Difference between revisions of "Part:BBa K2617000"

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(→‎Enzyme activity)
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This halohydrin dehalogenase gene codes for a protein of 726 amino acids with a molecular mass of 82.3 kDa.
 
This halohydrin dehalogenase gene codes for a protein of 726 amino acids with a molecular mass of 82.3 kDa.
  
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===Enzyme activity ===
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===Enzyme activity of xylanase===
  
  
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[[File:T--UESTC-China-xylanase activity.png|500px|thumb|center|'''Fig. 2''' Xylanase Activity on Xylan. Data were measured inphosphate buffer solution at pH 6.0 and 40℃.]]
 
[[File:T--UESTC-China-xylanase activity.png|500px|thumb|center|'''Fig. 2''' Xylanase Activity on Xylan. Data were measured inphosphate buffer solution at pH 6.0 and 40℃.]]
  
 +
===Enzyme activity of xylanase===
 +
Ferulic acid esterase can decompose ferulic acid p-nitrophenol ester to produce p-nitrophenol and ferulic acid.
 +
The strain BL21 (DE3) transformed with piGEM2018-Module001 was cultivated overnight and centrifuged to obtain the supernatant. The remaining bacteria were broken and broken products were obtained. DSMO solution of ferulic acid p-nitrophenol ester was added to phosphate buffer solution of 630 L pH=6.4 with a concentration of 400 L of 10 mmol/L. Heat preservation 5min at 40℃ and add 0.2ml sample solution. The initial OD values were measured before incubation. Gently mix and incubate 4h at 40 C, determine the final OD value and compare the difference.
 
[[File:T--UESTC-China--Enzyme Activity of Ferulic acid esterase.png|500px|thumb|center|'''Fig. 3''' Enzyme Activity of Ferulic acid esterase. Data were measured in phosphate buffer solution at pH 6.4 and 40℃.]]
 
[[File:T--UESTC-China--Enzyme Activity of Ferulic acid esterase.png|500px|thumb|center|'''Fig. 3''' Enzyme Activity of Ferulic acid esterase. Data were measured in phosphate buffer solution at pH 6.4 and 40℃.]]
  

Revision as of 17:43, 10 October 2018


Xyn10D-Fae1A：Bifunctional xylanase/ferulic acid esterase

Xyn1D-fae1A cuts off the crosslinking between polysaccharides and lignin in cell wall and degrade xylose hemicellulose. This part is responsible for pretreatment of straw.

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 534
    Illegal AgeI site found at 381
    Illegal AgeI site found at 628
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1236

Characterization

Molecular weight

This halohydrin dehalogenase gene codes for a protein of 726 amino acids with a molecular mass of 82.3 kDa.

Enzyme activity of xylanase

Fig. 1 Standard Curve of Xylose. Data were measured in phosphate buffer solution at pH 6.0 and 40℃.
Fig. 2 Xylanase Activity on Xylan. Data were measured inphosphate buffer solution at pH 6.0 and 40℃.

Enzyme activity of xylanase

Ferulic acid esterase can decompose ferulic acid p-nitrophenol ester to produce p-nitrophenol and ferulic acid. The strain BL21 (DE3) transformed with piGEM2018-Module001 was cultivated overnight and centrifuged to obtain the supernatant. The remaining bacteria were broken and broken products were obtained. DSMO solution of ferulic acid p-nitrophenol ester was added to phosphate buffer solution of 630 L pH=6.4 with a concentration of 400 L of 10 mmol/L. Heat preservation 5min at 40℃ and add 0.2ml sample solution. The initial OD values were measured before incubation. Gently mix and incubate 4h at 40 C, determine the final OD value and compare the difference.

Fig. 3 Enzyme Activity of Ferulic acid esterase. Data were measured in phosphate buffer solution at pH 6.4 and 40℃.