Difference between revisions of "Part:BBa K1360001"
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This part is the coding sequence for a thermostable alpha-L-arabinofuranosidase, which specifically releases arabinofuranoside branch of arabinoxylan from xylan. | This part is the coding sequence for a thermostable alpha-L-arabinofuranosidase, which specifically releases arabinofuranoside branch of arabinoxylan from xylan. | ||
| − | + | [[File:ArfB_decolorfour.png|thumb|]] | |
===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_Absorbance.png|center|frame||We also collect the absorbance data of the DNS. In this DNS method, OD540 corresponds to reductive saccharide concentration, but due to equipment limitation, we used OD570 for plate reader data.]] | ||
| + | ===Extra=== | ||
| + | [[File:Decolorfour.png|center|frame|In 2014, team Tongji's project Decolor Four.]] | ||
| + | [[File:Principle_Bleaching.gif|center|frame||The reason why these three hemicellulase can reduce 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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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Latest revision as of 04:38, 2 November 2014
alpha-L-arabinofuranosidase ArfB
This part is the coding sequence for a thermostable alpha-L-arabinofuranosidase, which specifically releases arabinofuranoside branch of arabinoxylan from xylan.
Usage and Biology
The ArfB 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 = 4.0, temperature = 71℃.
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.
We also collect the absorbance data of the DNS. In this DNS method, OD540 corresponds to reductive saccharide concentration, but due to equipment limitation, we used OD570 for plate reader data.
Extra
In 2014, team Tongji's project Decolor Four.
The reason why these three hemicellulase can reduce 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]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 679
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1156
Illegal BsaI.rc site found at 1282
Illegal BsaI.rc site found at 1372