Difference between revisions of "Part:BBa K1360002"
Yuiuiuiuiui (Talk | contribs) |
|||
Line 1: | Line 1: | ||
− | |||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1360002 short</partinfo> | <partinfo>BBa_K1360002 short</partinfo> | ||
− | + | This part is the coding sequence for a thermostable xylanase, which specifically degrades xylan. | |
+ | |||
− | |||
===Usage and Biology=== | ===Usage and Biology=== | ||
+ | The XynB enzyme produced by this part can be used in pulp bleaching. It presents high activity under a wide range of temperature (55℃-99℃) and pH conditions (3-10.5). The optimized working condition is pH = 6.0, temperature = 65.9℃. This enzyme is expressed at high level in E.coli BL21(DE3) strain after T7 promoter. | ||
+ | |||
+ | ===Design considerations=== | ||
+ | This part is only a coding sequence without expression regulators, and is compatible with RFC10 standards, and the codon is optimized for E.coli expression. | ||
+ | |||
+ | ===Lab Archives=== | ||
+ | [[File:DNS_Abstract.png|center|frame||We use DNS to test the enzymatic activity. DNS is a reagent detecting reductive saccharides. When mannan or xylan is degraded into mannose or xylose, DNS color get dark. When DNS color darker, the enzymatic activity higher.]] | ||
+ | |||
+ | [[File:DNS_Plate.png|center|frame||In 2014, team Tongji carried out DNS enzymatic experiment of xynB, arfB and manA1 on one plate, so that you can get conclusion in a very straightforward way.]] | ||
+ | |||
+ | [[File:DNS_Plate_Area.png|center|frame||We carried out temperature test, pH test and synergy test on one plate.]] | ||
+ | |||
+ | [[File:DNS_Temperature.png|center|frame||The result of temperature test.]] | ||
+ | |||
+ | [[File:DNS_pH.png|center|frame||The result of pH test.]] | ||
+ | |||
+ | [[File:DNS_Synergy1.png|center|frame||As for synergy test, we add three enzymes at different ratio.]] | ||
+ | |||
+ | [[File:DNS_Synergy2.png|center|frame||Firstly, we recorded DNS color when there is only one enzyme.]] | ||
+ | |||
+ | [[File:DNS_Synergy3.png|center|frame||And then, we supposed they have no synergistic effect, and simulate the RGB color.]] | ||
+ | |||
+ | [[File:DNS_Synergy4.png|center|frame||Lastly, we compared the real DNS color with the non-synergy supposed DNS color.]] | ||
+ | |||
+ | [[File:DNS_Synergy5.png|center|frame||As you can see, the real DNS color is darker, so we think they can work in synergy.]] | ||
+ | |||
<!-- --> | <!-- --> |
Revision as of 18:46, 1 November 2014
endo-1,4-beta-xylanase XynB
This part is the coding sequence for a thermostable xylanase, which specifically degrades xylan.
Usage and Biology
The XynB enzyme produced by this part can be used in pulp bleaching. It presents high activity under a wide range of temperature (55℃-99℃) and pH conditions (3-10.5). The optimized working condition is pH = 6.0, temperature = 65.9℃. This enzyme is expressed at high level in E.coli BL21(DE3) strain after T7 promoter.
Design considerations
This part is only a coding sequence without expression regulators, and is compatible with RFC10 standards, and the codon is optimized for E.coli expression.
Lab Archives
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 847
Illegal AgeI site found at 610 - 1000COMPATIBLE WITH RFC[1000]