Difference between revisions of "Part:BBa K5023003"
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FAST-PETase is a mutant variant derived from the PETase enzyme, optimized with specific codon changes to enhance its expression efficiency in Chlamydomonas reinhardtii. This enzyme showcased superior PET-hydrolytic activity in comparison to both the wild-type and other engineered alternatives between 30 and 50°C across various pH levels. At 50°C, FAST-PETase exhibited the most significant degradation among all mutants tested, releasing 33.8 mM of PET monomers, which is the combined amount of terephthalic acid (TPA) and mono-(2-hydroxyethyl) terephthalate (MHET). Its activity against post-consumer PET (pc-PET) surpassed other enzymes like WT PETase, ThermoPETase, DuraPETase, LCC, and ICCM under identical conditions. Remarkably, FAST-PETase managed to degrade nearly all untreated post-consumer PET from 51 distinct thermoformed products within a week. Additionally, it proved its ability to depolymerize the untreated, amorphous sections of a commercial water bottle. A time-course analysis highlighted an almost linear PET degradation rate and a simultaneous rise in crystallinity over time. The introduction of FAST-PETase presents a promising approach to PET plastic degradation, especially considering its amplified activity and stability across diverse conditions. This enzyme holds significant potential in addressing the environmental issues caused by the accumulation of PET plastics. | FAST-PETase is a mutant variant derived from the PETase enzyme, optimized with specific codon changes to enhance its expression efficiency in Chlamydomonas reinhardtii. This enzyme showcased superior PET-hydrolytic activity in comparison to both the wild-type and other engineered alternatives between 30 and 50°C across various pH levels. At 50°C, FAST-PETase exhibited the most significant degradation among all mutants tested, releasing 33.8 mM of PET monomers, which is the combined amount of terephthalic acid (TPA) and mono-(2-hydroxyethyl) terephthalate (MHET). Its activity against post-consumer PET (pc-PET) surpassed other enzymes like WT PETase, ThermoPETase, DuraPETase, LCC, and ICCM under identical conditions. Remarkably, FAST-PETase managed to degrade nearly all untreated post-consumer PET from 51 distinct thermoformed products within a week. Additionally, it proved its ability to depolymerize the untreated, amorphous sections of a commercial water bottle. A time-course analysis highlighted an almost linear PET degradation rate and a simultaneous rise in crystallinity over time. The introduction of FAST-PETase presents a promising approach to PET plastic degradation, especially considering its amplified activity and stability across diverse conditions. This enzyme holds significant potential in addressing the environmental issues caused by the accumulation of PET plastics. | ||
| − | ===Reference | + | ===Reference=== |
Lu, H., Diaz, D.J., Czarnecki, N.J. et al. Machine learning-aided engineering of hydrolases for PET depolymerization. Nature 604, 662–667 (2022). https://doi.org/10.1038/s41586-022-04599-z | Lu, H., Diaz, D.J., Czarnecki, N.J. et al. Machine learning-aided engineering of hydrolases for PET depolymerization. Nature 604, 662–667 (2022). https://doi.org/10.1038/s41586-022-04599-z | ||
Latest revision as of 20:30, 8 October 2023
FAST-PETase
FAST-PETase is a mutant variant derived from the PETase enzyme. It contains five mutations compared to the wild-type PETase. These mutations include N233K, R224Q, and S121E from prediction, along with D186H and R280A from the scaffold. This enzyme demonstrated superior PET-hydrolytic activity relative to both wild-type and other engineered alternatives between 30 and 50°C across a range of pH levels. At 50°C, FAST-PETase showed the highest overall degradation of all mutants tested, releasing 33.8 mM of PET monomers (the sum of terephthalic acid (TPA) and mono-(2-hydroxyethyl) terephthalate (MHET)). The enzyme's activity against post-consumer PET (pc-PET) was substantially higher than that of other enzymes like WT PETase, ThermoPETase, DuraPETase, LCC, and ICCM under the same conditions. FAST-PETase was able to almost completely degrade untreated post-consumer PET from 51 different thermoformed products in just one week. The enzyme also demonstrated the capability to depolymerize untreated, amorphous portions of a commercial water bottle. A time-course analysis revealed an almost linear decay rate of PET degradation and a concomitant increase in crystallinity over time. The development of FAST-PETase offers a potential solution for the degradation of PET plastics, especially given its enhanced activity and stability across a range of conditions. This enzyme can play a pivotal role in addressing the environmental challenges posed by PET plastic accumulation. Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 846
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 846
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 351
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 846
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 846
Illegal NgoMIV site found at 155 - 1000COMPATIBLE WITH RFC[1000]
Improvement on BBa_K5023003 by UNILA-Latam 2023
We improved upon the FAST-PETase part (Part:BBa_K4390073) with the FAST-PETase part (Part:BBa_K5023003) optimized for Chlamydomonas reinhardtii.
FAST-PETase is a mutant variant derived from the PETase enzyme, optimized with specific codon changes to enhance its expression efficiency in Chlamydomonas reinhardtii. This enzyme showcased superior PET-hydrolytic activity in comparison to both the wild-type and other engineered alternatives between 30 and 50°C across various pH levels. At 50°C, FAST-PETase exhibited the most significant degradation among all mutants tested, releasing 33.8 mM of PET monomers, which is the combined amount of terephthalic acid (TPA) and mono-(2-hydroxyethyl) terephthalate (MHET). Its activity against post-consumer PET (pc-PET) surpassed other enzymes like WT PETase, ThermoPETase, DuraPETase, LCC, and ICCM under identical conditions. Remarkably, FAST-PETase managed to degrade nearly all untreated post-consumer PET from 51 distinct thermoformed products within a week. Additionally, it proved its ability to depolymerize the untreated, amorphous sections of a commercial water bottle. A time-course analysis highlighted an almost linear PET degradation rate and a simultaneous rise in crystallinity over time. The introduction of FAST-PETase presents a promising approach to PET plastic degradation, especially considering its amplified activity and stability across diverse conditions. This enzyme holds significant potential in addressing the environmental issues caused by the accumulation of PET plastics.
Reference
Lu, H., Diaz, D.J., Czarnecki, N.J. et al. Machine learning-aided engineering of hydrolases for PET depolymerization. Nature 604, 662–667 (2022). https://doi.org/10.1038/s41586-022-04599-z