Any feedback?
Literature summary extracted from
- SpyTag/SpyCatcher cyclization enhances the thermostability of firefly luciferase (2016), PLoS ONE, 11, e0162318 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.13.12.7 | recombinant coexpression of the luciferase with fused cyclization proteins SpyCatcher at the N-terminus and SpyTag at the C-terminus (pyCatcher-LUC-SpyTag) in Escherichia coli strain BL21(DE3) | Photinus pyralis |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.13.12.7 | additional information | firefly luciferase from Photinus pyralis is cyclized in vivo by fusing SpyCatcher at the N-terminus and SpyTag at the C-terminus. SpyTag can spontaneously form a covalent isopeptide bond with its protein partner Spy-Catcher. Circular LUC is more thermostable at 25-55°C and alkali-tolerant than the wild-type, without compromising the specific activity. Preparation of an N-terminally and C-terminally shortened form of the SpyCatcher protein and enzyme cyclization using this truncated form leads to even more thermostability of the construct than the full-length SpyCatcher form, method optimization | Photinus pyralis |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.13.12.7 | D-firefly luciferin + O2 + ATP | Photinus pyralis | - |
firefly oxyluciferin + CO2 + AMP + diphosphate + hv | - |
? |  |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.13.12.7 | Photinus pyralis | P08659 | - |
- |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.13.12.7 | D-firefly luciferin + O2 + ATP | - |
Photinus pyralis | firefly oxyluciferin + CO2 + AMP + diphosphate + hv | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.13.12.7 | ? | x * 61000, recombinant wild-type enzyme, SDS-PAGE, x * 80000, recombinant cyclized enzyme SpyCatcher-LUC-SpyTag, SDS-PAGE | Photinus pyralis |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.13.12.7 | firefly luciferase | - |
Photinus pyralis |
| 1.13.12.7 | Luc | - |
Photinus pyralis |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.13.12.7 | 25 | - |
wild-type enzyme | Photinus pyralis |
| 1.13.12.7 | 35 | - |
recombinant circular enzyme | Photinus pyralis |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.13.12.7 | 10 | 60 | temperature activity range, recombinant wild-type enzyme and recombinant circular enzyme | Photinus pyralis |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.13.12.7 | 16.7 | - |
Tm value of recombinant wild-type LUC | Photinus pyralis |
| 1.13.12.7 | 37.4 | - |
T50 for recombinant wild-type LUC | Photinus pyralis |
| 1.13.12.7 | 40.3 | - |
T1/2 value of recombinant cyclized LUC | Photinus pyralis |
| 1.13.12.7 | 47.5 | - |
T50 for recombinant cyclized LUC | Photinus pyralis |
| 1.13.12.7 | 49.2 | - |
T50 value of recombinant cyclized LUC with truncated Spy-Catcher cyclization protein | Photinus pyralis |
| 1.13.12.7 | 51.4 | - |
T1/2 value of recombinant cyclized LUC with truncated Spy-Catcher cyclization protein | Photinus pyralis |
| 1.13.12.7 | 53.5 | - |
Tm value of recombinant wild-type LUC | Photinus pyralis |
| 1.13.12.7 | 70.2 | - |
Tm value of recombinant cyclized LUC | Photinus pyralis |
| 1.13.12.7 | 73.1 | - |
Tm value of recombinant cyclized LUC with truncated Spy-Catcher cyclization protein | Photinus pyralis |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.13.12.7 | 8 | - |
wild-type enzyme | Photinus pyralis |
| 1.13.12.7 | 9 | - |
recombinant circular enzyme | Photinus pyralis |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.13.12.7 | 2.5 | 10.5 | pH activity range, recombinant circular enzyme | Photinus pyralis |
| 1.13.12.7 | 4 | 10 | pH activity range, recombinant wild-type enzyme | Photinus pyralis |
pH Stability
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.13.12.7 | 8 | 10 | recombinant circular LUC is stable at a pH range of 8.0-10.0, maintaining nearly 100% activity after incubation in various pH buffers for 30 min. Recombinant wild-type LUC maintains only 80% of its original activity after incubating at pH 10.0 for 30 min | Photinus pyralis |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.13.12.7 | ATP | - |
Photinus pyralis | |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
