Difference between revisions of "Part:BBa K1201000"

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Chitinase gene; ChiA. Is a processive enzyme which degrades chitin from the reducing end. Suitable for projects which require an enzyme that degrades chitin.
 
Chitinase gene; ChiA. Is a processive enzyme which degrades chitin from the reducing end. Suitable for projects which require an enzyme that degrades chitin.
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Characterization
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We add 1 mL 200 m g / L of substrate to a clean test tube, 3mL 0.05 mol / L phosphate buffer , 3 min 50 ° C water bath temperature protection Min, and add 1 mL enzyme solution, mix and heat in 50 degrees water 15 min. After that, we used boiling water to terminate the enzymatic reaction, and add water to a volume of 10 mL. Evenly, the solution was centrifuged at 3000r /min for 10 min. Measure the OD value of the upper clear.
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Definition of enzyme unit: The amount of enzyme required to produce 1 gram of p-nitroaniline per hour under the above reaction conditions is defined as 1 enzyme activity  ( U /m L ).
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The calculation formula is: (A-A0)*Enzyme dilution factor/kt.                                                                                                                               
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In the formula:
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A: The absorbed value of the enzymatic hydrolyzate sample after constant volume A is fixed
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A0: The absorbance of the post-enzymatic solution blank
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T : Enzymatic reaction time, unit: h
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k, linear coefficient, 0.0648
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 22:33, 17 October 2018

Derived from serratia marcescens

Chitinase gene; ChiA. Is a processive enzyme which degrades chitin from the reducing end. Suitable for projects which require an enzyme that degrades chitin.

Characterization We add 1 mL 200 m g / L of substrate to a clean test tube, 3mL 0.05 mol / L phosphate buffer , 3 min 50 ° C water bath temperature protection Min, and add 1 mL enzyme solution, mix and heat in 50 degrees water 15 min. After that, we used boiling water to terminate the enzymatic reaction, and add water to a volume of 10 mL. Evenly, the solution was centrifuged at 3000r /min for 10 min. Measure the OD value of the upper clear. Definition of enzyme unit: The amount of enzyme required to produce 1 gram of p-nitroaniline per hour under the above reaction conditions is defined as 1 enzyme activity ( U /m L ).

The calculation formula is: (A-A0)*Enzyme dilution factor/kt. In the formula: A: The absorbed value of the enzymatic hydrolyzate sample after constant volume A is fixed

A0: The absorbance of the post-enzymatic solution blank

T : Enzymatic reaction time, unit: h

k, linear coefficient, 0.0648

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1227
    Illegal NgoMIV site found at 1678
    Illegal AgeI site found at 274
    Illegal AgeI site found at 1381
  • 1000
    COMPATIBLE WITH RFC[1000]