Detail of GM0035

Summary

The genetically modified enzybiotic, named LysK390 , constructed by Truncation strategy, decrease the lytic activity.
With  LysK390
  Sequence Length:  390 AA.
  Mass:  43365.6 Da.
  Isoelectric Point:  9.96
  Function:  show faint activity in both the zymogram and turbidity reduction assays, but no detectable activity in either plate lysis or MIC assays

Construction

GM0035, constructed by Truncation strategy.
No schema construted on GM0035

Details:

GM0035[1-390]: derive from protein I6X5A8 sequence 1-390

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

1-390

35-160

: EAD, CHAP domain, derived from staphylococcal bacteriophage endolysin LysK;

197-343

: EAD, Amidase domain, derived from staphylococcal bacteriophage endolysin LysK;

409-475

: CBD, SH3b domain, derived from staphylococcal bacteriophage endolysin LysK;
Domain and repeats:

196-369

: IPR002502,N-acetylmuramoyl-L-alanine amidase domain;

29-160

: IPR007921,CHAP domain;
GO term prediction:
0008745, the N-acetylmuramoyl-L-alanine amidase activity
0009253, the peptidoglycan catabolic process

MAKTQAEINKRLDAYAKGTVDSPYRVKKATSYDPSFGVMEAGAIDADGYYHAQCQDLITDYVLWLTDNKVRTWGNAKDQIKQSYGTGFKIHENKPSTVPKKGWIAVFTSGSYEQWGHIGIVYDGGNTSTFTILEQNWNGYANKKPTKRVDNYYGLTHFIEIPVKAGTTVKKETAKKSASKTPAPKKKATLKVSKNHINYTMDKRGKKPEGMVIHNDAGRSSGQQYENSLANAGYARYANGIAHYYGSEGYVWEAIDAKNQIAWHTGDGTGANSGNFRFAGIEVCQSMSASDAQFLKNEQAVFQFTAEKFKEWGLTPNRKTVRLHMEFVPTACPHRSMVLHTGFNPVTQGRPSQAIMNKLKDYFIKQIKNYMDKGTSSSTVVKDGKTSSAS

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 (23mM)/10μl
  2.  MIC test on Staphylococcus aureus (S. aureus Newman) by the turbidity-reduction assays
  showed 63±8ug/mL

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 GM0035

GMEnzy : A Genetically Modified Enzybiotic Database