Coding
TTL

Part:BBa_K4949001

Designed by: UManitoba 2023   Group: iGEM23_UManitoba   (2023-10-09)
Revision as of 02:07, 12 October 2023 by Prefont5 (Talk | contribs)


Thermoanaerobacter thermohydrosulfuricus lipase (TTL)

Thermoanaerobacter thermohydrosulfuricus lipase (TTL) is a thermophilic lipase that has been studied in the potential degradation of polyethylene terephthalate (PET). This protein expresses well in E.coli and sufficient quantity can be purified by IMAC test is ability to cleave ester linkages.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Figure 1. Michaelis-Menten Esterase Activity Assay of Thermoanaerobacter thermohydrosulfuricus lipase with 4-nitrophenyl butyrate (pNOB). Assays were conducted with 1.5 µM of lipase in PBS with pNOB concentration ranging from 0.0365 - 0.11 µM. The rate of change of absorbance was monitored at 410 nm for 1 min. A) Wild-type TTL lipase chromogenic assay with initial rate as a function of substrate concentration at 25 B) Wild-type TTL lipase chromogenic assay with initial rate as a function of substrate concentration at 60 ℃ C) TTL lipase chromogenic assay with initial rate as a function of substrate concentration, with norleucine introduced by residue specifically by SPI at 25 ℃ D) TTL lipase chromogenic assay with initial rate as a function of substrate concentration, with norleucine introduced by residue.


Figure 2. Michaelis-Menten Esterase Activity Assay of Thermoanaerobacter thermohydrosulfuricus lipase with 4-nitrophenyl octanoate (NPO). Assays were conducted with 1.5 µM of lipase in PBS with pNOB concentration ranging from 0.0365-0.11 µM. The rate of change of absorbance was monitored at 410 nm for 1 min. A) Wild-type TTL lipase chromogenic assay with initial rate as a function of substrate concentration at 25 ℃ B) Wild-type TTL lipase chromogenic assay with initial rate as a function of substrate concentration at 60 ℃. C) TTL lipase chromogenic assay with initial rate as a function of substrate concentration, with norleucine introduced by residue specifically by SPI at 25℃. D) TTL lipase chromogenic assay with initial rate as a function of substrate concentration, with norleucine introduced by residue specifically by SPI at 60 ℃.

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Parameters
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