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.
I6X5A8: LysK
P10547: Lysostaphin
UniProt

IPR007921:29-160
IPR002502:196-369
IPR003646:409-481
IPR003646:278-367
IPR003646:411-481
InterPro

GM0023:1-126
GM0023:127-191
GMEnzy

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495
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493

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
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988

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]

Comments

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