Coding

Part:BBa_K2488000

Designed by: Anastasiya Oguienko   Group: iGEM17_Moscow_RF   (2017-10-27)
Revision as of 22:29, 27 October 2017 by Sinicka tanua (Talk | contribs)

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Phytase of Obesumbacterium proteus

Phytases belong to the family of histidine acid phosphatases, a subclass of phosphatases capable of hydrolyzing phytates and releasing phosphate groups. A number of microorganisms, mainly of fungal and bacterial origin, are capable of phytase synthesis.Recently, artificial phytases with superior thermostability have been designed using semi-rational sequence comparison methods based upon several mesophilic homologues phytases from fungi (with sequences identity of about 50–70%). This phytase is naturally expressed in bacterium Obesumbacterium proteus. It has quite high thermostability and can deal with temperatures up to 60 degrees Celsius.

The phytase gene was isolated from Obesumbacterium proteus genomic library. The cleavage site of the PhyA signal peptide was predicted and experimentally proved. The PhyA protein shows maximum identity of 53% and 47% to phosphoanhydride phosphorylase from Yersinia pestis and phytase AppA from Escherichia coli, respectively. Based on protein sequence similarity of PhyA and its homologs, the phytases form a novel subclass of the histidine acid phosphatase family. To characterize properties of the PhyA protein, we expressed the phyA gene in E. coli. The specific activity of the purified recombinant PhyA was 310 U mg1 of protein. Recombinant PhyA showed activity at pH values from 1.5 through 6.5 with the optimum at 4.9. The temperature optimum was 40–45 C at pH 4.9. The Km value for sodium phytate was 0.34 mM with a Vmax of 435 U mg1. Zinin et al., FEMS Microbiology letters (2004).

For more details visit our wiki: [http://2017.igem.org/Team:Moscow_RF Team Moscow_RF]

And here is our new biobrick one of the main parts of which became phytase BBa_K248800: Obesumbacterium proteus phytase modified with dCBD-domain Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 4
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 56
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 319


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