Difference between revisions of "Part:BBa K1360003"
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This part is the coding sequence for a thermostable mannosidase, which specifically degrades mannan. | This part is the coding sequence for a thermostable mannosidase, which specifically degrades mannan. | ||
| − | + | [[File:XynB_decolorfour.png|center|frame||]] | |
===Usage and Biology=== | ===Usage and Biology=== | ||
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[[File:DNS_Synergy5.png|center|frame||As you can see, the real DNS color is darker, so we think they can work in synergy.]] | [[File:DNS_Synergy5.png|center|frame||As you can see, the real DNS color is darker, so we think they can work in synergy.]] | ||
| + | [[File:DNS_Plate_Absorbance.png|center|frame||We also collect the absorbance data of the DNS. ]] | ||
| + | ===Extra=== | ||
| + | [[File:Decolorfour.png|center|frame||In 2014, team Tongji's project Decolor Four. Here is our wiki: 2014.igem.org/Team:Tongji]] | ||
| + | |||
| + | [[File:Principle_Bleaching.gif|center|frame||The reason these three hemicellulase can reduced chlorine consumption is because they break down hemicellulose which connects lignin and cellulose. In this way, bleaching detergent can "get into" the pulp easier, and less chlorine can make the same effect. This benefits environmental protection.]] | ||
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Revision as of 04:24, 2 November 2014
endo-1,4-beta-mannosidase ManA1
This part is the coding sequence for a thermostable mannosidase, which specifically degrades mannan.
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 optimum working condition is pH = 5.0, temperature = 75.4℃.
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
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.
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.
We carried out temperature test, pH test and synergy test on one plate.
The result of temperature test.
The result of pH test.
As for synergy test, we add three enzymes at different ratio.
Firstly, we recorded DNS color when there is only one enzyme.
And then, we supposed they have no synergistic effect, and simulate the RGB color.
Lastly, we compared the real DNS color with the non-synergy supposed DNS color.
As you can see, the real DNS color is darker, so we think they can work in synergy.
File:DNS Plate Absorbance.png
We also collect the absorbance data of the DNS.
Extra
In 2014, team Tongji's project Decolor Four. Here is our wiki: 2014.igem.org/Team:Tongji
The reason these three hemicellulase can reduced chlorine consumption is because they break down hemicellulose which connects lignin and cellulose. In this way, bleaching detergent can "get into" the pulp easier, and less chlorine can make the same effect. This benefits environmental protection.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 552
Illegal BamHI site found at 1170 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 544
Illegal SapI.rc site found at 459