><small>Fig 1: How PhaC makes PHA granules[1][2]</small></p>
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Polyhydroxyalkanoate, is a hydrophobic polymer made in many prokaryotes, and recently becoming a famous material as it can be used as a bio-degradable plastic. Whilst the polymer is very hydrophobic in nature, it can be readily produced in large quantities in prokaryotic system, comparing to rubber. Insights into the structure of the enzyme producing the PHA, the PHA synthase (PhaC), reveals the existence of a hydrophobic domain close to the active site. This domain is shown to be involved in the granule formation of the PHA and with absence of this domain, little PHA is made in the cell. This appease to be similar to the issue observed with bioproduction of natural rubber, where losing anchorage of the molecule results in significant reduction of the total amount of rubber produced.
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  <img src="https://static.igem.wiki/teams/5416/parts/phap-phac-hrt2trunc/fig2-illustration.png" width="400px">
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  <p><small>Fig 2: An illustration of how this part is designed to work</small></p>
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Inspired by the structure of PhaC, we questioned if the mechanism could be applied to the formation of rubber granules. Where the HRT2trunc (BBa_K5416000) is redesigned to be attached to the N terminus hydrophobic PHA binding domain of PhaC1. This would hypothetically capture the newly formed rubber chain and phase-separate the rubber granule with the rest of the cell. By introducing the other PHA-associated protein (PhaP), this structure can be then stabilized and become an ideal environment for rubber synthesis.
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=Design=
  
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===Usage and Biology===
 
  
 
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Revision as of 10:20, 30 September 2024


PhaP PhaC-HRT2trunc Description of image

Imperial-College 2024
This part is designed by Team Imperial_College in iGEM 2024. Aiming to form an artificial organelle which serves as an enclosed chamber for natrual rubber production.

This composite part, with two CDS BBa_K208001 and BBa_K5416031 placed downstream of a pT7-LacO promoter BBa_K2406020, is designed to produce an artificial organelle of rubber particle inspired by the formation of polyhydroxyalkanoate (PHA) carboxysomes.

Background

Fig 1: How PhaC makes PHA granules[1][2]

Polyhydroxyalkanoate, is a hydrophobic polymer made in many prokaryotes, and recently becoming a famous material as it can be used as a bio-degradable plastic. Whilst the polymer is very hydrophobic in nature, it can be readily produced in large quantities in prokaryotic system, comparing to rubber. Insights into the structure of the enzyme producing the PHA, the PHA synthase (PhaC), reveals the existence of a hydrophobic domain close to the active site. This domain is shown to be involved in the granule formation of the PHA and with absence of this domain, little PHA is made in the cell. This appease to be similar to the issue observed with bioproduction of natural rubber, where losing anchorage of the molecule results in significant reduction of the total amount of rubber produced.

Fig 2: An illustration of how this part is designed to work

Inspired by the structure of PhaC, we questioned if the mechanism could be applied to the formation of rubber granules. Where the HRT2trunc (BBa_K5416000) is redesigned to be attached to the N terminus hydrophobic PHA binding domain of PhaC1. This would hypothetically capture the newly formed rubber chain and phase-separate the rubber granule with the rest of the cell. By introducing the other PHA-associated protein (PhaP), this structure can be then stabilized and become an ideal environment for rubber synthesis.

Design

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]