Part:BBa_K4701301
lamB-MHETase
Introduction
Enzymatic biodegradation of polyethylene terephthalate (PET) as a tool to combat accumulating plastic has been an intense area of research during recent years [1]. One promising pathway for PET degradation was identified in a novel strain I. sakaiensis, isolated from a Japanese landfill back in 2016 [2]. The primary enzyme IsPETase breaks the PET chain (see: BBa_K4701300), but in order to degrade the intermediate product mono(2-hydroxyethyl) terephthalate (MHET) all the way into the constituent monomers of PET terephthalic acid (TPA), and ethylene glycol (EG), a secondary enzyme called MHETase (EC 3.1.1.102) is used by I. sakaiensis.
As with IsPETase, MHETase is also secreted by I. sakaiensis. This part is an attempt to mimic this behavior with E. coli as the host organism. MHETase has previously been secreted from E. coli with various signal sequences, with one recent study finding lamB to perform the best [3]. We hope to provide a protein coding sequences that when clone into E. coli, allows for efficient secretion of a MHET hydrolase that can subsequently be purified.
See the design page for the detailed design process.Cloning strategy
References
[1] Qi, X., et al. (2022). Current advances in the biodegradation and bioconversion of polyethylene terephthalate. Microorganisms, 10(1). https://doi.org/10.3390/microorganisms10010039
[2] Yoshida, S., et al (2016). A bacterium that degrades and assimilates poly(ethylene terephthalate). Science. 351(6278), 1196-9. https://doi.org/10.1126/science.aad6359
[3] Sagong, H., et al (2022). Decomposition of the PET Film by MHETase Using Exo-PETase Function. ACS Catalysis. 10(8), 4805–4812. https://doi.org/10.1021/acscatal.9b05604
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]
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