Difference between revisions of "Part:BBa K4949002"
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+ | '''Figure 1.''' Michaelis-Menten Esterase Activity Assay of <i>Thermoanaerobacter thermohydrosulfuricus</i> 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. | ||
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+ | <html><img src="https://static.igem.wiki/teams/4949/wiki/registry/ttl-npo-mmeaa-graphs.png" width="950" height="700"></html> | ||
+ | '''Figure 2.''' Michaelis-Menten Esterase Activity Assay of <i>Thermoanaerobacter thermohydrosulfuricus</i> 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 ℃. |
Revision as of 02:15, 12 October 2023
MGS0156
MGS0156 is a serine dependent ⍺/β hydrolase originally identified from an environmental metagenomic analysis study to look for enzymes to degrade PLA
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 380
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 982
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 82
Illegal NgoMIV site found at 253 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 766
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 ℃.