Difference between revisions of "Part:BBa K1360003"
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<partinfo>BBa_K1360003 short</partinfo> | <partinfo>BBa_K1360003 short</partinfo> | ||
− | + | This part is the coding sequence for a thermostable mannosidase, which specifically degrades mannan. | |
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===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 optimum working condition is pH = 5.0, temperature = 75.4℃. | ||
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+ | ===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.]] | ||
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+ | [[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.]] | ||
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+ | [[File:DNS_Plate_Area.png|center|frame||We carried out temperature test, pH test and synergy test on one plate.]] | ||
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+ | [[File:DNS_Temperature.png|center|frame||The result of temperature test.]] | ||
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+ | [[File:DNS_pH.png|center|frame||The result of pH test.]] | ||
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+ | [[File:DNS_Synergy1.png|center|frame||As for synergy test, we add three enzymes at different ratio.]] | ||
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+ | [[File:DNS_Synergy2.png|center|frame||Firstly, we recorded DNS color when there is only one enzyme.]] | ||
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+ | [[File:DNS_Synergy3.png|center|frame||And then, we supposed they have no synergistic effect, and simulate the RGB color.]] | ||
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+ | [[File:DNS_Synergy4.png|center|frame||Lastly, we compared the real DNS color with the non-synergy supposed DNS color.]] | ||
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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.]] | ||
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Revision as of 18:48, 1 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
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