Detail of GM0002


Summary

The genetically modified enzybiotic, named P128 , constructed by Domains Assembly strategy, change or extend lytic spectrum.
With  P128
  Sequence Length:  206 AA.
  Mass:  22757.7 Da.
  Isoelectric Point:  9.97
  Function:  The novel chimeric protein P128 is a staphylococcal cell wall-degrading enzyme under development for clearance of S. aureus nasal colonization and MRSA infection.

Construction

GM0002, constructed by Domains Assembly strategy.
Q6Y7R1: ORF56
P10547: Lysostaphin
UniProt

IPR007921:674-805
IPR003646:278-367
IPR003646:411-481
InterPro

GM0002:1-144
GM0002:145-208
GMEnzy

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493

Details:
GM0002[1-144]: derive from protein Q6Y7R1 sequence 669-808

Q6Y7R1, 808 AA., the ORF56 from Staphylococcus phage K

Source: Staphylococcus phage K

Domains and repeats

674-805:  IPR007921, the CHAP domain

GO term prediction

    No GO info found on GM0002

Linking to UniprotKB
Linging to InterPro
GM0002[145-208]: derive from protein P10547 sequence 411-485

P10547, 493 AA., the Lysostaphin from Staphylococcus simulans

Source: Staphylococcus simulans

Domains and repeats

239-382:  IPR011055, the Duplicated hybrid motif?
411-481:  IPR003646, the SH3-like domain, bacterial-type
278-367:  IPR003646, the Peptidase M23?

GO term prediction

    No GO info found on GM0002

Linking to UniprotKB
Linging to InterPro

Annotation

1-144
145-208
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1-144
: EAD, CHAP domain, derived from the phage tailassociated muralytic enzyme of Phage K;
145-208
: CBD, SH3b domain, derived from lysostaphin;
Domain and repeats:
145-204
: IPR003646,SH3-like domain, bacterial-type;
6-141
: IPR007921,CHAP domain;
GO term prediction:
MSLDSLKKYNGKLPKHDPSFVQPGNRHYKYQCTWYAYNRRGQLGIPVPLWGDAADWIGGAKGAGYGVGRTPKQGACVIWQRGVQGGSPQYGHVAFVEKVLDGGKKIFISEHNYATPNGYGTRTIDMSSAIGKNAQFIYDKK-YKSESASFTPNTDIITRTTGPFRSMPQSGVLKAGQTIHYDEVMKQDGHVWVGYTGNSGQRIYLPV
  BlastP to GMEnzy

Production

  1.  Expressed by inducible T7 expression system in E. coli ER2566,  Purified by 0 - 50% ammonium sulphate precipitation then anion exchange column

Activity

  1.  MIC test on Staphylococcus aureus (Staphylococcus aureus BK#13725) by the CLSI broth microdilution
  showed 2 μg/mL
  2.  MIC test on Staphylococcus aureus (Staphylococcus aureus BK#9894) by the CLSI broth microdilution
  showed 2 ug/mL

Reference

  1. Paul VD, Rajagopalan SS, Sundarrajan S, George SE, Asrani JY, Pillai R, Chikkamadaiah R, Durgaiah M,. (2011) A novel Bacteriophage Tail Associated Muralytic Enzyme (TAME) from Phage K and its development into a potent antistaphylococcal protein. BMC Microbiol. 110:226. [doi:10.1186/1471-2180-11-226.] [PMID:21985151] [FULL TEXT]
  2. Aradhana A Vipra,Srividya N Desai,Panchali Roy,Raghu Patil,Juliet M Raj,Nagalakshmi Narasimhaswamy,V. (2012) Antistaphylococcal activity of bacteriophage derived chimeric protein P128. BMC Microbiology. 2012:41. [doi:10.1186/1471-2180-12-41] [PMID:22439788] [FULL TEXT]

Comments

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