Difference between revisions of "Part:BBa K5291048:Design"

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===Design Notes===
 
===Design Notes===
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In degradation module we constructed 4 main plasmids. Two PE degrading enzymes, CYPY96F-VHB and AlkB<sub>2</sub>-Rd45-AdhA, are designed to accelerate the degradation rate by improving the absorbing and hydroxylation of PE monomer. In order to express above two enzymes in our chassis， pAB1-CYPY96F-VHB and pAB1-AlkB<sub>2</sub>-Rd45-VHB were constructed. pAB1-pS-PEBP-PEase was constructed to depolymerize PE microplastics. In addition, pAB1-PEBP-GFP was constructed to verify the adsorption capacity of PEBP to PE by fluorescence microscopy. AlkB<sub>2</sub> is a significant component in degradation module.
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Alkanes are the most energy-rich form of carbon and are widely dispersed in the environment. Their transformation by microbes represents a key step in the global carbon cycle. Alkane monooxygenase (AlkB), a membrane-spanning metalloenzyme, converts straight chain alkanes to alcohols in the first step of the microbially-mediated degradation of alkanes, thereby playing a critical role in the global cycling of carbon and the bioremediation of oil. AlkB biodiversity is attributed to its ability to oxidize alkanes of various chain lengths, while individual AlkBs target a relatively narrow range. Mechanisms of substrate selectivity and catalytic activity remain elusive. Here we report the cryo-EM structure of AlkB, which provides a distinct architecture for membrane enzymes. Our structure and functional studies reveal an unexpected diiron center configuration and identify molecular determinants for substrate selectivity. These findings provide insight into the catalytic mechanism of AlkB and shed light on its function in alkane-degrading microorganisms.
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===Source===
 
===Source===
  
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===References===
 
===References===
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[1]Guo, X., et al. (2023). "Structure and mechanism of the alkane-oxidizing enzyme AlkB." Nat Commun 14(1): 2180.<br>
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[2]Yuan, M., et al. (2020). "Selective Electroenzymatic Oxyfunctionalization by Alkane Monooxygenase in a Biofuel Cell." Angewandte Chemie. 59(23): 8969-8973.

Latest revision as of 06:29, 2 October 2024


AlkB2


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 657
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Alkanes are the most energy-rich form of carbon and are widely dispersed in the environment. Their transformation by microbes represents a key step in the global carbon cycle. Alkane monooxygenase (AlkB), a membrane-spanning metalloenzyme, converts straight chain alkanes to alcohols in the first step of the microbially-mediated degradation of alkanes, thereby playing a critical role in the global cycling of carbon and the bioremediation of oil. AlkB biodiversity is attributed to its ability to oxidize alkanes of various chain lengths, while individual AlkBs target a relatively narrow range. Mechanisms of substrate selectivity and catalytic activity remain elusive. Here we report the cryo-EM structure of AlkB, which provides a distinct architecture for membrane enzymes. Our structure and functional studies reveal an unexpected diiron center configuration and identify molecular determinants for substrate selectivity. These findings provide insight into the catalytic mechanism of AlkB and shed light on its function in alkane-degrading microorganisms.

Source

Pseudomonas aeruginosa PAO1

References

[1]Guo, X., et al. (2023). "Structure and mechanism of the alkane-oxidizing enzyme AlkB." Nat Commun 14(1): 2180.
[2]Yuan, M., et al. (2020). "Selective Electroenzymatic Oxyfunctionalization by Alkane Monooxygenase in a Biofuel Cell." Angewandte Chemie. 59(23): 8969-8973.