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. | ||
+ | |||
+ | 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 | ||
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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
- 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 1227
Illegal NgoMIV site found at 1678
Illegal AgeI site found at 274
Illegal AgeI site found at 1381 - 1000COMPATIBLE WITH RFC[1000]