Difference between revisions of "Part:BBa K4613002"

 
Line 5: Line 5:
 
ADH3 is an amidohydrolase derived from Stenotrophomonas acidaminiphila and forms an octamer in solution.
 
ADH3 is an amidohydrolase derived from Stenotrophomonas acidaminiphila and forms an octamer in solution.
 
ADH3 was reported to exhibit 57- to 35,000-fold higher activity than other enzymes and is the most efficient OTA-detoxifying enzyme reported thus far and can hydrolyze OTA to nontoxic ochratoxin α (OTα) and L-β-phenylalanine (Phe). Moreover, soluble protein expression of ADH3 in Escherichia coli has been realized.
 
ADH3 was reported to exhibit 57- to 35,000-fold higher activity than other enzymes and is the most efficient OTA-detoxifying enzyme reported thus far and can hydrolyze OTA to nontoxic ochratoxin α (OTα) and L-β-phenylalanine (Phe). Moreover, soluble protein expression of ADH3 in Escherichia coli has been realized.
 +
 +
 +
 +
<p style="text-align: center!important;"><b>Fig.1 SDS-PAGE analysis of purified enzymes. M, standard protein markers; 1, purified E. coli–expressed ADH3.</b></p>
 +
 +
 +
<html>
 +
<center><img src="https://static.igem.wiki/teams/4613/wiki/parts/parts/hplc-analysis-of-the-ota-adh3.png"with="1000" height="" width="500" height=""/></center>
 +
</html>
 +
 +
 +
<p style="text-align: center!important;"><b>Fig.2 HPLC analysis of the reaction mixtures catalyzed by WT and variant S88.</b></p>
 +
 +
 +
Because of its high efficiency and soluble expression in <em> Escherichia coli </em>, we used the variant S88E of ADH3 engineered by <em> Xiong L et al. </em>(2023) in our project to degrade OTA.
 +
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 18:04, 8 October 2023


ADH3

ADH3 is an amidohydrolase derived from Stenotrophomonas acidaminiphila and forms an octamer in solution. ADH3 was reported to exhibit 57- to 35,000-fold higher activity than other enzymes and is the most efficient OTA-detoxifying enzyme reported thus far and can hydrolyze OTA to nontoxic ochratoxin α (OTα) and L-β-phenylalanine (Phe). Moreover, soluble protein expression of ADH3 in Escherichia coli has been realized.


Fig.1 SDS-PAGE analysis of purified enzymes. M, standard protein markers; 1, purified E. coli–expressed ADH3.



Fig.2 HPLC analysis of the reaction mixtures catalyzed by WT and variant S88.


Because of its high efficiency and soluble expression in Escherichia coli , we used the variant S88E of ADH3 engineered by Xiong L et al. (2023) in our project to degrade OTA.


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 733
    Illegal AgeI site found at 421
    Illegal AgeI site found at 583
  • 1000
    COMPATIBLE WITH RFC[1000]