Detail of GM0023


Summary

The genetically modified enzybiotic, named PRF-119 , constructed by Domains Assembly strategy, improve the lytic activity.
With  PRF-119
  Sequence Length:  191 AA.
  Mass:  21480.9 Da.
  Isoelectric Point:  7.73
  Function:  the recombinant endolysin PRF-119 was highly active against all S. aureus isolates tested, including MRSA isolates, while CoNS isolates from clinical collections were not susceptible.

Construction

GM0023, constructed by Domains Assembly strategy.
    No schema construted on GM0023
Details:
GM0023[1-126]: derive from protein I6X5A8 sequence 26-160

I6X5A8, 495 AA., the LysK from Staphylococcus phage Fi200W

Source: Staphylococcus phage Fi200W

Domains and repeats

29-160:  IPR007921, the CHAP domain
196-369:  IPR002502, the N-acetylmuramoyl-L-alanine amidase domain
409-481:  IPR003646, the SH3-like domain, bacterial-type?

GO term prediction

Molecular Function:  0008745, the N-acetylmuramoyl-L-alanine amidase activity


Linking to UniprotKB
Linging to InterPro
GM0023[127-191]: 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 GM0023

Linking to UniprotKB
Linging to InterPro

Annotation

1-126
127-191

1-126
: EAD, CHAP domain, derived from endolysin of phageK;
127-191
: CBD, SH3b domain, derived from lysostaphin;
Domain and repeats:
127-187
: IPR003646,SH3-like domain, bacterial-type;
4-126
: IPR007921,CHAP domain;
GO term prediction:
SFGVMEAGAIDADGYYHAQCQDLITDYVLWLTDNKVRTWGNAKDQIKQSYGTGFKIHENKPSTVPKKGWIAVFTSGSYEQWGHIGIVYDGGNTSTFTILEQNWNGYANKKPTKRVDNYYGLTHFIE-YKSESASFTPNTDIITRTTGPFRSMPQSGVLKAGQTIHYDEVMKQDGHVWVGYTGNSGQRIYLPV
  BlastP to GMEnzy

Production

  1.  Expressed by pQE60 in E. coli XL1-Blue,  Purified by ion-exchange and size exclusion chromatography

Activity

  1.  MIC test on Staphylococcus aureus (398 methicillin-susceptible Staphylococcus aureus ) by microdilution broth method
  showed MIC50 0.098μg/ml,MIC90 0.391μg/ml
  2.  MIC test on Staphylococcus aureus (776 methicillin-resistant S. aureus isolates) by microdilution broth method
  showed MIC50 0.391μg/ml,MIC90 0.391μg/ml

Reference

  1. Idelevich, E. A.von Eiff, C.Friedrich, A. W.Iannelli, D.Xia, G.Peters, G.Peschel, A.Wanninger, I.Bec. (2011) In vitro activity against Staphylococcus aureus of a novel antimicrobial agent, PRF-119, a recombinant chimeric bacteriophage endolysin. Antimicrobial agents and chemotherapy. 09:4416-4419. [doi:10.1128/AAC.00217-11] [PMID:21746950] [FULL TEXT]

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