Part:BBa_K4275007
FAST-PETase
FAST(Functional, Active, Stable, Tolerant)-PETase, is a thermostable, high-catalytic efficiency variant of the wild-type PET hydrolase from Ideonella sakaiensis. This variant contains five amino acid mutations (N233K/R224Q/S121E/D186H/R280A), designed via machine learning based on the original article[1]. The enzyme functions in an optimal temperature range of around 50℃, much lower than other artificially designed PETase variants (60-70℃). FAST-PETase catalyzes the cleavage of ester bonds between PET monomers, generating MHET(Monohydroxyethyl terephthalate) as the main product and a trace amount of BHET(bis-hydroxyethyl terephthalate) as the by-product. The enzyme FAST-PETase works in synergy with MHETase, the second enzyme in the two-enzyme PET degradation system found in Ideonella sakaiensis - a PET-degrading and assimilating strain of bacteria isolated from a water treatment plant.
Figure 1 The 3D structure of the protein predicted by Alphafold2.
Usage and Biology
The three-amino acid catalytic triad Ser131, His242, and Asp177 constitute the active site of the PET hydrolase, which catalyzes the reaction (ethylene terephthalate)n + H2O → (ethylene terephthalate)n-1 + MHET or BHET. The designation of the five optimal point mutations (N233K, R224Q, S121E, D186H, and R280A) is implemented via a convolutional neural network (CNN) called MutCompute[1] based on the original article. The algorithm learns the local chemical microenvironments of amino acids based on training over 19,000 sequence-balanced protein structures from the Protein Data Bank (PDB).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
1. Lu, Hongyuan et al. "Machine Learning-Aided Engineering Of Hydrolases For PET Depolymerization". Nature, vol 604, no. 7907, 2022, pp. 662-667. Springer Science And Business Media LLC, https://doi.org/10.1038/s41586-022-04599-z.
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