Difference between revisions of "Part:BBa K4192021"
 
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It is one of the genes required for oxalic acid synthesis in Burkholderia glumae, which can act together with ObcB to catalyze oxaloacetic acid (OAA) and citric acid to form oxalic acid (OA). When there is only ObcB, it cannot form oxalic acid.
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It is one of the genes required for oxalic acid synthesis in <i>Burkholderia glumae</i>, which can act together with ObcB to catalyze oxaloacetic acid (OAA) and citric acid to form oxalic acid (OA). When there is only ObcB, it cannot form oxalic acid.
  
 
===Biology===
 
===Biology===

Latest revision as of 15:08, 12 October 2022


obcB, one of the genes required for oxalic acid synthesis

It is one of the genes required for oxalic acid synthesis in Burkholderia glumae, which can act together with ObcB to catalyze oxaloacetic acid (OAA) and citric acid to form oxalic acid (OA). When there is only ObcB, it cannot form oxalic acid.

Biology

CAU_China 2022 used this part in their oxalate secretion circuit and the expression product of obcB is one of the main enzyme in in the oxalate secretion circuit[1]. It can act together with ObcA to catalyze oxaloacetic acid (OAA) and citric acid to form oxalic acid (OA). When there is only ObcB, it cannot form oxalic acid[2].

CAU-obcAB-pic.jpeg

Fig.1 Effects of obcA and obcB, the shade of purple reflects the concentration of oxalic acid[1].


When producing oxalic acid, obcB turns C6-CoA adduct into CoA, acetoacetate,and oxalic acid


CAU-obcAB-work.jpeg

Fig.2 Mechanism of obcA and obcB[2].

Considering the importance of obcA and obcB occur togrther in the biosynthesis of oxalic acid, the CDS of them are in close proximity to each other, even encoded on a single polycistronic message. The implication of this for our project is that we can make these two genes share the same promoter[2].


CAU-obcAB-gene.jpeg

Fig.3 Distribution of obcA and obcB[1].

We constructed a separate pET-obcB plasmid, and determined that the ObcB protein can be successfully expressed in E.coli BL21 (DE3) through SDS-PAGE, the results are as follows:

Sdsobcbpic.png

Fig.4 Obc protein in SDS-PAGE.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 669
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 669
    Illegal NotI site found at 582
    Illegal NotI site found at 687
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 650
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 669
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 669
    Illegal NgoMIV site found at 460
    Illegal NgoMIV site found at 501
    Illegal NgoMIV site found at 625
  • 1000
    COMPATIBLE WITH RFC[1000]



References

[1]Nakata, Paul A, and Cixin He. “Oxalic acid biosynthesis is encoded by an operon in Burkholderia glumae.” FEMS microbiology letters vol. 304,2 (2010): 177-82. doi:10.1111/j.1574-6968.2010.01895.x

[2]Oh, Juntaek et al. “Structural basis for bacterial quorum sensing-mediated oxalogenesis.” The Journal of biological chemistry vol. 289,16 (2014): 11465-11475. doi:10.1074/jbc.M113.543462