Detail of GM0003


Summary

The genetically modified enzybiotic, named Lambda SA2-E-Lyso-SH3b , constructed by Domains Assembly strategy, change or extend lytic spectrum.
With  Lambda SA2-E-Lyso-SH3b
  Sequence Length:  208 AA.
  Mass:  22948.4 Da.
  Isoelectric Point:  6.67
  Function:  reduced the S. aureus bacterial load by 3 log units within 3 h

Construction

GM0003, constructed by Domains Assembly strategy.
C5WI88: Putative prophage LambdaSa2, lysin
P10547: Lysostaphin
UniProt

IPR008044:5-145
IPR013168:154-192
IPR013169:198-236
IPR002901:270-399
IPR003646:278-367
IPR003646:411-481
InterPro

GM0003:1-142
GM0003:143-205
GMEnzy

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390
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449
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493

Details:
GM0003[1-142]: derive from protein C5WI88 sequence 5-145

C5WI88, 449 AA., the Putative prophage LambdaSa2, lysin from Streptococcus dysgalactiae subsp. equisimilis (str

Source: Streptococcus dysgalactiae subsp. equisimilis (str

Domains and repeats

5-145:  IPR008044, the Bacteriophage lysin
6-138:  IPR000064, the Endopeptidase, NLPC/P60 domain
154-192:  IPR013168, the Cpl-7 lysozyme, C-terminal?
198-236:  IPR013169, the Cpl-7 lysozyme, C-terminal?
270-399:  IPR002901, the Mannosyl-glycoprotein endo-beta-N-acetylglucosamid

GO term prediction

Biological Process:  0009253, the peptidoglycan catabolic process
Molecular Function:  0004040, the amidase activity?


Linking to UniprotKB
Linging to InterPro
GM0003[143-205]: 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 GM0003

Linking to UniprotKB
Linging to InterPro

Annotation

1-142
143-205
50
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450
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550
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650
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942

1-142
: EAD, Endopeptidase domain, derived from streptococcal ??SA2 endolysin;
143-205
: CBD, SH3b domain, derived from lysostaphin;
Domain and repeats:
1-142
: IPR008044,Bacteriophage lysin;
143-201
: IPR003646,SH3-like domain, bacterial-type;
2-135
: IPR000064,Endopeptidase, NLPC/P60 domain;
GO term prediction:
TEKAISWMVARQGAVSYSMDYRNGPSSYDCSSAIYYALMSAGTISAGWAVNTEYMHDWLIKNGYVLIAENQDWNSQRGDVVIWGLRGQSAGAGGHVVMFVDSDNIIHCNYANNGITINNYNQTAASAGWMYSYVYRLATPATT-YKSESASFTPNTDIITRTTGPFRSMPQSGVLKAGQTIHYDEVMKQDGHVWVGYTGNSGQRIYLPV
  BlastP to GMEnzy

Production

  1.  Expressed by pET21a in E. coli BL21 (DE3),  Purified by immobilized metal ion affinity chromatography, using nickel-NTA Superflow resin(QIAGEN, Valencia, CA)

Activity

    No activity data found on GM0003

Reference

  1. Mathias Schmelcher, Anne M. Powell, Stephen C. Becker, Mary J. Camp, and David M. Donovan. (2012) Chimeric phage lysins act synergistically with Lysostaphin to kill mastitis causing Staphylococcus aureus in murine mammary glands. Appl. Environ. Microbiol.. 960:729-737. [doi:10.1128/AEM.07050-11] [PMID:22286996] [FULL TEXT]
  2. Stephen C. Becker, Juli Foster-Frey, Angeline J. Stodola, Daniel Anacker, David M. Donovan. (2009) Differentially conserved staphylococcal SH3b_5 cell wall binding domains confer increased staphylolytic and streptolytic activity to a streptococcal prophage endolysin domain. Gene. 4431-5:32-41. [doi:10.1016/j.gene.2009.04.023] [PMID:19422893]

Comments

  •   [1]  In a mouse model of mastitis, infusion of 25 μg of λSA2-E-Lyso-SH3b or λSA2-E-LysK-SH3b into mammary glands reduced S. aureus CFUs by 0.63 or 0.81 log units, compared to >2 log for lysostaphin.
  •   ----  ak0526@163.com   at  2013-09-06 16:07:47


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