Detail of GM0033


Summary

The genetically modified enzybiotic, named LysK221 , constructed by Truncation strategy, decrease the lytic activity.
With  LysK221
  Sequence Length:  221 AA.
  Mass:  24649.8 Da.
  Isoelectric Point:  10.14
  Function:  show faint activity in both the zymogram and turbidity reduction assays, but no detectable activity in either plate lysis or MIC assays

Construction

GM0033, constructed by Truncation strategy.
    No schema construted on GM0033
Details:
GM0033[1-221]: derive from protein I6X5A8 sequence 1-221

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

Annotation

1-221

35-160
: EAD, CHAP domain, derived from staphylococcal bacteriophage endolysin LysK;
Domain and repeats:
29-160
: IPR007921,CHAP domain;
GO term prediction:
MAKTQAEINKRLDAYAKGTVDSPYRVKKATSYDPSFGVMEAGAIDADGYYHAQCQDLITDYVLWLTDNKVRTWGNAKDQIKQSYGTGFKIHENKPSTVPKKGWIAVFTSGSYEQWGHIGIVYDGGNTSTFTILEQNWNGYANKKPTKRVDNYYGLTHFIEIPVKAGTTVKKETAKKSASKTPAPKKKATLKVSKNHINYTMDKRGKKPEGMVIHNDAGRSS
  BlastP to GMEnzy

Production

  1.  Expressed by pET21a in E. coli BL21 (DE3),  Purified by nickel chromatography purification

Activity

  1.  MIC test on Staphylococcus aureus (Staphylococcus aureus strain Newman) by plate lysis assays and the turbidity-reduction assays
  showed 10 μg (39mM)/10μl

Reference

  1. Becker, S. C.Dong, S.Baker, J. R.Foster-Frey, J.Pritchard, D. G.Donovan, D. M.. (2009) LysK CHAP endopeptidase domain is required for lysis of live staphylococcal cells. FEMS microbiology letters. 01:52-60. [doi:10.1111/j.1574-6968.2009.01541.x] [PMID:19493008]

Comments

    No comments found on GM0033


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