Difference between revisions of "Part:BBa K2910000"

Revision as of 03:02, 22 October 2019

IsPETase-W159H-S238F with C-terminal Hexahistidine Tag

BBa_K2910000 encodes an alpha/beta-hydrolase that degrades poly(ethylene terephthalate) (PET) into mono(2-hydroxyethyl) terephthalic acid (MHET) with trace amounts of terephthalic acid (TPA), and bis(2-hydroxyethyl)-TPA (BHET). The enzyme originates from Ideonella sakaiensis, a microbe that was discovered to assimilate PET plastic in 2016 by Yoshida et al. This part has been codon-optimized for E. coli K-12 and contains a C-terminal hexahistidine tag for affinity protein purification. Additionally, the part contains two mutations that result in the nonsynonymous amino acid substitutions W159H and S238F relative to the wildtype sequence. These mutations enhance the catalytic activity of the enzyme by narrowing the active site, which makes PETase more "cutinase-like" according to X-ray crystallography structure analysis by Austin et al (2018).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NotI site found at 67
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 ===Usage and Biology===
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 <span class='h3bb'>Sequence and Features</span>
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